Phosphoenolpyruvate Availability and the Biosynthesis of Shikimic

合集下载

邻苯二甲酸酯类的特性及在食品中的限量分析_黄婵媛

收稿日期：2013－11－20；修稿日期：2014－02－28作者简介：黄婵媛（1987－），女，硕士，研究方向为食品安全，通信地址：510110广东广州市越秀区八旗二马路38号前座301，E-mail ：hcy2011@gmail．com 。

邻苯二甲酸酯类的特性及在食品中的限量分析黄婵媛，蔡玮红，莫锡乾（广州市质量监督检测研究院，广州510110）摘要：邻苯二甲酸酯类（PAEs ）物质，作为塑料添加剂已有将近80年的历史，普遍存在于大气飘尘、工业废水、河流、土壤以及固体废弃物中，并已在食品、饮用水、人体体液中被检出，是一种全球最普遍的环境激素类污染物。

简要介绍了邻苯二甲酸酯类的特性，对国内外邻苯二甲酸酯类增塑剂在食品中的限量规定进行了分类和比较，客观分析了标准法规现状和存在的问题，并提出了建议。

关键词：邻苯二甲酸酯；特性；限量规定；标准法规中图分类号：TS201．6文献标志码：A 文章编号：1005－1295（2014）02－0066－04doi ：10．3969/j．issn．1005－1295．2014．02．017The Toxicity and Limited Provisions of Phthalate Esters in FoodHUANG Chan-yuan ，CAI Wei-hong ，MO Xi-qian（Guangzhou Quality Supervision and Testing Institute ，Guangzhou 510110，China ）Abstract ：Phthalate esters （PAEs ），as plastic additives ，have a history of nearly 80years．They are com-monly found in airborne particulates ，industrial wastewater ，rivers ，soil and solid waste ，and have been detected in food ，drinking water and body fluids．They are common worldwide environmental hormone pollutants．Ｒe-views on the characteristics of phthalates ，limited provisions and problems of domestic laws were elaborated and some suggestions were given．Key words ：Phthalate ester ；characteristic ；limited provision ；regulation0引言邻苯二甲酸酯类化合物是应用于塑料工业的主要增塑剂和软化剂，可以使塑料的柔韧性增强，容易加工，可用于工业用途［1］。

生物药剂学与药物动力学专业词汇

生物药剂学与药物动力学专业词汇※<A>Absolute bioavailability, F 绝对生物利用度Absorption 吸收Absorption pharmacokinetics 吸收动力学Absorption routes 吸收途径Absorption rate 吸收速率Absorption rate constant 吸收速率常数Absorptive epithelium 吸收上皮Accumulation 累积Accumulation factor 累积因子Accuracy 准确度Acetylation 乙酰化Acid glycoprotein 酸性糖蛋白Active transport 主动转运Atomic absorption spectrometry 原子吸收光谱法Additive 加和型Additive errors 加和型误差Adipose 脂肪Administration protocol 给药方案Administration route 给药途径Adverse reaction 不良反应Age differences 年龄差异Akaike’s information criterion, AIC AIC判据Albumin 白蛋白All-or-none response 全或无效应Amino acid conjugation 氨基酸结合Analog 类似物Analysis of variance, ANOVA ANOVA方差分析Anatomic Volume 解剖学体积Antagonism 拮抗作用Antiproliferation assays 抑制增殖法Apical membrane 顶端表面Apoprotein 载脂蛋白脱辅基蛋白Apparatus 仪器Apparent volume of distribution 表观分布容积Area under the curve, AUC 曲线下面积Aromatisation 芳构化Artery 动脉室Artifical biological membrane 人工生物膜Aryl 芳基Ascorbic acid 抗坏血酸维生素C Assistant in study design 辅助实验设计Average steady-state plasma drug concentration 平均稳态血浆药物浓度Azo reductase 含氮还原酶※<B>Backward elimination 逆向剔除Bacteria flora 菌丛Basal membrane 基底膜Base structural model 基础结构模型Basolateral membrane 侧底膜Bayesian estimation 贝易斯氏评估法Bayesian optimization 贝易斯优化法Bile 胆汁Billiary clearance 胆汁清除率Biliary excretion 胆汁排泄Binding 结合Binding site 结合部位Bioactivation 生物活化Bioavailability, BA 生物利用度Bioequivalence, BE 生物等效性Biological factors 生理因素Biological half life 生物半衰期Biological specimen 生物样品Biomembrane limit 膜限速型Biopharmaceutics 生物药剂学Bioequivalency criteria 生物等效性判断标准Biotransformation 生物转化Biowaiver 生物豁免Blood brain barrier, BBB BBB血脑屏障Blood clearance 血液清除率Blood flow rate-limited models 血流速度限速模型Blood flux in tissue 组织血流量Body fluid 体液Buccal absorption of drug 口腔用药的吸收Buccal mucosa 口腔粘膜颊粘膜Buccal spray formulation 口腔喷雾制剂※<C>Capacity limited 容量限制Carrier mediated transport 载体转运Catenary model 链状模型Caucasion 白种人Central compartment 中央室Characteristic 特点Chelate 螯合物Chinese Traditional medicine products 中药制剂Cholesterol esterase 胆固醇酯酶Chromatogram 色谱图Circulation 循环Classification 分类Clearance 清除率Clinical testing in first phase I期临床试验Clinical testing in second phase Ⅱ期临床试验Clinical testing in third phase Ⅲ期临床试验Clinical trial 临床试验Clinical trial simulation 临床实验计划仿真Clockwise hysteresis loop 顺时针滞后回线Collection 采集Combined administration 合并用药Combined errors 结合型误差Common liposomes, CL 普通脂质体Compartment models 隔室模型Compartments 隔室Competitive interaction 竞争性相互作用Complements 补体Complex 络合物Confidential interval 置信区间Conjugation with glucuronic acid 葡萄糖醛酸结合Controlled-release preparations 控释制剂Control stream 控制文件Conventional tablet 普通片Convergence 收敛Convolution 卷积Corresponding relationship 对应关系Corticosteroids 皮质甾体类Counter-clockwise hysteresis loop 逆时针滞后回线Countermeasure 对策Course in infusion period 滴注期间Covariance 协方差Covariates 相关因素Creatinine 肌酐Creatinine clearance 肌酐清除率Cytochrome P450, CYP450 细胞色素P450 Cytoplasm 细胞质Cytosis 胞饮作用Cytosol 胞浆胞液质※<D>Data File 数据文件Data Inspection 检视数据Deamination 脱氨基Deconvolution 反卷积Degree of fluctuation, DF DF波动度Delayed release preparations 迟释制剂Desaturation 降低饱和度Desmosome 桥粒Desulfuration 脱硫Detoxication 解毒Diagnosis 诊断Diffusion 扩散作用Dietary factors 食物因素Displacement 置换作用Disposition 处置Dissolution 溶解作用Distribution 分布Dosage adjustment 剂量调整Dosage form 剂型Dosage form design 剂型设计Dosage regimen 给药方案Dose 剂量dose-proportionality study 剂量均衡研究Dropping pills 滴丸Drug absorption via eyes 眼部用药物的吸收Drug binding 药物结合Drug concentration in plasma 血浆中药物浓度Drug Delivery System, DDS 药物给药系统Drug interaction 药物相互作用Drug-plasma protein binding ratio 药物—血浆蛋白结合率Drug-Protein Binding 药物蛋白结合Drug transport to foetus 胎内转运※<E>Efficient concentration range 有效浓度范围Efflux 外排Electrolyte 电解质Electro-spray ionization, ESI 电喷雾离子化Elimination 消除Elimination rate constant 消除速度常数Elongation 延长Emulsion 乳剂Endocytosis 入胞作用Endoplasmic reticulum 内质网Enterohepatic cycle 肠肝循环Enzyme 酶Enzyme induction 酶诱导Enzyme inhibition 酶抑制Enzyme-linked immunosorbent assays ELISA 酶联免疫法Enzymes or carrier-mediated system 酶或载体—传递系统Epithelium cell 上皮细胞Epoxide hydrolase 环化物水解酶Erosion 溶蚀Excretion 排泄Exocytosis 出胞作用Exons 外显子Experimental design 实验设计Experimental procedures 实验过程Exponential errors 指数型误差Exposure-response studies 疗效研究Extended least squares, ELS 扩展最小二乘法Extended-release preparations 缓控释制剂Extent of absorption 吸收程度External predictability 外延预见性Extraction ratio 抽取比Extract recovery rate 提取回收率Extrapolation 外推法Extravascular administration 血管外给药※<F>F test F检验Facilitated diffusion 促进扩散Factors of dosage forms 剂型因素Fasting 禁食Fibronectin 纤粘连蛋白First order rate 一级速度First Moment 一阶矩First order absorption 一级吸收First-order conditional estimation, FOCE 一级条件评估法First-order estimation, FO 一级评估法Fiest-order kinetics 一级动力学First pass effect 首过作用首过效应Fixed-effect parameters 固定效应参数Flavoprotein reductaseNADPH－细胞色素还原酶附属黄素蛋白还原酶Flow-through cell dissolution method 流室法Fluorescent detection method 荧光检测法Fraction of steady-state plasma drug concentration 达稳分数Free drug 游离药物Free drug concentration 游离药物浓度※<G>Gap junction 有隙结合Gas chromatography, GC 气相色谱法Gasrtointestinal tract, GI tract 胃肠道Gender differences 性别差异Generalized additive modeling, GAM 通用迭加模型化法Glimepiride 谷胱甘肽Global minimum 整体最小值Glomerular filtration 肾小球过滤Glomerular filtration rate, GFR 肾小球过滤率Glucuonide conjugation 葡萄糖醛酸结合Glutathione conjugation 谷胱甘肽结合Glycine conjugation 甘氨酸结合Glycocalyx 多糖—蛋白质复合体Goodness of Fit 拟合优度Graded response 梯度效应Graphic method 图解法Gut wall clearance肠壁清除率※<H>Half life 半衰期Health volunteers 健康志愿者Hemodialysis 血液透析Hepatic artery perfusion administration 肝动脉灌注给药Hepatic clearance, Clh 肝清除率Hierarchical Models 相同系列药物动力学模型High performance liquid chromatography, HPLC 高效液相色谱Higuchi equation Higuchi 方程Homologous 类似Human liver cytochrome P450 人类肝细胞色素P450 Hydrolysis 水解Hydroxylation 羟基化Hysteresis 滞后Hysteresis of plasma drug concentration 血药浓度滞后于药理效应Hysteresis of response 药理效应滞后于血药浓度※<I>Immunoradio metrec assays, IRMA 免疫放射定量法Incompatibility 配伍禁忌Independent 无关，独立Individual parameters 个体参数Individual variability 个体差异Individualization of drug dosage regimen 给药方案的个体化Inducer 诱导剂Induction 诱导Infusion 输注Inhibition 抑制Inhibitor 抑制剂Initial dose 速释部分Initial values 初始值Injection sites 注射部位Insulin 胰岛素Inter-compartmental clearance 隔室间清除率Inter-individual model 个体间模型Inter-individual random effects 个体间随机效应Inter-individual variability 个体间变异性Intermittence intravenous infusion 间歇静脉输液Internal predictability 内延预见性Inter-occasion random effects 实验间随机效应Intestinal bacterium flora 肠道菌丛Intestinal metabolism 肠道代谢Intra-individual model 个体内模型Intra-individual variability 个体内变异性Intramuscular administration 肌内给药Intramuscular injection 肌内注射Intra-peritoneal administration 腹腔给药Intravenous administration 静脉给药Intravenous infusion 静脉输液Intravenous injection 静脉注射Intrinsic clearance固有清除率内在清除率Inulin 菊粉In vitro experiments 体外试验In vitro–In vivo correlation, IVIVC 体外体内相关关系In vitro mean dissolution time, MDT vitro 体外平均溶出时间In vivo Mean dissolution time, MDT vivo 体内平均溶出时间Ion exchange 离子交换Isoform 异构体Isozyme 同工酶※<K>Kerckring 环状皱褶Kidney 肾※<L>Lag time 滞后时间Laplace transform 拉普拉斯变换Lateral intercellular fluid 侧细胞间隙液Lateral membrane 侧细胞膜Least detection amount 最小检测量Linearity 线性Linear models 线性模型Linear regression method 线性回归法Linear relationship 线性关系Lipoprotein 脂蛋白Liposomes 脂质体Liver flow 肝血流Local minimum 局部最小值Loading dose 负荷剂量Logarithmic models 对数模型Long circulation time liposomes 长循环脂质体Loo-Riegelman method Loo-Riegelman法Lowest detection concentration 最低检测浓度Lowest limit of quantitation 定量下限Lowest steady-state plasma drug concentration 最低稳态血药浓度Lung clearance 肺清除率Lymphatic circulation 淋巴循环Lymphatic system 淋巴系统※<M>Maintenance dose 维持剂量Mass balance study 质量平衡研究Masticatory mucosa 咀嚼粘膜Maximum likelihood 最大似然性Mean absolute prediction error, MAPE 平均绝对预测误差Mean absorption time, MAT 平均吸收时间Mean disintegration time, MDIT 平均崩解时间Mean dissolution time, MDT 平均溶出时间Mean residence time, MRT 平均驻留时间Mean sojourn time 平均逗留时间Mean squares 均方Mean transit time 平均转运时间Membrane-limited models 膜限速模型Membrane-mobile transport 膜动转运Membrane transport 膜转运Metabolism 代谢Metabolism enzymes 代谢酶Metabolism locations 代谢部位Metabolites 代谢物Metabolites clearance, Clm 代谢物清除率Method of residuals 残数法剩余法Methylation 甲基化Michaelis-Menten equation 米氏方程Michaelis-Menten constant 米氏常数Microbial assays 微生物检定法Microsomal P－450 mixed-function oxygenases 肝微粒体P-450混合功能氧化酶Microspheres 微球Microvilli 微绒毛Minimum drug concentration in plasma 血浆中最小药物浓度Mixed effects modeling 混合效应模型化Mixed-function oxidase, MFO 混合功能氧化酶Models 模型Modeling efficiency 模型效能Model validation 模型验证Modified release preparations 调释制剂Molecular mechanisms 分子机制Mono-exponential equation 单指数项公式Mono-oxygenase 单氧加合酶Mucous membrane injury 粘膜损伤Multi-compartment models 多室模型延迟分布模型Multi-exponential equation 多指数项公式Multifactor analysis of variance, multifactor ANOVA 多因素方差分析Multiple dosage 多剂量给药Multiple-dosage function 多剂量函数Multiple-dosage regimen 多剂量给药方案Multiple intravenous injection 多次静脉注射Myoglobin 肌血球素※<N>Naive average data, NAD 简单平均数据法Naive pool data, NPD 简单合并数据法Nanoparticles 纳米粒Nasal cavity 鼻腔Nasal mucosa 鼻粘膜National Institute of Health 美国国立卫生研究所Nephron 肾原Nephrotoxicity 肾毒性No hysteresis 无滞后Non-compartmental analysis, NCA 非隔室模型法Non-compartmental assistant Technology 非隔室辅助技术Nonionized form 非离子型Nonlinear mixed effects models, NONMEM 非线性混合效应模型Nonlinear pharmacokinetics 非线性药物动力学Non-linear relationship 非线性关系Nonparametric test 非参数检验※<O>Objective function, OF 目标函数Observed values 观测值One-compartment model 一室模型（单室模型）Onset 发生Open randomized two-way crossover design 开放随机两路交叉实验设计Open crossover randomized design 开放交叉随机设计Oral administration 口服给药Ordinary least squares, OLS 常规最小二乘法Organ 器官Organ clearance 器官清除率Original data 原始数据Osmosis 渗透压作用Outlier 偏离数据Outlier consideration 异常值的考虑Over-parameterized 过度参数化Oxidation 氧化Oxidation reactions 氧化反应※<P>Paracellular pathway 细胞旁路通道Parameters 参数Passive diffusion 被动扩散Pathways 途径Patient 病人Peak concentration 峰浓度Peak concentration of drug in plasma 血浆中药物峰浓度Poly-peptide 多肽Percent of absorption 吸收百分数Percent of fluctuation, PF 波动百分数Perfused liver 灌注肝脏Period 周期Peripheral compartments 外周室Peristalsis 蠕动Permeability of cell membrane 细胞膜的通透性P-glycoprotein, p-gp P-糖蛋白Phagocytosis 吞噬Pharmaceutical dosage form 药物剂型pharmaceutical equivalents 药剂等效性Pharmacokinetic models 药物动力学模型Pharmacokinetic physiological models 药物动力学的生理模型Pharmacological effects 药理效应Pharmacologic efficacy 药理效应Pharmacokinetics, PK 药物动力学Pharmacokinetic/pharmacodynamic link model 药物动力学-药效动力学统一模型Pharmacodynamics, PD 药效动力学Pharmacodynamic model 药效动力学模型Phase II metabolism 第II相代谢Phase I metabolism 第I相代谢pH-partition hypothesis pH分配假说Physiological function 生理功能Physiological compartment models 生理房室模型Physiological pharmacokinetic models 生理药物动力学模型Physiological pharmacokinetics 生理药物动力学模型Pigment 色素Physicochemical factors 理化因素Physicochemical property of drug 药物理化性质Physiological factors 生理因素Physiology 生理Physiological pharmacokinetic models 生理药物动力学模型Pinocytosis 吞噬Plasma drug concentration 血浆药物浓度Plasma drug concentration－time curve 血浆药物浓度-时间曲线Plasma drug-protein binding 血浆药物蛋白结合Plasma metabolite concentration 血浆代谢物浓度Plasma protein binding 血浆蛋白结合Plateau level 坪浓度Polymorphism 多态性Population average pharmacokinetic parameters 群体平均动力学参数Population model 群体模型Population parameters 群体参数Population pharmacokinetics 群体药物动力学Post-absorptive phase 吸收后相Post-distributive phase 分布后相Posterior probability 后发概率practical pharmacokinetic program 实用药代动力学计算程序Precision 精密度Preclinical 临床前的Prediction errors 预测偏差Prediction precision 预测精度Predicted values 拟合值Preliminary structural model 初始结构模型Primary active transport 原发性主动转运Principle of superposition 叠加原理Prior distribution 前置分布Prodrug 前体药物Proliferation assays 细胞增殖法Proportional 比例型Proportional errors 比例型误差Prosthehetic group 辅基Protein 蛋白质Pseudo-distribution equilibrium 伪分布平衡Pseudo steady state 伪稳态Pulmonary location 肺部Pulsatile drug delivery system 脉冲式释药系统※<Q、R>QQuality controlled samples 质控样品Quality control 质量控制Quick tissue 快分布组织RRadioimmuno assays, RIA 放射免疫法Random error model 随机误差模型Rapid intravenous injection 快速静脉注射Rate constants 速度常数Rate method 速度法Re-absorption 重吸收Receptor location 受体部位Recovery 回收率Rectal absorption 直肠吸收Rectal blood circulation 直肠部位的血液循环Rectal mucosa 直肠黏膜Reductase 还原酶Reduction 还原Reductive metabolism 还原代谢Reference individual 参比个体Reference product 参比制剂Relative bioavailability, Fr 相对生物利用度Release 释放Release medium 释放介质Release standard 释放度标准Renal 肾的Renal clearance, Clr 肾清除率Renal excretion 肾排泄Renal failure 肾衰Renal impairment 肾功能衰竭Renal tubular 肾小管Renal tubular re-absorption 肾小管重吸收Renal tubular secretion 肾小管分泌Repeatability 重现性Repeated one-point method 重复一点法Requirements 要求Research field 研究内容Reside 驻留Respiration 呼吸Respiration organ 呼吸器官Response 效应Residuals 残留误差Residual random effects 残留随机效应Reversal 恢复Rich Data 富集数据Ritschel one-point method Ritschel 一点法Rotating bottle method 转瓶法Rough surfaced endoplasmic reticulum 粗面内质网Routes of administration 给药途径※<S、T>SSafety and efficacy therapy 安全有效用药Saliva 唾液Scale up 外推Scale-Up/Post-Approval Changes, SUPAC 放大/审批后变化Second moment 二阶矩Secondary active transport 继发性主动转运Secretion 分泌Sensitivity 灵敏度Serum creatinine 血清肌酐Sigma curve 西格玛曲线Sigma-minus method 亏量法（总和减量法）Sigmoid curve S型曲线Sigmoid model Hill’s方程Simulated design 模拟设计Single-dose administration 单剂量（单次）给药Single dose response 单剂量效应Sink condition 漏槽条件Skin 皮肤Slow Tissue 慢分布组织Smooth surfaced endoplasmic reticulum 滑面内质网Soluble cell sap fraction 可溶性细胞液部分Solvent drag effect 溶媒牵引效应Stability 稳定性Steady-state volume of distribution 稳态分布容积Sparse data 稀疏数据Special dosage forms 特殊剂型Special populations 特殊人群Specialized mucosa 特性粘膜Species 种属Species differences 种属差异Specificity 特异性专属性Square sum of residual error 残差平方和Stagnant layer 不流动水层Standard curve 标准曲线Standard two stage, STS 标准两步法Statistical analysis 统计分析Statistical moments 统计矩Statistical moment theory 统计矩原理Steady state 稳态Steady state plasma drug concentration 稳态血药浓度Stealth liposomes, SL 隐形脂质体Steroid 类固醇Steroid-sulfatases 类固醇－硫酸酯酶Structure 结构Structure and function of GI epithelial cells 胃肠道上皮细胞的构造与功能Subcutaneous injections 皮下注射Subgroup 亚群体Subjects 受试者Sublingual administration 舌下给药Sublingual mucosa 舌下粘膜Subpopulation 亚群Substrate 底物Sulfate conjugation 硫酸盐结合Sulfation 硫酸结合Sum of squares 平方和Summation 相加Superposition method 叠加法Susceptible subject 易受影响的患者Sustained-release preparations 缓释制剂Sweating 出汗Synergism 协同作用Systemic clearance 全身清除率TTargeting 靶向化Taylor expansion 泰勒展开Tenous capsule 眼球囊Test product 试验制剂Therapy drug monitoring, TDM 治疗药物监测Therapeutic index 治疗指数Thermospray 热喷雾Three-compartment models 三室模型Though concentration 谷浓度Though concentration during steady state 稳态谷浓度Thromboxane 血栓素Tight junction 紧密结合Tissue 组织Tissue components 组织成分Tissue interstitial fluid 组织间隙Tolerance 耐受性Topping effect 尖峰效应Total clearance 总清除率Toxication and emergency treatment 中毒急救Transcellular pathway 经细胞转运通道Transdermal absorption 经皮肤吸收Transdermal drug delivery 经皮给药Transdermal penetration 经皮渗透Transport 转运Transport mechanism of drug 药物的转运机理Trapezoidal rule 梯形法Treatment 处理Trial Simulator 实验计划仿真器Trophoblastic epithelium 营养上皮层Two-compartment models 二室模型Two one sided tests 双单侧t检验Two period 双周期Two preparations 双制剂Two-way crossover bioequivalence studies 双周期交叉生物等效性研究Typical value 典型值※<U~Z>UUnwanted 非预期的Uniformity 均一性Unit impulse response 单位刺激反应Unit line 单位线Urinary drug concentration 尿药浓度Urinary excretion 尿排泄Urinary excretion rate 尿排泄速率VVagina 阴道Vaginal Mucosa 阴道黏膜Validation 校验Variance of mean residence time, VRT 平均驻留时间的方差Vein 静脉室Villi 绒毛Viscre 内脏Volumes of distribution 分布容积volunteers or patients studies 人体试验WWagner method Wagner法Wagner-Nelson method Wagner-Nelson法Waiver requirements 放弃（生物等效性研究）要求Washout period 洗净期Weibull distribution function Weibull分布函数Weighted Least Squares WLS加权最小二乘法Weighted residuals 加权残留误差XXenobiotic 外源物, 异生素ZZero Moment 零阶矩Zero-order absorption 零级吸收Zero-order kinetics 零级动力学Zero order rate 零级速度Zero-order release 零级释放。

Mycorrhizal manual of classical and molecular techniques

Mycorrhizal manual of classical and molecular techniques1.0 Manipulation and Staining of Spores and Roots 1.1. Spore and Root Extraction from Pot Cultures/Field Samples Remove soil sample from the rhizosphere of the host plant growing in the pot with a 10-20mm diameter core borer. If the sample is taken from the field larger quantities should be sieved (100g-200g) and mixed into a 1L beaker of water before pouring through the sieves. Clay based soils will block the finer sieve quickly and care must be taken to tap the base of that sieve to encourage excess water to drain through. The same procedure used for pot culture material should then be followed:a.Wash the soil through 710µm and 45µm pore sieves with running water.b.Remove root material trapped on the 710µm sieve to check forattached mycelium of AMF with spores or for staining of roots(Trypan blue, Chlorazole Black E, Alkaline Phosphatase, Acid Fuchsinetc.) if required.c.Backwash the contents of the 45µm sieve into a small beaker. Try tokeep the volume to a minimum.a.Swirl the beaker contents and quickly decant the contents into 50mlcentrifuge tubes up to a maximum half way up the tube.b.Gently inject an equal amount of a 60% (w/v) commercial sugar(sucrose) solution into the pellet at the bottom of each tube using asyringe with a plastic tube extension. There should be a clearinterface visible between the water (above) and sugar phase (below).c.Centrifuge the capped tubes at approx. 3000 rpm for 2 minutes in abench centrifuge.d.Remove the spores caught at the interface of the two layers with thesyringe and tube attachment. Start above the interface and workdown into the sugar phase using a circular motion as some speciesproduce spores which can sink in the sugar solution while others canfloat just above the interface.e.Pour the contents of the syringe into a clean 45µm sieve, and washthoroughly to remove traces of sugar solution.f.Backwash contents into a Petri dish and view under astereomicroscope1.2. Making a permanent slide mount for reference or BEG registrationa. After extracting spores from a fresh pot culture. Isolate a minimum of 10-20 spores.b. On two clean microscope slides place one drop each of the mountant PVLG (Polyvinyllactoglycerol) and Melzer's PVLG see annex 2.Transfer half the spores to the first drop of mountant and the second half to the second drop using fine tip forceps (e.g. VOMM forceps No. 999220: HWC 118-10 Hammacher Instruments, P. O. Box 120209, D-42677 Solingen, Germany)c. Try and orientate the spores so that distinguishing features will be apparent once the coverslip is added.d. Carefully place a clean coverslip over each drop, making sure to lower the coverslip at an angle to prevent air bubbles being trapped.e. Gently apply a pressure to the coverslips of one of the slides to break open the spores. Wait 30 seconds and then apply gentle pressure in a circular motion with a soft (B) pencil to break spore walls open further (The pressure will depend on the species of AMF). This should be done under a stereomicroscope.f. If using PVLG, remember to allow the mountant to polymerise and top-up it up as necessary before sealing with clear nail varnish or white/silver car paint.g. Label the slide at one end with the species name and reference code, date, your name, and the mountant used.1.3. Histochemical Staining of Total AMF Mycelium in RootsThe presence of arbuscular mycorrhizal fungi in roots is not visible without appropriate staining. Different non-vital strains are available (eg trypan blue, chlorazole black, fuschin) to detect intraradical mycelium and they enable an estimation of the abundance of arbuscular mycorrhizal fungi within a root system (Trouvelot et al, 1986). However, they stain both dead and living fungal structures.A fuller understanding of AM functioning requires consideration of the metabolic states of both internal and external hyphae, and the relationship between these, because the physiological interactions will necessitate the presence of an active symbiotic fungus. Activity of succinate dehydrogenase (SDH), a mitochondrial enzyme, is considered as an indicator of viability of mycorrhiza but does not appear to reflect mycorrhizal efficiency for plant growth enhancement (Vierheilig & Ocampo, 1989). Alkaline phosphatase (ALP) activity, located within the phosphate-accumulating vacuoles of AM hyphae (Gianinazzi et al., 1979) has been proposed as a physiological marker for analysing the efficiency of mycorrhiza (Tisserant et al., 1993). Measurements of these two enzyme activities make it easy to directly compare the total production of fungal tissue with the proportion that is living or functional, and to compare simultaneously the production of mycelium within roots and in soil in order to determine whether (i) biomass produced in the two compartiments is interdependent and, (ii) the proportion of metabolically active hyphae differs with time.1.3.1 Root Preparationa. Wash the roots free of soil.b. Cut roots into 1cm long segments.1.3.2 Trypan blue staining of total mycelium1.Clear roots in 2% (w/v) KOH (10% can be used for verypigmented tree roots) for 15 min at 120°C in a pressure cooker(1h at 90°C in a water bath or oven) (Do not use samples thatare more than 2g.2.Rinse roots with water three times on a fine sieve or using amesh and forceps.3.Cover roots with 2% (v/v) HCl for at least 30 mins andpreferably longer.4.Throw away the HCl and cover roots with 0.05% (w/v) trypanblue in lactoglycerol (1:1:1 lactic acid, glycerol and water 5:1:1may be used if tree roots are to be stained) for 15min at 120°Cin a pressure cooker or 15min to 1h at 90°C in water bath oroven.5.Place roo t s into Petri dish with 50% (v/v) glycerol fordestaining and viewing under stereomicroscope.Figure 3with the permission of Mark Brundett1.4 Histochemical staining of active AMF mycelium in roots1.4.1 Succinate dehydrogenase (SDH)- and alkaline phosphatase (ALP) staining of intraradical hyphae1.Wash roots from soil using ice cold water and keep in ice2.Cut roots into 1cm lengths and mix the roots sample uniform3.Take two 0.2-0.5g root samples4.Clear roots in the following solution 2h at room temperature :20ml 0.05 M Tris/citric acid pH 9.250mg/ml sorbitol15 units/ml cellulase (from A. niger)15 units/ml pectinase (from A. niger).5.Rinse roots with water on a fine sieve.6.Put the roots sample into two bottles marked with SDH and ALPseparately, and add 20ml solution A and B separately7.Incubate roots pieces overnight at room temperature8.Pour out mixture solution, wash with distilled water9.Put the roots marked SDH and ALP in sodium hypochloritesolution(containing 3% and 1% active chlorine separately) 5min,then wash with distilled water10.Transfer the roots into a Peri dish11.Observe purple-black or dark-brown particles in roots undermicroscope12.Estimate root length containing stained hyphae (see section 1.5and figure 3)Solution A for SDH stainingChemical Concentration Volume(ml)Tris/HCl (pH 7.4)0.2 mol.l-15MgCl2 5 mmol.l-12NBT 4 mg.ml-15H2O6Na-succinate 2.5 mol.l-12*NBT------ Nitro-blue Tetrazonium, prepared daily.Solution B for ALP stainingChemical Concentration Volume(ml)0.05 mol.l-118mlTris/citric acid (pH9.2)1 mg.ml-120mg-naphthyl acidphosphateFast Blue RR salt 1 mg.ml-120mgMgCl20.5 mg.ml-1 1 mlMnCl2.4H2O0.8 mg.ml-1 1 ml1.5. Estimation of AMF colonisationEstimation of mycorrhizal colonization according to Trouvelot et ala. Mount 15 root fragments on one slide; prepare two slides (30 root fragments total).b. Observe these fragments under the microscope and rate according to the range of classes indicated in figure 4 and Annex 1. These classes give a rapid estimation of the level of mycorrhizal colonisation of each root fragment and the abundance of arbuscules.c. Put the values into the computer program 'Mycocalc' to calculate the parameters: %F, %M, %m, %a and %A, according to Trouvelot et al.. 1986. (see Figure 4 from Trouvelot et al 1986)o Frequency of mycorrhiza in the root systemF% = ( nb of fragments myco/total nb)*100o Intensity of the mycorrhizal colonisation in the root system M% = (95n5+70n4+30n3+5n2+n1)/(nb total)where n5 = number of fragments rated 5; n4 = number offragments 4 etc.o Intensity of the mycorrhizal colonisation in the root fragments m% = M*(nb total)/(nb myco)o Arbuscule abundance in mycorrhizal parts of root fragments a% = (100mA3+50mA2+10mA1)/100where mA3, mA2, mA1 are the % of m, rated A3, A2, A1,respectively, withmA3=((95n5A3+70n4A3+30n3A3+5n2A3+n1A3)/nbmyco)*100/m and the same for A2 and A1.o Arbuscule abundance in the root systemA% = a*(M/100)Figure 41.6. Histochemical Staining of Total and Active Soil Mycelium1.6.1. Extraction and measurement of AM fungal hyphae in soil1.Take soil cores (1x6cm) randomly from pots.2.Mix the soil sample well and then put a 2 gram sample in a 500mlbeaker.3.Suspend the soil in 250ml distilled water.4.Filter the soil suspension through a 300 m mesh sieve.5.The washings are collected and blended 30 seconds at high speed in ablender.6.Transfer the suspension to a flask, shake by hand and then stand onthe bench for one minute.7.Pipette 10ml (5mlx2) aliquots onto a millipore filter(1.0m_m pore size)and filter under vacuum using the filter holder.8.Place the filter on a microscope slide and let dry9.Stain the hyphae on the filter in lactic glycerol-trypan blue (0.05%(v/v)) for 5 minutes.10.Observe the stained filter under a coverslip at 200X magnification.11.Examine 30 random fields and estimate hyphal length by using a gridline interception method as used for evaluating mycorrhizal rootlenthes (see Figure 5 from Brundett bet al 1996).Figure5with the permission of Mark Brundett1.6.2. Estimation of succinate dehydrogenase (SDH)- and alkaline phosphatase (ALP)- active hyphae in soil1.Take soil cores and put in a beaker on ice.2.Mix the soil sample well and take two 2 gram subsamples.3.Put the sub-samples in two bottles marked with SDH and ALPseparately, and cover the soil with 20ml ice-cold water immediately4.Add 20ml incubation solution A in the bottle marked with SDH, andadd 20ml solution B in the bottle marked with ALP.5.Incubate the soil suspension at room temperature for 3 hours.6.Filter the suspension through a 300 m mesh sieve with 210mldistilled water.7.The collected washings are blended at high speed for 25 minutes.8.Transfer the suspension to a flask and leave to stand on the bench forone minute.9.Pipette 10ml (5mlx2) aliquots on a millipore filter, filter under vacuumas above.10.Transfer the filter onto a microscope slide.11.Counterstain AMF hyphae on the filter with 0.1% basic fuschin for 5minutes.12.Cover the filter with a coverslip and observe under the microscope at200X magnification.13.Estimate stained hyphal length using the gridline intersect method.Solution A for SDH stainingChemical Concentration VolumeTris/HCl(pH7.4)0.2M5mlMgCl25mM2ml*NBT4mg/ml5mlH2O 2.5M6mlNa-succinate 2.5M2ml*NBT:Nitro-blue Tetrazonium,prepared daily.Solution B for ALP stainingChemical Concentration Volume0.05M18mlTris/citric acid (pH9.2)1mg/ml20mga-naphthyl acidphosphateFast Blue RR salt1mg/ml20mg10%MgCl20.5%1ml10%MnCl20.5%1ml1.7. References∙Boddington,C.L.; Bassett,E.E.; Jakobsen,I.; Dodd,J.C. 1999 Comparison of techniques for the extraction and quantification of extra-radical mycelium of arbuscular mycorrhizal fungi in soils. Soil Biol.Biochem. 31: (3)479-482.∙Brundrett, M., N. Bougher, B. Dell, T. Grove and N. Malajczuk 1996 Working with mycorrhizas in forestry and agriculture. ACIAR Monograph 32.374. ISBN 1 86320 181 5.∙Gianinazzi S, Gianinazzi-Pearson V & Dexheimer J (1979) Enzymatic studies on the metabolism of vesicular-arbuscular mycorrhiza. 3. Ultrastructural location of acid and alkaline phosphatase activity in onionroots infected by Glomus mosseae (Nicol. & Gerd.). New Phytol 82, 127-132.∙Tisserant B, Gianinazzi-Pearson V, Gianinazzi S & Gollotte A (1993) In planta histochemical staining of fungal alkaline phosphatase activity for analysis of efficient arbuscular mycorrhizal infections. Mycol. Res. 97,245-250.∙Trouvelot A, Kough JL & Gianinazzi-Pearson V (1986) Mesure du taux de mycorhization VA d’un système radiculaire. Recherche de méthodes d’estimation ayant une signification fonctionnelle. In : Physiological andGenetical Aspects of Mycorrhizae, V. Gianinazzi-Pearson and S. Gianinazzi (eds.). INRA Press, Paris, pp.217-221.∙Vierheilig H. & Ocampo JA (1989) Relationship between SDH-activity and VA mycorrhizal infection. Agriculture, Ecosystems and Environment 29, 439-442.see these website for further informations:BEG (Banque Européenne des Glomales) websiteMark Brundrett's Working with Mycorrhizas in Forestry and Agriculture2.0 - DNA Techniques: PCR of ribosomal DNA from spores2.1. Introduction to the Polymerase Chain ReactionThe Polymerase Chain Reaction (PCR) is an in vitro technique enabling chemical amplification of DNA. With the improvement brought by the use of the heat stable Taq DNA polymerase of Thermus aquaticus and automation it is possible to obtain quick amplification even of single copy genes, starting from minute amounts of material. The impact of this technique in molecular biology is comparable to that which followed the discovery of restriction enzymes. It has been adapted for a wide variety of applications, and in particular PCR has opened the possibility to analyse organisms at the nucleic acid level even when only small amounts of nucleic acid can be obtained, as in the case of arbuscular mycorrhizal (AM) fungi. Furthermore, although the efficiency of PCR amplification is dependent on the purity of the target DNA, Taq DNA polymerase is less sensitive to template purity than other molecular biology techniques so that partially purified nucleic acid can be used. This feature is a great advantage for plant/soil microbiology research, as investigations can be made directly on partially purified biological material, like fungal spores or infected plant roots.Ribosomal genes are multicopy genes tandemly organised in the genome. Each ribosomal genes encodes for three subunits (18S[SSU], 5.8S and 28S[LSU]) separated from each other by a Inter Non Transcribed region (ITS). The genes themselves are separated from each other by an Inter Genic Spacer (IGS) (see figure).The various characteristics of rRNA and rDNA have made them a choice target for phylogenetic and taxonomic studies, and comparative studies of the nucleotide sequences in ribosomal genes has provided data for the analysis of phylogenetic relationships over a wide taxonomic range of organisms. The nucleotidic polymorphism is not evenly distributed throughout the ribosomal genes and the three regions evolve at different rates. ITS and IGS are variable regions which mutate more frequently than the three conserved coding subunit regions (18S, 5.8S, 25S). This generally makes the former more informative for analyses of closely related genomes, whereas the coding regions of the small and the large ribosomal subunit are considered to be more useful for understanding more distant relationships at the species/order level.The internal transcribed spacer region like the intergenic spacer region, evolved much faster and sequence differences between different populations of one species, or in a single spore in the case of the Glomales, can be detected. The 5' end of the large ribosomal subunit harbours two informative polymorphic domains (D1 and D2). The polymorphism observed in these domains between and in a taxa, allows also to identify specific nucleotidic sequences which can be used to design primers with different level of specificity or discrimination (van Tuinen et al 1998a).2.2. Practical aspects of PCRThe Polymerase Chain Reaction is an in vitro technique which allows the amplification of a specific region of DNA located between two known sequences. After each cycle of denaturation, annealing and extension the amount of DNA is double. Potentially, after 20 cycles of PCR, there will be a 220- fold amplification (or 1.106). This illustrates the sensitivity of thismethod, and the potential artifactual amplification of DNA, as any traces of DNA can be amplified.SCHEMATIC REPRESENTATION OF THE POLYMERASE CHAINREACTIONBefore the discovery of thermostable polymerase, DNA polymerases such as the Klenow fragment of E. coli DNA polymerase I or T4 DNA polymerase were used. Due to their heat lability, fresh aliquots of enzymes had to be added after each denaturation cycle. The first heat stable DNA polymerase (Taq polymerase) was purified from Thermus aquaticus . Today several heat stable polymerase are available, they are of natural or recombinant origin and vary in their biochemical properties such as extension rate, thermal stability, 5'?3' or 3'?5' exonuclease activity. The specificity and activity of the same enzymes is also very dependent on the producer. Some enzymes such as Tth, have a reverse transcriptase activity, they cannot therefore be used for the synthesis of cDNA.Beside the enzyme the other factors that can affect the PCR reaction are:∙Primers∙M gCl2 concentration∙Primer concentration∙Primer sequence∙Reaction stringency∙Length of the amplification product∙Number of PCR cycles∙other unknown factorsFor each PCR reaction the optimal conditions can vary depending mainly on the primer-DNA combination.The dNTP's are generally used at a concentration of 100µM, although at lower concentrations (10-100 µM) Taq polymerase has a higher fidelity. The most common buffer used with the Taq polymerase is:∙10 mM Tris/HCl pH 8,3∙50 mM KCl∙ 1.5 mM MgCl2∙0.1% (w/v) gelatinThe MgCl2 concentration affects the specificity of the PCR reaction. A too low concentration affects the final yield whereas a too high concentration reduces the specificity of the reaction. Other components often present in DNA extraction buffer can affect the enzyme activity. SDS at a concentration > 0.01% inhibits the polymerase. The inhibition of SDS (0.01%) can be reversed by some non-ionic detergents (0.5 % (v/v) Tween 20, NP 40). The primer working concentration is generally of 0.5 - 1 µM. If the primer concentration is too high primer dimerisation can occur.The primer composition is very important. In most PCR applications, the primers are designed to be exactly complementary to the template DNA. The general rules for the primer design are: a length of about 20 - 30 nucleotides. Shorter primers can be used with success and primers longer than 30 do not increase the specificity of the bindingthe GC content should be about 50%the 3' ends should not be complementary, as primer dimerisation will occur the 3' of the primer should be as homologous as possiblethe 5' can be modified to add a restriction site or a GC clamp, in this case, both primers should be equivalent in their melting temperaturesThe number of the cycles can be increased to increase the amount of product recovered, but this will also increase non-specific amplification. Beside all these factors, some primer combinations will work very well, and others not. As so many factors affect the PCR reaction it is very important to have a positive and negative control in an PCR reaction.2.3. ContaminationAs the PCR reaction is so sensitive, precautions have to be taken to avoid undesirable amplifications., such as using DNA free water and negative controls with every set of amplifications.Thermostable DNA polymerases and their sourcesDNA polymerase Natural/recombinant SourceTaq Natural Thermus aquaticusAmplitaq®Recombinant T. aquaticusAmplitaq® (StoffelRecombinant T. aquaticusfragment)Hot Tub TM Natural Thermus flavisPyrostase TM Natural T. flavisVent TM Recombinant Thermoccucus litoralisDeepVent TM Recombinant Pyrococcus GB-DTth Recombinant Thermus thermophilusPfu Natural Pyrococcus furiosusPfu Cloned Pyrococcus furiosuExo-PFU Recombinat Pyrococcus furiosuUITma TM Recombinant Thermotoga maritimaProperties of DNA polymerases commonly used in PCRTaq/ Amplitaq®StoffelfragmentVent TM Deep-VentTMPfu Tth UITma TMThermostability-half-life at 95°C40804001380>12020>505’ - 3’exonucleaseactivityYes No No No No Yes No3’ - 5’exonucleaseactivityNo No Yes Yes Yes No YesExtension rate(nt/sec)75>50>80n.i.n.i.30-40n.i.ReversetranscriptaseactivityWeak Weak n.i.n.i.n.i.Yes n.i.Resulting DNAends3’A3’A>95%blunt >95%bluntn.i.3’A BluntMolecular weight(kDa)9461n.i.n.i.929470from : PCR Newton, C.R. and Graham, A. BIOS Scientific Publishers Limited 19942.4. PCR from AMFWe present a protocol which has been used to amplify the 5' end of the large ribosomal unit of Glomales, using the fungal spore as starting material.This method can be applied to other types of biological material, like plant roots (van Tuinen et al 1998b; Jacquot et al. 2000; Turnau et al. 2001)2.4.1. Preparation of the nucleic acidsa. Collect clean and shiny Glomalean spores (1 to 10) with forceps under a binocular microscope and rinse with distilled water.b. Transfer the spores to a 1.5 ml Eppendorf tube containing 10 µl water and crush by means of a micropestle, or a glass Pasteur pipette. Disposable micropestles are available from many laboratory suppliers, and can be reused after incubation for several hours in 0.1 N NaOH to digest any remaining DNA.c. Add 30µl 100 mM Tris/HCl pH 8.0 and 10 µl of 20% Chelex 100 (Bio Rad) to the crushed spores. Vortex this suspension and then bring to 95 °C for 5 min. Cool on ice.d. Clear the suspension by centrifugation for 1 min and discard the pellet. The supernatant contains the nucleic acids for the PCR reactions. Depending on the nature of the species analysed, and especially its DNA content, the supernatant obtained can be directly used as template for PCR amplification, or be diluted up to 1/100 before use. This DNA preparation should stored at -20 °C until use.2.4.2. Preparation of the PCR reaction1.Each PCR reaction is performed in a final volume of 50 µl.2.For each set a amplification reaction a negative control, without DNA,has to be made.∙Each reaction mixture contains:For convenience and to minimise the risk of contamination, a master mixture is prepared with all the reagents except the template DNA.∙For PCR machine without heated lid, 25 - 50 µl mineral oil is laid over the mixture, and quickly spun down onto the surface.∙25 to 35 PCR cycles are performed as follows:∙Denaturation: at 95 °C for 3 minutes for the initial denaturation,93 °C and 45 seconds for the remaining cycles.∙Annealing: at 62 °C for 45 seconds.∙Extension: at 72 °C for 45 seconds (1 minute per 1 kb is an average polymerisation speed of Taq DNA polymerase).∙ A final extension of 5 minutes is performed at the end of the cycles.2.4.3. Gel electrophoresisElectrophoresis through a medium such as agarose or polyacrylamide is a standard method for the separation and purification of nucleic acids. As nucleic acids are charged molecules they will migrate when exposed to an electric field. The size of the molecules to be resolved will, influence the choice of the electrophoretic separation media. For fragments up to 500 bp, polyacrylamide gels are the most effective. Whereas for larger molecules agarose will be the medium of choice. Similar to polyacrylamide gel electrophoresis, there is a linear relationship between agarose concentration and the logarithm of the molecular weight of the DNA. Range of Separation In Gels Containing Different Amounts of AgaroseAmounts of agarose in gel in TAE (% [w/v])Efficient range of separation of linear DNA molecules (kb)0.35-600.61-200.70.8-100.90.5-71.20.4-61.50.2-32.00.1-2The migration of the DNA molecule also depends on it's conformation. The DNA molecule can be superhelical (form I), nicked circular (form II) or linear (form III). Depending on the electrophoretic conditions (ionicstrength of the buffer, intensity of the electric field) the form I can migrate faster then the linear form.Generally the DNA molecule is visualised after electrophoresis by staining with ethidium bromide (EtBr).BE CAREFUL WHEN MANIPULATING Ethidium Bromide.IT IS A POWERFUL MUTAGEN.Ethidium bromide is a fluorescent dye which intercalates between the bases of DNA. After irradiation with UV light the bound dye retransmits the light at 590 nm. Through this staining, which can be done during or after the electrophoresis, small amounts of DNA (<10 ng) can be detected.2.4.4. Nested PCR ReactionThe aim of the nested PCR reaction is to increase the specificity of the amplification reaction by performing two PCR amplifications one after the other.The first PCR reaction is performed as previously described, but for the second reaction the amplification products obtained in the first amplification cycles are used as template, after a dilution of up to 103, an internal primer.In this way the specificity of the amplification is increased as the target DNA to be amplified requires to possess the three primer binding, the efficiency of the amplification is increased as the number of cycles can be increased, without loss of specificity.Protocola. After the first PCR amplification, the reaction is checked by loading 5 µl of the amplification product on an 1.2 % agarose gel.b. For the nested PCR reaction 5µl of the amplification product diluted 500x, are used as target for the second round of amplification (25 cycles)c. The annealing temperature will depend on the primer pair used.Abbreviations:SDS: sodium dodecyl sulfatedNTP: deoxynucleosides triphosphateTAE: Tris-acetate (40 mM Tris-acetate pH 8.0 ; 1 mM EDTA)TE: Tris-EDTA (10 mM Tris/HCl pH 7.4-8.0 ; 1 mM EDTA)EDTA: ethylenediaminetetraacetate2.5. References∙Jacquot-Plumey, E van Tuinen, D Gianinazzi, S and Gianinazzi-Pearson, V 2000 Monitoring species of arbuscular mycorrhizal fungi in planta and in soil by nested PCR : application to the study of the impact of sewage sludge.Plant Soil 226, 179-188.∙Turnau, K Ryszka, P Gianinazzi-Pearson, V and van Tuinen, D 2001 Identification of arbuscular mycorrhizal fungi in soils and roots of plants colonizing zinc wastes in southern Poland. Mycorrhiza 10, 169-174.∙van Tuinen, D Zhao, B and Gianinazzi-Pearson, V 1998a PCR in studies of AM Fungi: from Primers to Application.In Mycorrhiza Manual . A. K. Varma (eds) Springer-Verlag: Heidelberg , pp. 387-400∙van Tuinen, D Jacquot, E Zhao, B Golotte, A and Gianinazzi-Pearson, V 1998b Characterization of root colonization profiles by a microcosm community of arbuscular mycorrhizal fungi using 25S rDNA-targetednested PCR. Mol Ecol 7, 103-111.3.0 DNA Techniques: PCR-SSCP analysis3.1. IntroductionThe ultimate character that can be used to distinguish species is variation in DNA sequence between homologous genes or regions. The distinguishing。

文档：色谱法英文

“固定相分子筛载气归一化法微量进样器进样保留时间峰面积The stationary phase is the substance which is fixed in place for the chromatography procedure.A molecular sieve is a material containing tiny pores of a precise and uniform size that is used as an adsorbent for gases and liquids.When the mobile phase is gas, it is called eluant gas.The mobile phase is the phase which moves in a definite direction. In statistics, normalization refers to the division of multiple sets of data by a common variable in order to negate that variable's effect on the data, thus allowing underlying characteristics of the data sets to be compared.Microinjector is a kind of injector which can make injection in a very small volumn. An injector, ejector, steam ejector or steam injector is a pump-like device that uses the Venturi effect of a converging-diverging nozzle to convert the pressure energy of a motive fluid to velocity energy which creates a low pressure zone that draws in and entrains a suction fluid and then recompresses the mixed fluids by converting velocity energy back into pressure energy.Injection is a method of putting sample into the column with a syringe or injectorThe retention time is the characteristic time it takes for a particular analyte to pass through the system (from the column inlet to the detector) under set conditions.The area of a peak is called peak area.色谱图 chromatogram色谱峰 chromatographic peak峰底 peak base峰高 h，peak height峰宽 W，peak width半高峰宽 Wh/2，peak width at half height峰面积 A，peak area拖尾峰 tailing area前伸峰 leading area假峰 ghost peak畸峰 distorted peak反峰 negative peak拐点 inflection point原点 origin斑点 spot区带 zone复班 multiple spot区带脱尾 zone tailing基线 base line基线漂移 baseline drift基线噪声 N，baseline noise统计矩 moment一阶原点矩γ1，first origin moment二阶中心矩μ2，second central moment三阶中心矩μ3，third central moment液相色谱法 liquid chromatography，LC液液色谱法 liquid liquid chromatography，LLC液固色谱法 liquid solid chromatography，LSC正相液相色谱法 normal phase liquidchromatography反相液相色谱法 reversed phase liquidchromatography，RPLC柱液相色谱法 liquid column chromatography高效液相色谱法 high performance liquidchromatography，HPLC尺寸排除色谱法 size exclusion chromatography，SEC凝胶过滤色谱法 gel filtration chromatography凝胶渗透色谱法 gel permeation chromatography，GPC亲和色谱法 affinity chromatography离子交换色谱法 ion exchange chromatography，IEC离子色谱法 ion chromatography离子抑制色谱法 ion suppression chromatography离子对色谱法 ion pair chromatography疏水作用色谱法 hydrophobic interactionchromatography制备液相色谱法 preparative liquid chromatography平面色谱法 planar chromatography纸色谱法 paper chromatography薄层色谱法 thin layer chromatography，TLC高效薄层色谱法 high performance thin layerchromatography，HPTLC 浸渍薄层色谱法 impregnated thin layerchromatography凝胶薄层色谱法 gel thin layer chromatography离子交换薄层色谱法 ion exchange thin layerchromatography制备薄层色谱法 preparative thin layerchromatography薄层棒色谱法 thin layer rod chromatography液相色谱仪 liquid chromatograph制备液相色谱仪 preparative liquid chromatograph凝胶渗透色谱仪 gel permeation chromatograph涂布器 spreader点样器 sample applicator色谱柱 chromatographic column棒状色谱柱 monolith column monolith column微粒柱 microparticle column填充毛细管柱 packed capillary column空心柱 open tubular column微径柱 microbore column混合柱 mixed column组合柱 coupled column预柱 precolumn保护柱 guard column预饱和柱 presaturation column浓缩柱 concentrating column抑制柱 suppression column薄层板 thin layer plate浓缩区薄层板 concentrating thin layer plate荧光薄层板 fluorescence thin layer plate反相薄层板 reversed phase thin layer plate梯度薄层板 gradient thin layer plate烧结板 sintered plate展开室 development chamber往复泵 reciprocating pump注射泵 syringe pump气动泵 pneumatic pump蠕动泵 peristaltic pump检测器 detector微分检测器 differential detector积分检测器 integral detector总体性能检测器 bulk property detector溶质性能检测器 solute property detector(示差)折光率检测器 [differential] refractive indexdetector 荧光检测器 fluorescence detector紫外可见光检测器 ultraviolet visible detector电化学检测器 electrochemical detector蒸发(激光)光散射检测器 [laser] light scatteringdetector光密度计 densitometer薄层扫描仪 thin layer scanner柱后反应器 post-column reactor体积标记器 volume marker记录器 recorder积分仪 integrator馏分收集器 fraction collector工作站 work station固定相 stationary phase固定液 stationary liquid载体 support柱填充剂 column packing化学键合相填充剂 chemically bonded phasepacking薄壳型填充剂 pellicular packing多孔型填充剂 porous packing吸附剂 adsorbent离子交换剂 ion exchanger基体 matrix载板 support plate粘合剂 binder流动相 mobile phase洗脱(淋洗)剂 eluant，eluent展开剂 developer等水容剂 isohydric solvent改性剂 modifier显色剂 color [developing] agent死时间 t0，dead time保留时间 tR，retention time调整保留时间 t'R，adjusted retention time死体积 V0，dead volume保留体积 vR，retention volume调整保留体积 v'R，adjusted retention volume柱外体积 Vext，extra-column volune粒间体积 V0，interstitial volume(多孔填充剂的)孔体积 VP，pore volume of porouspacking 液相总体积 Vtol，total liquid volume洗脱体积 ve，elution volume流体力学体积 vh，hydrodynamic volume相对保留值 ri.s，relative retention value分离因子α，separation factor流动相迁移距离 dm，mobile phase migrationdistance流动相前沿 mobile phase front溶质迁移距离 ds，solute migration distance比移值 Rf，Rf value高比移值 hRf，high Rf value相对比移值 Ri.s，relative Rf value保留常数值 Rm，Rm value板效能 plate efficiency折合板高 hr，reduced plate height分离度 R，resolution液相载荷量 liquid phase loading离子交换容量 ion exchange capacity负载容量 loading capacity渗透极限 permeability limit排除极限 Vh，max，exclusion limit拖尾因子 T，tailing factor柱外效应 extra-column effect管壁效应 wall effect间隔臂效应 spacer arm effect边缘效应 edge effect斑点定位法 localization of spot放射自显影法 autoradiography原位定量 in situ quantitation生物自显影法 bioautography归一法 normalization method内标法 internal standard method外标法 external standard method叠加法 addition method普适校准(曲线、函数) calibration function or curve谱带扩展(加宽) band broadening(分离作用的)校准函数或校准曲线 universalcalibration function or curve [of separation] 加宽校正 broadening correction加宽校正因子 broadening correction factor溶剂强度参数ε0，solvent strength parameter洗脱序列 eluotropic series洗脱(淋洗) elution等度洗脱 gradient elution梯度洗脱 gradient elution(再)循环洗脱 recycling elution线性溶剂强度洗脱 linear solvent strength gradient程序溶剂 programmed solvent程序压力 programmed pressure程序流速 programmed flow展开 development上行展开 ascending development下行展开 descending development双向展开 two dimensional development环形展开 circular development离心展开 centrifugal development向心展开 centripetal development径向展开 radial development多次展开 multiple development分步展开 stepwise development连续展开 continuous development梯度展开 gradient development匀浆填充 slurry packing停流进样 stop-flow injection阀进样 valve injection柱上富集 on-column enrichment流出液 eluate柱上检测 on-column detection柱寿命 column life柱流失 column bleeding显谱 visualization活化 activation反冲 back flushing脱气 degassing 沟流 channeling 过载 overloading。

Use of the Yeast Two-Hybrid System to Identify effector

C hapter 11 U se of the Yeast Two-Hybrid System to IdentifyTargets of Fungal EffectorsShunwen LuAbstractT he yeast two-hybrid (Y2H) system is a binary method widely used to determine direct interactions between paired proteins. Although having certain limitations, this method has become one of the two main systemic tools (along with afﬁ nity puriﬁ cation/mass spectrometry) for interactome mapping in model organisms including yeast, A rabidopsis,and humans. It has also become the method of choice for investi-gating host–pathogen interactions in fungal pathosystems involving crop plants. This chapter describes general procedures to use the GAL4-based Y2H system for identiﬁ cation of host proteins that directly interact with proteinaceous fungal effectors, thus being their potential targets. The procedures described include cDNA library construction through in vivo recombination, library screening by yeast mating and cotransformation, as well as methods to analyze positive clones obtained from library screening. These procedures can also be adapted to conﬁ rmation of suspected interactions between characterized host and pathogen proteins or determination of interacting domains in partner proteins.K ey words:F ungal pathogens ,P lant disease ,P rotein–protein interactions ,Y east transformation , S ynthetic dropout media ,R eporter genes ,A utoactivation ,A ureobasidin A ,a-Galactosidase assay 1.IntroductionT he yeast two-hybrid (Y2H) system was ﬁ rst pioneered by Fieldsand Song in 1989 (1)and has been since widely used to studyprotein–protein interactions in various binary systems (2–7)and,more recently, to establish interactomes in model organisms includingyeast, A rabidopsis,and humans (8–11). This system was originallydeveloped based on the intriguing properties of the GAL4 proteinof the baker’s yeast S accharomyces cerevisiae. GAL4 is a mastertranscription activator controlling expression of several G ALgenes encoding enzymes involved in galactose utilization (12). Melvin D. Bolton and Bart P.H.J. Thomma (eds.), Plant Fungal Pathogens: Methods and Protocols,Methods in Molecular Biology,vol. 835, DOI 10.1007/978-1-61779-501-5_11, © Springer Science+Business Media, LLC 2012165166S. LuGAL4 consists of two separable and functionally essential domains:an N-terminal DNA-binding domain (BD) which binds to speciﬁ cDNA sequences called upstream-activating sequences (UAS), anda C-terminal activation domain (AD) which interacts with basal tran-scriptional machinery (Fig. 1). In Y2H assays, the two testing proteins(the “bait” and the “prey”) are fused in frame with the BD and ADdomains, respectively, and coexpressed in a yeast strain which carriestwo or more reporter genes. A physical interaction between thebait and prey proteins would reconstitute proximity of the nativeGAL4 domains, thus activating the reporter genes (Fig. 1). Apartfrom testing the interaction between two known proteins as originallydesigned (1), Y2H can be also used for library screening to identifythe interacting partner(s) of a characterized protein (the “bait”)without prior knowledge of the nature of the “prey.” This applicationhas been adapted to identify novel targets of many functionallycharacterized proteins (8–11, 13)including proteinaceous fungaleffectors involved in plant pathogenesis (14, 15).P roteinaceous fungal effectors, e.g., avirulence (Avr) proteinsand certain host-selective toxins (HSTs), are of diverse proteins(usually small and cysteine-rich) secreted by the fungal pathogenand recognized by the host plant, in particular host–pathogeninteractions (16–18). Although speciﬁc interactions between thecharacterized fungal effectors and the corresponding host resistance(or susceptibility) genes may have been well characterized, thecognate host receptors or targets of most fungal effectors arestill unknown. It is often difﬁ cult to assign candidate interactingpartners for fungal effector proteins simply by exploring availableinteractomes or comparative proteomics databases because mostfungal effectors have no apparent homologies to any known proteinsMPUAS reporter geneADE2, HIS3, URA3MEL1, lacZAUR1-CF ig. 1. S chematic diagram showing the principle of the GAL4-based yeast two-hybrid system(adapted from Fields and Song (1)). A physical interaction between the bait and prey fusionproteins brings the GAL4 domains into close proximity and activates the reporter gene.U AS upstream-activating sequence; M P minimal promoter. Enzyme activities encoded bythe reporter genes: Nutritional: A DE2phosphoribosylaminoimidazole carboxylase(adenine biosynthesis); H IS3imidazoleglycerol-phosphate dehydratase (histidine biosyn-thesis); U RA3orotidine-5¢-phosphate decarboxylase (uracil biosynthesis). Colormetric:M EL1secreted a-galactosidase;lacZ intracellular b-galactosidase.Antibiotic resistance:A UR-1C mutated phosphatidylinositol: ceramide phosphoinositol transferase (conferringresistance to aureobasidin A). The speciﬁc reporter genes in related yeast strains areindicated in Table 1.16711 Use of the Yeast Two-Hybrid System to Identify Targets of Fungal Effectors( 17, 18 ) . ToxA, the major proteinaceous HST produced by the wheattan spot fungus P yrenophora tritici-repentis ( 19, 20 ) and the leaf/glume blotch fungus S tagonospora nodorum ( 21) , has been shown to interact with two chloroplast proteins, ToxABP1 (a homologue of A rabidopsis THYLAKOID FORMATION 1 protein) ( 14 ) and PCN(a plastocyanin) (15 ) . However, the host speciﬁ city-determining target(s) of ToxA is still unknown because in both ToxA-sensitive and -insensitive wheat lines, T oxABP1 and P CN are found to be identical (100% nucleotide identity) and expressed in the samepatterns ( 14,15 ) . Furthermore, ToxA does not interact directly with T sn1 , a disease resistance-like gene which governs the ToxA-triggered susceptibility in wheat ( 22) . These ﬁ ndings emphasize the complexity of effector–host interactions and the need for an efﬁ cient method to unravel the interacting networks.T his chapter is intended to describe general experimental procedures to use the GAL4-based Y2H system for identiﬁ cation of host plant proteins that directly interact with proteinaceous fun-gal effectors, thus being their potential targets. These procedures include generation of the bait construct and expression strain, cDNA library construction through in vivo recombination, library screening by yeast mating and cotransformation, as well as analysis of the library-derived positive clones. The methods described have been used successfully to identify two pathogenesis-related proteins that are differentially expressed in ToxA-sensitive and -insensitive wheat lines and interact directly with ToxA (Lu, S., Friesen, T.L., Faris, J.D., unpublished). In addition, the materials and methods used for library screening can also be adapted for conﬁ rmation of suspected interactions between characterized host and pathogen proteins or determination of interacting domains in identiﬁ ed protein partners. This chapter is not intended to give a comprehensive review on the rapidly evolving Y2H systems and the related protocols pub-lished during the last decades, but some key references are cited in related sections. A brief introduction on biochemical methods that can be used for further validation of physical interactions between paired proteins is given in the last section of the chapter.Plasmids and yeast strains are described using examples from the MatchMaker Y2H system (Table1 ).P lasmids used in the Y2H system act as shuttle vectors that can be maintained in E . coli and in S . cerevisiae . pGBKT7 (Fig.2a ) and pGADT7 (Fig.2b ) are the two common vectors for construction of the bait and the prey fusion proteins, respectively. pGADT7-Rec(Fig.2b ) is a prey library vector which contains a multiple cloning 2.Materials 2.1.Plasmidsand Yeast Strains2.1.1.Plasmids168S. LuT a b l e 1G A L 4-b a s e d y e a s t t w o -h y b r i d l i b r a r y s c r e e n i n g s y s t e m s aC o m p a n y Y 2H s y s t e m b f e a t u r e sG A L 4-B D p l a s m i d c s e l e c t i o n m a r k e rG A L 4-A D p l a s m i d ds e l e c t i o n m a r k e rY e a s t s t r a i n r e p o r t e r g e n e s aT h i s t a b l e i s g i v e n t o s h o w t h e a v a i l a b i l i t y o f m a j o r G A L 4-b a s e d Y 2H s y s t e m s o n l y a n d i s n o t i n t e n d e d t o r e c o m m e n d a s p e c i ﬁ c c o m m e r c i a l s o u r c eb D e t a i l e d p r o d uc t i n f o r m a t i o n c a n b e f o u nd a t : M a t c h m a ke r , h t t p ://w w w .c l o n t e c h .c o m ; H y b r i Z A P , h t t p ://w w w .g e n o m i c s .a g i l e n t .c o m ; P r o Q u e s t , h t t p s ://w w w .i n v i t r o g e n .c o m c p G B K T 7, t h e b a i t c l o n i n g v e c t o r (F i g . 2a ); p G B K T 7-p 53 a n d p G B K T 7-L a m , t h e c o n t r o l s p l a s m i d s (C l o n t e c h ) t h a t e x p r e s s t h e t u m o r s u p p r e s s o r p r o t e i n 53 a n d l a m i n p r o t e i n , r e s p e c t i v e l y d p G A D T 7, t h e p r e y c l o n i n g v e c t o r (F i g . 2b ); p G A D T 7-R e c , t h e p r e y l i b r a r y v e c t o r (F i g . 2b ); p G A D T 7-R e c T , t h e c o n t r o l p l a s m i d (C l o n t e c h ) w h i c h e x p r e s s e s t h e S V 40 v i r u s l a r g e T -a n t i g e n k n o w n t o i n t e r a c t w i t h p 53 b u t n o t w i t h L a m16911 Use of the Yeast Two-Hybrid System to Identify Targets of Fungal Effectors site (MCS) ﬂ anked by short stretches of sequence homologous tothe SMART (Switching Mechanism at 5 ¢ end of RNA Transcript) oligonuceotides that are designed for in vivo recombination ( 23) . Control plasmids pGBKT7-p53, pGBKT7-Lam, and pGADT7-RecTare also listed in Table1 . 1.A H109, the recipient yeast strain for expression or coexpres-sion of the fusion proteins: M ATa, trp1-901 , l eu2-3,112,ura3-52, his3-200, gal4 D , g al80 D , L YS2 :: GAL1 UAS - G AL1 TATA- H IS3, GAL2 UAS - G AL2 TATA - A DE2, URA3 :: MEL1 UAS- M EL1 TATA - l acZ, MEL1 ( s ee N ote 1 ). 2.Y 2HGold, the alternative recipient strain for expression or coexpression of the fusion proteins: M ATa, trp1-901, leu2-3, 112, ura3-52, his3-200, gal4 D , gal80 D , LYS2 : : GAL1 U AS –Gal1 T ATA –HIS3, GAL2 U AS –Gal2 T ATA –ADE2, URA3 : : MEL1 U AS –Mel1 T ATA -AUR1-C, MEL1 ( s ee N ote 2 ). 3.Y 187, the library strain and a mating partner with AH109 or Y2HGold: M AT a , l eu2-3,112, ura3-52, trp1-901, his3-200,ade2-101, gal4 D , g al80 D , m et−, URA3 :: GAL1 UAS - G AL1 TATA- l acZ , M EL1 ( s ee N ote 3 ).1.10 N NaOH solution. Dissolve 40 g sodium hydroxide pellets in 80 mL of ddH 2 O . Bring volume to 100 mL with ddH 2 O . Do not autoclave. 2.10× TE buffer: 0.1 M Tris–HCl, pH 7.5, 10 mM EDTA. Adjust pH to 7.5 and autoclave. 3.10× LiAc: 1 M lithium acetate. Adjust pH to 7.5 with dilute acetic acid and autoclave.2.1.2.Yeast Strains2.2.Buffersand SolutionsHAc-Myc*I lI II/Xma I I HI IMCS I I II/Xma I HI I IIMCS F ig. 2. M ain features of pGBKT7 ( a ), pGADT7, and pGADT7-Rec ( b ) (adapted from w ).pGADT7andpGADT7-Rec are the same except that the later has the SMART III and CDS III oligonucleotide sequences ( a sterisks )which are incorporated at the MCS for in vivo recombination ( 23).P A DH1 S . cerevisiae ADH1 promoter; T A DH1 S accharomyces cerevi-siae ADH1 terminator; P T 7 T7 RNA polymerase promoter; NLS SV40 nuclear localization signal; HA HA epitope tag; c -Mycc-Myc epitope tag; A mp r Ampicillin resistance gene; K anr kanamycin resistance gene; 5 ¢ A D and 3 ¢AD pGADT7-speciﬁc primers; 5 ¢ B D and 3 ¢BD pGBKT7-speciﬁc primers.170S. Lu4.50% PEG: Resuspend 25 g polyethylene glycol (MW = 3,350) in ~20 mL ddH 2 O . Bring volume up to 50 mL with ddH 2O and sterilize through a 0.45- m m syringe ﬁ lter ( s ee N ote 4 ).5.F ish sperm DNA (single-stranded DNA fragments): 10 mg/mL in 1× TE buffer, denatured by boiling for 10 min and immedi-ately placed on ice. Divide to aliquots in 1.5-mL microcentri-fuge tubes and store at −20°C. 6.P otassium phosphate solution (67 mM): Dissolve 0.91 g KH 2 P O 4 in ~80 mL of ddH 2O . Adjust pH to 7.5, bring volume up to 100 mL with ddH 2O , sterilize through a 0.22- m m syringe ﬁ lter, and store at 4°C for up to 1 year. 7. L yticase stock solution: 5 units/ m L lyticase in 1× TE buffer. Store at −20°C. 8. Z -buffer: D issolve 16.1 g N a 2 H PO 4 ·7H 2 O , 5.5 g N aH 2 P O 4 ·H 2O , 0.75 g KCl, 0.246 g MgSO 4 ·7H 2 O in ~800 mL of ddH 2O . Adjust pH to 7.0, autoclave and store at room temperature for up to 1 year. 9. X -gal and X- a -gal stock solutions: Dissolve X-gal (5-bromo-4-chloro-3-indolyl- b -D -galactopyranoside) or X- a -gal (5-bromo-4-chloro-3-indolyl- a - D-galactopyranoside) in N , N dimethyl-formamide (DMF) at a ﬁ nal concentration of 20 mg/mL. Divide to aliquots in 1.5-mL microcentrifuge tubes and store in the dark at −20°C. 10.A ureobasidin A (AbA) stock solution: 0.1 mg/mL, dissolved in 100% ethanol, stored at 4°C. 11. Z -buffer/X-gal solution: 100 mL of Z-buffer, 0.27 mL of b -mercaptoethanol, 1.67 mL of X-gal stock solution (20 mg/mL). Mix well just before use. 12. 50% glycerol (autoclaved). 13. 100% DMSO (Dimethyl sulfoxide). 14. 3 M sodium acetate (pH 4.8). 15. 95% ethanol (chilled at −20°C). 16. 0.9% NaCl solution.1.Y PD and YPDA media (for 1 L): Dissolve 10 g yeast-extract, 20 g bacto-peptone, 0.1 g adenine hemisulfate (for YPDAonly) in ~800 mL of ddH 2O . Adjust pH to 5.6–5.8 with 1 N NaOH. Add 20 g agar (for plates only) and bring volume upto 950 mL with ddH 2O . Autoclave at 121°C for 15 min. Add 50 mL of 40% glucose after cooling down to 55°C in a water bath and mix well before pouring the plates ( s ee N ote 5 ). 2.S ynthetic dropout (SD) medium (for 1 L): Dissolve 1.7 g yeast nitrogen base without amino acids and ammoniumsulfate 5.0 g ammonium sulfate, 20 gD -Glucose, 0.60–0.69 g 2.3.Yeast Media17111 Use of the Yeast Two-Hybrid System to Identify Targets of Fungal Effectorsready-to-use dropout (DO) supplement ( s ee N ote 6 ), or 100 mL of appropriate 10× DO stock solution (see below) in~800 mL ddH 2O . Adjust pH to 5.6–5.8 with 1 N NaOH. Add 20 g agar (for plates only) and bring volume to 1 L with ddH 2O . Autoclave at 121°C for 15 min. 3. 10× DO stock solution (for 1 L): Dissolve 200 mg L -Adeninehemisulfate salt, 200 mgL -Arginine HCl, 200 mg L -Histidine HCl monohydrate, 1,000 mgL -Leucine, 300 mg L -Lysine HCl, 200 mgL -Methionine, 500 mg L -Phenylalanine, 2,000 mgL -Threonine, 200 mg L -Tryptophan, 300 mg L -Tyrosine, 200 mg L -Uracil, and 1,500 mg L -Valine in 1 L ddH 2O (omit one or more amino acids as needed to make a speciﬁ c 10× DO solution for desired SD medium, e.g., add all except forL -Leucine and L -Tryptophan to make a 10× –Leu/–Trp DO solution). Autoclave at 121°C for 15 min and store at 4°C for up to 1 year.4. S D + X- a -gal plates: Prepare appropriate SD agar plates as described above, spread 50 m L of X- a -gal stock solution onto the surface of the solidiﬁ ed agar plates (100-mm) just before use. Alternatively, add 1 mL of X- a -gal stock solution to 1 L of SD agar medium after autoclave (cooled down to 55°C) before pouring the plates.5.S D-aureobasidin A (AbA) plates: Prepare appropriate SD agar plates as described above and add 1 mL of AbA stock solution to 1 L of SD agar medium after autoclave (cooled down to 55°C) before pouring the plates ( s ee N ote 7 ). O ptional: Kanamycin (50–100 m g /mL) can be added to all yeast media to avoid bacterial contamination.M ost strains of S . cerevisiae grow optimally at 30°C. On nonselec-tive YPD or YPDA agar plates, single cells become visible colonies usually 2–3 days after plating. On selective SD media, colonies may appear 2–5 days after plating depending on the stringency of the selection and the yeast strains ( s ee N ote 8 ). For liquid cultures starting with a single colony (2–3 mm in diameter) with shaking at 150–200 rpm, 24–36 h of incubation is sufﬁ cient to reach stationary growth stage.F or a short-term storage, yeast cells can be maintained on agar plates (tightly wrapped with Paraﬁ lm) and kept at 4°C for 1–2 months without losing viability ( s ee N ote 9 ). For long-term storage, yeast cells can be suspended in appropriate liquid media with 25% glycerol and kept in cryogenic vials at −80°C ( s ee N ote 10 ). To recover a3.Methods3.1.Growthand Maintenance of Yeast Strains3.1.1.General Growth Conditions3.1.2.Storageof Yeast Strains172S. Lustrain, streak a small amount of cells onto an YPD or YPDA plate and incubate at 30°C for 2–3 days. Before starting with transfor-mation or mating, verify growth phenotypes of the yeast strain by streaking the cells onto appropriate SD agar plates and allow growth at 30°C for 2–3 days.1. A mplify the gene of your interest ( G YI ) encoding the fungaleffector protein (not including the N-terminal signal peptide) by PCR using primers that contain 5 ¢ -end restriction sequencescompatible with the MCS on the bait vector pGBKT7 (Fig. 2a). Be sure that the G YI sequence to be cloned does not contain the restriction sites that have been incorporated at the 5 ¢ -ends of the PCR primers and will be translated in frame with the GAL4-BD domain. 2. P urify the ampliﬁ ed PCR product by phenol/chloroform extraction/ethanol precipitation or column puriﬁ cation. 3. D igest 1.0–3.0 m g of the puriﬁ ed PCR product using the restriction enzymes incorporated into the primer sequences under the conditions described by the enzyme suppliers (inac-tivate the restriction enzymes by heating or column puriﬁ cation after digestion).4.L igate the digested PCR product into pGBKT7 that has been linearized with compatible restriction enzymes using a T4 DNA ligase under required conditions.5. T ransform an E . coli strain with the ligated recombinant DNA(select for K anR ) and isolate the bait plasmid (pGBKT7-GYI) by column puriﬁ cation. 6. C onﬁ rm the sequence identity and the correct GAL4-BD-GYI fusion by DNA sequencing using pGBKT7-speciﬁ c primers 5 ¢ B D (5 ¢ -TCATCGGAAGAGAGTAGT-3 ¢ ) and 3 ¢ B D (5 ¢ -AGAGT-CACTTTAAAATTTGTAT-3 ¢ ) (Fig.2a ). 1.I noculate 5 mL of YPDA medium in a 50-mL centrifuge tube with a single colony (2–3 mm in diameter) of the yeast strain (AH109 or Y2HGold). Incubate overnight at 30°C with shaking (200–250 rpm). 2. I noculate 50 mL of YPDA in a 200-mL ﬂ ask with 0.5 mL of the overnight culture from s tep 1 . Incubate at 30°C with shaking(200–250 rpm) for 3–4 h until the OD 600 = 0.4–0.6. 3.S pin down the cells in a 50-mL tube using a GSA rotor (or equivalent) at 1,000 × g for 5 min at room temperature. Discardthe supernatants and resuspend the cell pellet in sterile ddH 2 O . 4.S pin down the cells and resuspend the cell pellet in 1 mL of TE/LiAc solution (freshly prepared by mixing 5 mL of 10× TEand 5 mL of 10× LiAc with 40 mL of ddH 2O ). Use competent cells immediately to obtain best transformation efﬁ ciency (see N ote 11 ).3.2.Generation of theBait Construct and Expression Strain3.2.1.Construction of the Bait Plasmid3.2.2.Preparationof Yeast Competent Cells11 Use of the Yeast Two-Hybrid System to Identify Targets of Fungal Effectors173B oth AH109 and Y2HGold can be used for expression of the3.2.3.Yeast Transformationbait fusion construct or coexpression of the bait and the preyconstructs. Include aureobasidin A (AbA) in SD/-LTHA and SD/-LTHA + X- a-gal plates if Y2HGold is used ( s ee N ote 7 ).1. C ombine 0.1 m g each of the bait and the prey plasmid DNA(pGADT7 or pGADT7-RecT, Table 1) in a 1.5-mL microcentri-fuge tube for each transformation experiment as listed below:(a) p GBKT7-GYI only(b) p GBKT7-GYI + pGADT7(c) p GBKT7-p53 + pGADT7-RecT (positive control)(d) p GBKT7-Lam + pGADT7-RecT (negative control)(e) N o DNA (blank control)2. A dd 0.1 mg of the denatured single-strand ﬁ sh sperm DNA.3. A dd 0.1 mL of competent cells to each tube and mix well.4. A dd 0.6 mL of PEG/LiAc solution (freshly prepared bymixing 1 mL of 10× TE and 1 mL of 10× LiAc with 8 mL of50% PEG) to each tube and vortex at high speed for at least10 s ( s ee N ote 12 ).5. I ncubate at 30°C for 30 min (invert tube every 10 min).6. A dd 20 m L of DMSO. Mix well by gentle inversion. Do notvortex.7. I ncubate the tube in a 42°C water bath for 15–20 min.8. S pin down cells at 13,000 rpm in an Eppendorf centrifuge atroom temperature for 10 s and discard the supernatant.9. R esuspend the cell pellet in 0.5 mL of sterile 1× TE buffer.10. S pread 100 m L of the transformed cells onto SD/–Trp, SD/–Leu–Trp (-LT), and SD/–Leu–Trp–His–Ade (-LTHA) agarplates separately. Incubate the plates (face down) at 30°C for2–3 days.11. C ompare results of the ﬁ ve transformations. The yeast trans-formants expressing the bait fusion construct alone (Expt. 1)should grow only on the SD/–Trp plate. The yeast transfor-mants coexpressing the bait construct and the pGADT7 vector(Expt. 2) should grow on the SD/-LT plate but not on theSD/-LTHA plate (Fig. 3a), like the negative control (Fig. 3c).Any growth on SD/-LTHA plate in Expt. 2 will suggest thatthe bait fusion protein has “activated” the reporter genes onits own (“autoactivation,” s ee N ote 13 ). Also, make sure thatthe colonies on the SD/-LT plate (Fig. 3a, left) in Expt. 2 aresimilar in size to those of the controls (Fig. 3b,c, left). If thecolonies are signiﬁ cantly smaller, the expressed bait protein islikely “toxic” to the yeast, thus not suitable for further analysis( s ee N ote 14 ).174S. LuF ig. 3. S D agar plates showing the results of transformation of AH109 with a bait fusionconstruct and control plasmids. Plates were inoculated with 100 m L of the transformedcells and incubated at 30°C for 3 days. Transformants coexpressing a bait and prey plasmids( a–c, corresponding to Expts. 2–4 in Subheading 3.2.3) all grew on SD/-LT plates ( l eft,arrow indicates a single colony), but only those expressing the positive control constructs( b) were able to grow on SD/-LTHA plate ( r ight ). All transformants in Expts. 1–4 grewequally well on SD/–Trp plates which select for T RP1only (not shown). The blank controldid not produce any colonies as shown in ( d).12. I f no apparent “autoactivation” or toxicity is associated with thebait construct, pick up a single colony from the SD/–Trp platein Expt. 1 and streak onto fresh SD/–Trp plate. Incubate at 30°Cfor 2–3 days. Use the yeast cells from the streaked colonies tomake a glycerol stock and store at −80°C ( s ee N ote 10 ).13. P erform a western blot analysis to verify the expression of the fusion protein following standard procedures ( s ee N ote 15 ). T his section describes procedures for the construction of a “non-normalized” cDNA library. Key materials used for ﬁ rst-strand cDNA synthesis and double-stranded cDNA ampliﬁ cation are components of the “Mate & Plate” Library System (Clontech). A “normalized” cDNA library may be required if the target genes are known to be expressed at a low level. The construction of such a library involves special procedures that are not within the scope of this chapter. Existing protocols ( 24 ) may be followed if applicable. 1. G row the host plant in a greenhouse or a growth chamber under the conditions appropriate for the fungal pathosystem to be tested. 2. I noculate the plant with an effector-producing fungal isolate or inﬁ ltrate the plant with fungal culture ﬁ ltrates that contain the secreted effector protein, or with the puriﬁ ed effector protein (if available). Incubate the inoculated plants for a desired period of time. 3. C ollect plant materials at several time points, e.g., 4, 12, 24, and 48 h after inoculation or inﬁ ltration (be sure to include the point where the induced symptoms become visible). Freeze the collected materials immediately in liquid nitrogen. 4. E xtract mRNA or total RNA using standard protocols or an appropriate commercial kit ( s ee N ote 16 ). Keep the isolated RNA on ice and proceed to ﬁ rst-strand cDNA synthesis immediately. 1. S et up the reaction in a 0.2-mL PCR tube as follows: R NA (Containing 0.025–1.0 m g mRNA or 0.1–2.0 m g totalRNA): 1–3 m LC DS III oligo(dT) primer: 1 m Ld dH 2 O : 0–2 m LT otal volume: 4 m L2. I ncubate at 72°C for 2 min, then immediately on ice for ³ 2 min.3. A dd the following to the reaction tube and then incubate at42°C for 10 min:5× First-Strand Buffer: 2.0 m L20 mM DTT: 1.0 m L10 mM dNTP Mix: 1.0 m LM MLV Reverse Transcriptase: 1.0 m L4. A dd 1.0 m L of SMART III oligonucleotide to the reactionmixture and incubate at 42°C for 1 h in a PCR thermalcycler( s ee N ote 17 ).3.3.Constructionof a cDNA Library3.3.1.Isolation of RNAfrom Plant Tissues3.3.2.First-Strand cDNASynthesis5. A dd 1.0 m L of RNase H and incubate at 37°C for 20 min. The synthesized ﬁ rst-strand cDNA can be used immediately for PCR ampliﬁ cation (below) or stored at −20°C for later use. 1. S et up two reactions in 0.2-mL PCR tubes, each containing as follows: F irst-strand cDNA (see Subheading 3.3.2 ): 2 m L 10× Advantage 2 PCR Buffer: 10 m L 50× Advantage 2 Polymerase Mix: 2 m L 50× dNTP Mix: 2 m L 5 ¢ PCR Primer: 2 m L 3 ¢ PCR Primer: 2 m L 10× GC-Melt Solution: 10 m L d dH 2 O : 70 m L T otal volume: 100 m L 2. R un PCR at the following conditions: 95°C, 30 s; 30 cycles of 95°C, 10 s, 68°C, 6 min; 68°C, 5 min. 3. R un 7 m L of the PCR product from each sample on a 1.0–1.5% agarose gel. A smear of range 0.3–6.0 kb should be visible (Fig. 4 , lanes 1 and 3). 1. P repare two CHROMA SPIN™ TE-400 Columns (Clontech) following the manufacturer’s instructions.2. C arefully load the double-stranded (ds) cDNA sample fromabove to the center of the gel bed’s ﬂ at surface in each columnprepared in step 1 and centrifuge at 700 × g for 5 min.3. C ombine the ﬂ ow-through samples (containing ds cDNA)into a 1.5-mL microcentrifuge tube.4. A dd 1/10 volume of 3 M sodium acetate (pH 4.8) and 2.5volume of 95% ethanol (prechilled at −20°C). Mix well andincubate at −20°C for 1 h to overnight.5. C entrifuge at 13,000 rpm for 20 min at room temperature.Remove the supernatant and allow the pellet in the tube to air-dry for 10 min.6. R esuspend the pellet in 25 m L of dd H 2 O .7. R un 2 m L of the puriﬁ ed ds cDNA on a 1.0–1.5% agarose gel.The intensity of the DNA smear (Fig. 4 , lanes 2 and 4) shouldbe similar to that of the unpuriﬁ ed ds DNA (lanes 1 and 3).The total amount of the puriﬁ ed ds cDNA should be 2.0–5.0 m g( s ee N ote 18 ).8. P roceed to in vivo recombination with pGADT7-Rec asdescribed below or store at −20°C for later use.3.3.3.Ampliﬁcation ofDouble-Stranded cDNA3.3.4.Puriﬁcation ofDouble-Stranded cDNA。

水污染英文 ppt课件

The pollutants include grit, asbestos石棉, phosphates and nitrates, mercury, lead, caustic soda and other sodium compounds, sulfur and sulfuric acid, oils, and petrochemicals.
Household Chemicals
Example: substances used for houses clean: Soap and synthetic detergents Wax Paints and paint removers Bleaches Disinfectants消毒剂 Polishes
批注本地保存成功开通会员云端永久保存去开通
Water pollution
Water is Essential for Life
It covers 71% of the earth's surface and makes up 65 % of our bodies.
Without the seemingly invaluable compound comprised of hydrogen and oxygen, life on Earth would be nonexistent
Refineries 精炼厂 release dyes, oils, acids 盐水, sulfur compounds, and other wastes.
Chemicals plants produce a variety of waste materials.
Oil spills
Oil spills from tankers at sea or leaks from underground storage tanks on land are very difficult to control

中药化学专业单词

8-methoxy psoralen 8-甲氧基补骨脂素electron atmosphere 电子云false positive 假阳性1,4-cineol 1，4-桉油醇10- octadecenoic acid 10-十八碳烯酸10-deacetylbaccatin 10-去乙酰基巴卡亭13-methylpalmatine 去氢延胡索素16-hydroxytriptolide 16羟基雷公藤内酯醇2,3-dibenzyl butyrolactone lignans 2,3-二苄基丁内酯木脂素2,4,6-trinitrophenol 2,4,6-三硝基苯酚2,6-dichloroquinone-4-chloroimide 2,6-二氯苯醌氯亚胺2-deoxyribose 2-脱氧核糖2-hydroxylsugar 2-羟基糖4-aminoantipyrine 4-氨基安替比林4-demethyliridoids 4-去甲基环烯醚萜4-hydroxy benzaldehyde 4-羟基苯甲醛4-hydroxymethylphenyl-b-D-glucopyranoside 4-羟甲基苯-b-D-葡萄吡喃糖苷5-HT 5-羟色胺6’-O-methylhonokiol 6’-O-甲基和厚朴酚6-epiharpagide 6-表哈帕苷7β-O-ethylmorroniside 7β-O-乙基莫诺苷9,12-octadecadienoic acid 9,12-十八二烯酸（亚油酸）9-hexadecenoic acid 9-十六碳烯酸9-octadecenoic acid 9-十八碳烯酸（油酸）Aabdominal distension 中满，腹胀abate jaundice 退黄（疸）abdominal fullness and distention 脘腹胀满abietane 松香烷abietic acid 松香酸absorbent 吸附剂acacetin 刺槐素Acacia catechu 儿茶acacias 洋槐Acanthaceae 爵床科acanthopanax 刺五加属acetal 乙缩醛acetamide 乙酰胺acetate 乙酸盐a cetate-malonate 乙酸-丙二酸acetic acid 乙酸acetic anhydride 乙酸酐acetolysis 乙酰解acetone 丙酮acetonitrile 乙腈acetyl 乙酰基acetyl acetone 乙酰丙酮acetylated 乙酰化的acetyl coenzyme A 乙酰辅酶A acetylation 乙酰化作用acetyl ginsenoside乙酰人参皂苷acetylic 乙酰化的aching of the loins and knees 腰膝酸痛Achyranthes bidentata牛膝acicular crystal 针状结晶acid 酸acid anhydride 酸酐acidic 酸的acidic hydrolysis 酸水解acidity 酸性，酸度acidulate 酸化Aconiti Radix 川乌aconitic acid 乌头酸aconitine 乌头碱acrid 辛Actions & Indications 功能主治activated charcoal 活性炭aculeatiside 颠茄皂苷acute 急性的acute lower respiratory tract infection 急性下呼吸道感染acute promyelocytic leukemia 急性髓性白血病acutif oliside 尖叶丝石皂苷acutissimin 野牡丹鞣质acutissimin A 一种复合鞣质acyclic 链状acyclic diterpenoids 无环二萜acyclic 无环的adaptogen 适应原additives 添加剂adenosine 腺苷adenylic acid 腺苷酸adhesive 黏合剂adipocyte differentiation 脂肪细胞分化adiposity 肥胖症adjacent 毗邻的adjusting immune system 调节免疫系统adjustment 调整adrenal 肾上腺的adsorbability 吸附力adsorbent 吸附剂adsorption 吸附aegle marmelos 印枸桔aerial 地上的aesculetin 七叶内酯、秦皮乙素aesculin 七叶苷、秦皮甲素ɑ-eudesmol 桉醇affinity chromatography 亲和色谱aflatoxin 黄曲霉素agar 琼脂agaropectin琼脂胶agarose琼脂糖a-glucosidase a-葡萄糖苷酶aglycone 苷元a-glycosidase a-苷酶Ailanthus altissima 臭椿ajoene 大蒜烯Akebia quinata 木通alanine dehydrase 丙氨酸脱氢酶albiflorin 白芍药苷alcohol sugar 醇糖alcoholic glycoside 醇苷alcoholic hydroxyl 醇羟基alcoholysis 醇解alcoholyze 醇解aldehyde 醛aldobionic acid 醛糖二糖酸aldohexose 己醛糖aldohexoside己醛糖苷aldose 醛糖aldoside 醛糖苷algae 藻类algae 藻类植物algin 褐藻酸alginic acid 褐藻酸alicyclic 脂环族的，脂环的Aliphatic hydrocarbons 脂肪烃aliphatic 脂肪族的aliquots 部分的Alisma orientalis泽泻alisol 泽泻萜醇alizarin 茜草素，茜草色素alkali 碱alkalify 碱化alkaline 碱的alkaloid 生物碱alkane 烷烃alkanin 紫草素alkanoids 生物碱alkyl 烷基（的），烃基（的）allergic asthma 过敏性哮喘allergies 过敏症allergy 过敏，变态反应allicin 大蒜辣素alliin 蒜氨酸alliinase 蒜氨酸酶allitridin 大蒜素Allium葱属allocryptopine 别隐品碱allomatrine 别苦参碱allose 阿洛糖allyl isosulfocyanate 异硫氰酸烯丙酯almond 杏仁almond oil 杏仁油aloe-emodin 芦荟大黄素aloin 芦荟苷alopecia areata 斑秃alpha streptococcus 甲型链球菌alphabetical 按字母表顺序的aluminum (Al) 铝aluminum trichloride 三氯化铝alzheimer dementia (AD) 老年性痴呆ambergris 龙涎香amenorrhea and dysmenorrhea 闭经痛经amethystoidin A 香茶菜甲素amide 酰胺amino acid 氨基酸amino group 氨基amino sugar 氨基糖aminobenzyl 氨基苯基aminophylline 氨茶碱ammonia氨Ammonium ceric nitrate 硝酸铈铵amorphous powders 无定形粉末amorphous solid 无定形固体amorphous 非晶型的amygdalase 苦杏仁苷酶amygdalin 苦杏仁苷amygdaloside 苦杏仁苷amyl alcohol 戊醇amylase 淀粉酶amylopectin 支链淀粉amylose 直链淀粉anabasine 八角枫碱anacardiacea 漆树科anaesthetic 麻醉anagyrine 安那吉碱，臭豆碱analgesia 镇痛analgesic 止痛剂，止痛的analgesic 止痛剂analog 类似物analogue 类似物a-naphthol a-萘酚Andrographia paniculata穿心莲andrographolide 穿心莲内酯anemia 血虚萎黄，贫血症Anemarrhena asphodeloides知母anemarrhena 知母anesthesia 麻醉anethole 茴香醚anethole 茴香脑Angelica sinensis当归angelic acid 当归酸angelicin 异补骨脂素，白芷素angiosperm 被子植物Angiosperms 被子植物angletype 角型angstroms 埃（长度单位）angular methyl 角甲基anhydride 脱水物anhydrous 无水的aniline 苯胺anion 阴离子anion-exchange 阴离子交换体anisaldehyde sulphuric acid 茴香醛-硫酸anisaldehyde 茴香醛，对甲氧基苯甲醛anisatin 莽草毒素anisic acid 茴香酸，对甲氧基苯甲酸anisodine 樟柳碱aniso-hybridization 不等性Anogeissus latifolia 宽叶榆绿木anomer 端基异构体anomeric carbon 端基碳anomeric carbon 端基异构碳ansu 山楂antagonist 拮抗剂antagonistic 拮抗的，对抗的antagonized 拮抗的anthelminthic 驱虫anthocyanidins 花青素anthocyanins 花色素anthracene 蒽anthracenol glycoside 蒽酚苷anthranol 蒽酚anthraquinone 蒽醌anthrone 蒽酮anti-aging 抗衰老antiallergic 抗过敏药antianaphylaxis 抗过敏antiangiogenic 抗血管生成anti-arrhythmia 抗心律失常antiarthritic 抗关节炎的anti-asthma 平喘作用anti-asthmatic 平喘的antibacterial 抗菌的antibacterium 抗菌antibiosis 抗菌(作用)antibiotic 抗菌素，抗生素anticancer 抗癌anticancer 抗癌的anticoagulant 抗凝剂anticoagulation 抗凝血作用anticonvulsant 抗惊厥的antidiabetic activity 抗糖尿病活性antidiabetics 抗糖尿病药antidiarrhreal 止泻的anti-fatigue 抗疲劳antifertility 抗生育的antiflammatory 抗炎的antifungal 抗真菌的antiherpesvirus 抗疱疹病毒antihyperglycemic 降高血糖的antihypertensive 降压antihypolipidemic 降血脂的anti-inflammatory 抗炎的(药) anti-inflammatory 抗炎药antimalaria 抗疟anti-microbial 抗菌的antimony chloride 氯化锑antimony trichloride 三氯化锑anti-mutagenic 抗突变anti-nociceptive 镇痛antioxidant 抗氧化antioxidant 抗氧化，抗氧剂antioxidative 抗氧化的antipyretic 退热剂anti-radiation 抗辐射antirheumatic 抗风湿剂antisepsis 抗菌，防腐，消毒antiseptic 消毒antiseptics 防腐剂anti-stress应激antithrombin 抗凝血酶antitumor 抗肿瘤antitussive 止咳药antiulcer 抗溃疡的antivenin 抗蛇毒素antiviral 抗病毒的Antivirus 抗病毒apatites 磷灰石apiaceae 伞形科apigenin 芹菜素apiose 芹菜糖Apocynaceae 夹竹桃科apoptosis 细胞凋亡apparatus 容器approximation 近似值apricot 杏，杏仁，杏树aqueous 水的aqueous solution 水溶液arabinogalactan 阿拉伯半乳糖arabinogalactan 阿拉伯半乳聚糖arabinose 阿拉伯糖arabinosyl阿拉伯糖基Araceae 天南星科arachic acid 花生酸，二十烷酸arachidic acid 花生酸arachidonic acid 花生四烯酸Arachis hypogaea 落花生Araliaceae 五加科Aralia chinensis楤木arbutin 熊果苷arctigenin 牛蒡子苷元Arctii Fructus 牛蒡子arctiin 牛蒡子苷Arctium lappa 牛蒡arctostaphylos uva-ursi 熊果areca seeds 槟榔arecatannin 槟榔鞣质arecolidine 槟榔碱arginine 精氨酸armeniacae semen amarum 苦杏仁Armillaria mellea 蜜环菌arnidiol 阿里二醇aroma 香气，芳香气aromatic ring 芳香环aromaticity 芳香性arrow-root木薯arsenic 砷arsenic trioxide 三氧化二砷arteannuic acid 青蒿酸arteether 蒿乙醚artelinic acid 青蒿中的倍半萜artemether 蒿甲醚artemisane 艾蒿烷型Artemisia annua黄花蒿（青蒿）Artemisia capillaries thunb 茵陈蒿artemisia ketone 蒿酮Artemisia scoparia waldst. et kit. 茵陈artemisiae scopariae herba （yinchen）茵陈artemisinin 青蒿素artesunate 青蒿琥酯artesunic acid 青蒿琥珀酸arthralgia 关节痛articulation 关节artifact 人工产物arundoin 芦竹素asarone 细辛醚ascaridole 驱蛔素Asclepiadaceae 萝藦科ascorbic acid 抗坏血酸，维生素Cash灰分asparagines 天冬酰胺aspartic acid 天冬氨酸asperuloside 车前草苷asphaltene 沥青质assay 试验Asteraceae 菊科Asteropus sarasinosum 海棉Aster tataricus 紫菀asthma 哮喘Astragali Radix黄芪astragalin 紫云英苷astragaloside黄芪苷 / 黄芪甲苷astragalus 黄芪属Astragalus gummifer 胶黄芪Astragalus membranaceus 膜荚黄芪Astragalus membranaceus(Fisch.) Bge.var.mongholicus蒙古黄芪astringency 收敛astragenol 黄芪醇astringent 收敛剂asymmetric 不对称的atherosclerosis 动脉粥样硬化atom 原子atractylone 苍术酮atropa belladonna 颠茄atropine 阿托品atropine 阿托品Aurones 橙酮authentic crude drug 标准药材auxochrome 助色团auxtochrous group 助色团availability 可利用性axial bond 直立键azelaic acid 壬二酸azulenoids 奥类化合物Bbaccatin 巴卡亭Bacillus aeruginosus 绿脓杆菌Bacillus coli 大肠杆菌Bacillus typhi 伤寒杆菌bacterial dysentery 细菌性痢疾bacteriological 细菌学的bacteriostat 抑菌剂bael 印度枳baicalein 黄芩素baicalin 黄芩苷bakuchalcone 补骨脂呋喃查耳酮bakuchiol 补骨脂酚baptifoline 野靛叶碱，膺靛叶碱barbaloin 芦荟苷barbary wolfberry fruit 枸杞子barium钡barium hydroxide 氢氧化钡basic solution 碱溶液bassorin 黄芪胶糖batches 成批,分批bavachalcone 补骨脂查耳酮bavachinin 补骨脂甲素甲醚bavachromanol 补骨脂色酚酮bavachromene 补骨脂色烯查尔酮be bound up 与…… 连在一起be indigenous to 土生土长bearberry leaves 熊果叶belladonna 颠茄Benefiting补益benzaldehyde 苯甲醛benzene 苯benzene hexachloride 六六六，六氯化苯，六氯环己烷benzoic 苯甲酸的benzoquinone 苯醌benzylpaeoniflorin 苯甲酰芍药苷berbamine 小檗胺berberastine 5-羟基小檗碱berberidaceae 小檗科berberine 小檗碱berberrubine 小檗红碱bererine 黄连素betain 甜菜碱betaine 甜菜碱Betulaceae 桦木科Betulin 白桦酯醇betulinaldehyde 白桦酯醛Betulinic acid 白桦酯酸betulinicacid 白蜡脂酸bicuculline 荷包牡丹碱bicyclic diterpenoids 双环二萜bicyclic sesquiterpenoids 双环倍半萜bicyclic 双环的bidirectional双向的Biflavonoids 双黄酮类Bignoniaceae 紫葳科bile acid 胆汁酸biliary 胆汁的bilobalide 白果内酯bilobalide 银杏内酯Bilobetin 白果素binding agent 粘合剂bioactive action 生物活性作用biogenetic 生源的biological source 基原biomass 生物体biosynthesis 生物合成biosynthetic 生物合成的biotransformation 生物转化bisabolane 没药烷bisdesmosides 双糖链苷bisdesmosidic saponins 双糖链皂苷bitter 苦味bitter apricot seed 苦杏仁bitter gourd 苦瓜bitter principle 苦味素bitter principles 苦味素bitterness 苦味black soyabean spermoderm 黑大豆皮bleaching powder 漂白粉blistering 发泡blood deficiency and sallow yellow 血虚萎黄blood ejection and spontaneous external bleeding 吐血衄血blood stasis and amenorrhea 瘀血经闭Blumea balsmifera艾纳香body fluid 体液boiling water 沸水boraginaceae 紫草科borax type 硼酸型boric acid 硼酸borneol 冰片，龙脑borneol 龙脑bornylmagnolol 龙胞基厚朴酚boron trifluoride 三氟化硼bottleneck瓶颈bracken fern 羊齿植物bradycardic 减慢心率的branched-chain 支链brassia alba白芥子brassia nigra黑芥子breast 乳腺bridging atom桥接原子bromelin 菠萝蛋白酶bromine 溴bromo 溴代bromophenol blue 溴酚蓝bronchial asthma 支气管哮喘bronchial smooth muscle 支气管平滑肌bronchitis 支气管炎bronchus 支气管bronic acid 硼酸brucine 马钱子碱bufadienolide 蟾蜍甾二烯内酯buffer medium 缓冲介质building block 组成…...的基本单位Bulbus Allii 大蒜Bupleuri Radix 柴胡Bupleurum chinense柴胡Bupleurum scorzonerifolium 狭叶柴胡Burseraceae 橄榄科butanol 丁醇butanone 丁酮butyl acetate 乙酸丁酯butyl alcohol 丁醇butylidene 丁烯基C cadinene 荜澄茄烯cadinane 杜松烷cadinol 杜松醇cadmium 镉caffeetannins 咖啡鞣质caffeic acid 咖啡酸caffeotannic acid 绿原酸caffeoyl 咖啡酰基calcium carbonate 碳酸钙calcium hypochlorite 次氯酸钙calibration 校准calm panting and suppress cough 止咳平喘caloglossa 鹧鸪菜calycosin 毛蕊异黄酮camelliatannin 山茶素，山茶鞣质Campanulaceae 桔梗科campesterol 菜油甾醇camphane 莰烷型camphene 莰烯camphor 樟脑camphoric acid 樟脑酸camptothecine 喜树碱candidate 候选者cannabinoid 大麻素canophyllal 海棠果醛canophyllic acid 海棠果酸cantharidin 斑蝥素cantoniensistriol 广东相思子三醇carabranecarane 蒈烷型carbocation 碳正离子carbohydrate 碳水化合物carbon 碳carbon tetrachloride 四氯化碳carbonium ion 正碳离子carbonyl 羰基carbonyl group 羰基carboxyl 羧基carboxyl methyl 羧甲基carboxylate 羧酸盐(酯)carboxylic acid 羧基？？羧酸？？？？carbuncle 痈carcinogenic 致癌的cardenolide 强心甾烯内酯cardiac contractility 心肌收缩性cardiac glycoside, cardioactive glycosides 强心苷cardiac output 心输出量cardio and cerebral vascular 心脑血管cardiodynia 心痛，胸痛cardio-protective 保护心脏Cardioprotector 心脏保护剂cardiovascular 心血管的cardiovascular and cerebrovascular diseases 心脑血管疾病carminative 祛风剂carica papaya 番木瓜carotene 胡萝卜素carotenoid 类胡萝卜素carrageenan 角叉菜胶Carthamin 红花苷Carthamone 醌式红花苷Carthamus tinctorius 红花carvone camphane 葛缕樟烷carvone camphor 香芹樟脑carvone 藏茴香酮Caryophyllaceae 石竹科caryophyllene 丁香油烃caryophyllene 石竹烯cassia obtusifolia 决明子cassia oil 肉桂油cassiamine 山扁豆双醌castalagin 栗木鞣花素casticin 紫花牡荆素casuarictin 木麻黄鞣亭casuarinin 木麻黄鞣宁catalpol 梓醇catalysis 催化作用catalyze 催化cataract 白内障Catechin 儿茶素catechol 儿茶酚catechu 儿茶catechuic acid 儿茶酸，儿茶素category 类别cation阳离子cation-exchange 阳离子交换体Celastraceae 卫矛科cellulase 纤维素酶cellulose 纤维素central excitation 中枢兴奋central nervous system中枢神经系统centrifuge 离心centrifuging-sedimentation 沉降法cephalin 脑磷脂cephalotaxin 三尖杉碱ceratonia siliqua 长角豆cerebral ischemia 脑缺血cervical cancer 子宫颈癌C-glycosides 碳苷C-glycosidic ellagitannins C-苷逆没食子鞣质chalcone 查耳酮chamigrene 花柏烯charcoal 炭chebuloyl (Che) 诃子酰基chemical ionization 化学电离chemical ionization (CI) 化学电离chemical shift 化学位移chemisorption 化学吸收作用chenodeoxycholic acid, chenodiol 鹅去氧胆酸chenopodium 土荆芥油chestnut 栗子Chinese galls 五倍子Chinese herbal medicine 中草药Chinese magnoliavine 五味子Chinese patent medicine 中成药Chinese sumac 盐肤木chiral carbon 手性碳chirality 手性chitin 几丁质cholesterol 胆固醇chloride 氯化物chloroform 氯仿, 三氯甲烷chlorogenic acid 绿原酸chlorophyll 叶绿素chlorophyll 叶绿素chloroquine 氯喹cholagogue 利胆的choleresis 利胆choleretic 利胆剂cholesterol 胆固醇, 胆甾醇chondroitin sulfate 硫酸软骨素chondrus cryspus 皱波角叉菜chorioepithelioma 绒癌chromatography 色谱chromic acid 铬酸chromogenic 显色的，产色的Chromone 色原酮chromophore 发色团，生色团chromophoric group 发色基团chronic infantile convulsions 小儿慢惊风chronic liver disease 慢性肝病chronic nephritis 慢性肾炎chronic toxicity 慢性毒性chronic tracheitis 慢性气管炎chrysophanol 大黄酚chuanliansu,toosendanin 川楝素cinchona 金鸡纳树皮cineole 桉叶油素cineole, eucalyptol 桉油精cinnabar 朱砂cinnabaris 辰砂cinnamaldehyde 桂皮醛cinnamic acid 桂皮酸Cinnamomum cassia肉桂cinnamon bark 肉桂皮cinnamon 肉桂cinnamyl acetate 乙酸肉桂酯cinnantannincinncassiol 肉桂萜醇cinnzeylanine 乙酰桂二萜醇cinnzeylanol 桂二萜醇circular dichroism 圆二色性cirrhosis 肝硬化cis顺式的Cistaceae 半日花科citral 柠檬醛Citri Reticulatae Pericarpium 陈皮citric acid 柠檬酸citronella oil 香茅油citronellol 香茅醇citrous 柑橘属植物的citrus aurantium枳壳Citrus aurantiun 酸橙citrus limonis 柠檬树Citrus peels 橙皮Citrus reticulata橘citrus 柑橘类植物classification 分类clear and benefit the head, eyes, and throat 清利头目，利咽clear fever from deficiency 清虚热clear heat and cool the blood 清热凉血clear heat and resolve fire toxicity 清热解毒clear-cut 清晰的cleavage 裂解clematis 铁线莲clerodane 克罗烷clinic 临床上cloud density 电子云密度clove 丁香coagulation 凝血coarse polysaccharides ch in eesis (CPSC) 五味子多糖coarse powder 粗粉coating 包衣，涂料cocaine 可卡因cocarcinogenic 辅致癌的codeine 可待因coefficient 系数Coelenterate 腔肠动物coenzyme 辅酶cognitive 认知的colchicine 秋水仙碱cold fever 感冒发热collision 碰撞colloidal state 胶体状态colon bacillus 大肠杆菌colorectal cancer 结肠直肠癌colubrine 克鲁勃林columbamine 非洲防己碱column chromatography 柱色谱法column 柱coma 昏迷combretaceae 使君子科common camptotheca root 喜树根common coltsfoot flower 款冬花complex crystal 络合物结晶complex enzyme 复合酶complex tannins 复合鞣质compositae 菊科comprehensive 综合的compromise 折中方法，综合各因素方法concentrate 浓缩concentration 浓度concentrated acid 浓酸condensation reaction 缩合反应condensed tannins 缩合鞣质condenser 冷凝器confer on 授予conﬁguration 构型conformation 构象conical 园锥形的conifer 松柏类Coniine 毒芹碱conjugate acid 共轭酸conjugate 共轭conjugated effect 共轭效应conjugated system 共轭系统consecutive reaction 连续反应constipation 便秘constipation due to deficiency of blood and body-fluid 肠燥便秘constrain the lung and stabilize panting 敛肺平喘contraceptive 避孕的，避孕药contractility 收缩性，收缩力conventional 传统的，习惯的convergence 收敛，会聚cool the blood 凉血cooling 清凉co-polymerization 异分子聚合copper 铜Coptidis Rhizoma 黄连coptisine 黄连碱coriamyrtin 马桑毒素corilagin 柯里拉京，鞣云实精Cornus officinalis 山茱萸coronary 冠状动脉coronary artery 冠状动脉coronary heart disease 冠状动脉心脏病correctant 矫味剂correlation spectroscopy 化学位移相关光谱cortex 皮质cortical皮层的，皮质的corybulbine 紫堇球碱，紫堇鳞茎碱corydaline 紫堇碱Corydalis Rhizoma 延胡索corydalmine 紫堇单酚碱corylidin 双羟异补骨脂定corynebacterium diphtheriae 白喉杆菌cosmetics 化妆品cough and asthma 咳嗽气喘cough and copious phlegm 咳嗽痰多cough-supressing and dispelling phlegm 止咳祛痰coumarin 香豆素coumarin glycoside 香豆素苷coupling constant 耦合常数course wind and dissipate heat 疏散风热covalent bond 共价键coexist 共存crab 蟹cranial nerve 脑神经crataegus pinnatifida山楂crimson tongue and polydipsia 舌绛烦渴criteria 标准critical point 临界点cross-conjugated 交叉共轭的cross-linking 交联croton 巴豆crotonic acid 巴豆油酸croton oil 巴豆油croton tiglium巴豆crotonoside 巴豆苷Cruciferae 十字花科cryptoxanthin 隐黄质crystal 结晶体crystallization 结晶化Cucurbitaceae 葫芦科cucurbitacin Ⅰa 雪胆甲素cucurbitacin Ⅱb 雪胆乙素cucurbitacins 葫芦素类cucurbitane 葫芦素烷cucurbitine 南瓜子氨酸cultivated species 栽培种Cupressaceae柏科Cupresuflavone 柏黄酮Curcuma kwangsiensis广西莪术Curcuma phaeocaulis蓬莪术Curcuma wenyujin温郁金curcumenolactone 莪术酮内酯curcumenone 莪术酮curcumin 姜黄素curcumol 莪术醇curdione 莪二酮cyamopsis tetragonolobus 瓜尔豆cyanin 花色素苷cyanogenic glycoside 氰苷cyclanoline 轮环藤酚碱cyclase 环化酶cyclic 环状的Cyclization 环合cyclization 环化作用cycloalliin 环蒜氨酸cycloartane 环木菠萝烷 / 环阿屯烷cycloastragenol 环黄芪醇cycloeudesmol 环桉醇cyclogeraniane 环香叶烷型cycloheptatriene cation 环庚三烯正离子cyclohexane 环己烷cyclohexane 环己烷cyclohexene环己烯cyclohexylamines 环己胺cyclopentane 环戊烷cyclopentanoperhydrophenanthrene 环戊烷骈多氢菲cyclopropane 环丙烷cylindrical 园柱形的cylindrin 白茅素cymarose 加拿大麻糖Cyperaceae 莎草科cysteine sulfoxides 半胱氨酸亚砜cystine 胱氨酸cytidine 胞苷cytisus 金雀花cytotoxicity 细胞毒性Ddaidzein 大豆苷元daidzin 大豆苷，黄豆苷（异黄酮苷）dammarane 达玛烷damp-heat jaundice 湿热黄疸dampnessdaphane 瑞香烷Daphne genkwa芫花daphnetoxin 瑞香毒素daphnin 瑞香苷dark plum fruit 乌梅Daturae Flos 洋金花daturameteloside 洋金花苷dauco sterine 胡萝卜苷daucosterol 胡萝卜甾醇，胡萝卜苷daurian rhododendron oil 满山红油dauricine 蝙蝠葛碱decanoylacetaldehyde 癸酰乙醛decanta 泻下decarboxylate 脱羧decoction 煎煮decompose 分解decomposition 分解，变质decompression 降压decyl 癸基defatted 脱脂degradation 降解dehydration 脱水dehydrocholic acid 脱氢胆酸dehydrocorybulbine 去氢紫堇鳞茎碱dehydrocorydaline 去氢紫堇碱dehydrogenate 脱水dehydrogenation 脱氢作用dehydroglaucine 去氢海罂粟碱dehydrohexahydroxydiphenoyl (DHHDP) 脱氢六羟基联苯二甲酰基dehydronantenine 去氢南天宁碱20(21)-dehydroxylation-ginsenoside 20(21)-脱羟基-人参皂苷deionise 去离子dementia 痴呆demethylation ephedrine 去甲基麻黄碱demulcent 镇痛剂dencichine 三七素dendrobine 石槲碱dense 粘稠的，稠密的deodorant 除臭剂deohydroandrographolide 脱水穿心莲内酯deoxy 脱氧的deoxyandrographolide 去甲穿心莲内酯deoxycholic acid 去氧胆酸deoxyschisandrin 五味子甲素deoxysugar 去氧糖deprotonation 去质子化作用depside 缩酚酸derivative 衍生物，派生物dermatitis 皮炎dermatosis 皮肤病dermerethistica 皮肤刺激药Derris elliptica 毛鱼藤deshielding去屏蔽desinsection 杀虫determination 决定，鉴定detoxify 解毒detrimental 有害的，不利的detumescence 消肿deuterium 氘，重氢development system 展开系统dexiotropic 右旋的dextrorotation 右旋D-galacto-D-manoglycon D-半乳糖-D-甘露糖基D-galactouronic acid D-半乳糖醛酸diabetes 糖尿病diabetes caused by internal heat 内热消渴diacolation 渗滤dialcol 二醇dialdehyde 二醛dialysis 透析diameter 直径diarrhea 腹泻diastereoisomer 非对映异构体diastolize 舒张diazomethane 重氮甲烷diazonium 重氮化合物dibenzyl-etrahydroisoquinolin 双苯甲基四氢异喹啉类dichlorodiphenyltrichloroethane二氯二苯三氯乙烷，滴滴涕dichloromethane 二氯甲烷dicotyledon 双子叶植物diels-alder (D-A反应）双烯加成反应diene二烯（烃）dietary fiber 膳食纤维diethyl ether 乙醚diethylaminoethyl 二乙氨基乙基diffraction 衍射digestive system 消化道系统diginatigenin 双羟基洋地黄毒苷元digitalose 洋地黄糖digitoxigenin 洋地黄毒苷元digitoxin 洋地黄毒苷digitoxose 洋地黄毒糖digoxigenin 异羟基洋地黄毒苷元digoxin 地高辛dihydrochalcones 二氢查耳酮类dihydroflavonol 二氢黄酮醇dihydromorin 二氢桑色素dihydroquercetin 二氢槲皮素dihydrosanguinarine 二氢血根碱dilactone 双内酯dilatation 扩张dilate 扩大，膨胀dilute acid 稀酸dilute alcohol 稀醇dilute alkali solution 稀碱液dilute base 稀碱Dimeric 二聚的dimers 二聚体dimethoxy 二甲氧基dimethyl formamide 二甲基甲酰胺dimethyl sulphate 硫酸二甲酯dimethyl sulphoxide 二甲基亚砜dimethylallyl pyrophosphate 二甲基烯丙酯p-dimethylamino benzaldehyde 对二甲氨基苯甲醛dimethylaniline 二甲基苯胺dimethylformamide 二甲基甲酰胺dimethylpolysiloxane 二甲硅油dimethyltetrandrine 二甲基粉防己碱Dioscoreaceae 薯蓣科Dioscorea 薯蓣属Dioscorea nipponica 穿龙薯蓣Dioscoreae Nipponicae Rhizoma 穿山龙dioscin 薯蓣皂苷diosgenin 薯蓣皂苷元diosmin 香叶木苷Diospyros柿属Diospyros kaki柿子dipiperitylmagnolol 双辣薄荷基厚朴酚diploptene 里白烯dipole-dipole interactions 偶极偶极相互作用disaccharide 二糖discrete 分离的，不相关联的disintegrate 裂解disintegrating agent 崩解剂d-isomer 右旋异构体dispel wind and relieve pain 祛风止痛disperse accumulations and relieve pain 消积止痛disperse swelling 消肿disperse 分散，散开dissolved 溶解dissolution 溶解distending pain胀痛distention and oppression in the chest and rib-side 胸胁胀闷Distortion enhancement by polarization transfer (DEPT) 极化转移增强distribution constant 分配系数diterpenoid 二萜diuresis 利尿，多尿diuretic 利尿剂diuretics 利尿剂，利尿药divinylbenzene 二乙烯苯dizziness and palptation 眩晕心悸dizziness and tinnitus 眩晕耳鸣doesahexaenoic acid 二十二碳六烯酸dopamine 多巴胺dotriacontane 三十二烷double lignans 双木脂素Double Quantum Filtered 1H-1H COSY (DQF-COSY) 双量子H-H 相关谱down-field 低磁场drain summer heat 解暑热drift 迁移drinking and urine 消渴dry dampness and transform phlegm 燥湿化痰Dryobalanops camphora龙脑香树Dryopteris crassirhizoma 贯众dual-modulation 双向调节ducitol 卫矛醇duct 导管duodenal tumours 十二指肠癌dysentery 痢疾dysmenorrheal 痛经dyspepsia 消化不良dyspnea and coughing 胸闷咳嗽Eechinacea purpurea紫花松果菊echinacea 紫锥菊eclampsia 子痫，惊厥eco-friendly 环境友好型的ecuelle 压榨法eczema 湿疹eduasarone 欧细辛醚eicosanoic acid 花生酸eicosapentaenoic acid 二十碳五烯酸electric field 电场electrodialysis 电渗析electrolysis 电泳electron donating 供电子electron impact 电子碰撞电离electron impact (EI) 电子轰击electronegativity 电负性electron-withdrawing 吸电子electrophilic 亲电子的，吸电子的electrophilic group 亲电基团electrophoresis 电泳electrospray ionization 电喷射电离electrospray ionization (ESI) 电喷雾电离elemane 榄香烷elevating 提升,提高eliminate dampness 排除湿气eliminating phlegm 排除痰液eliminated 消除的elimination 消除ellagitannins 逆没食子鞣质，鞣花酸鞣质eluate 洗脱液Embelia oblongifolia多脉酸藤子Embelia ribes 白花酸藤果emetine 吐根碱emmenagogue 调经药emodin 大黄素empirical 经验的emulgent 乳化剂emulsifier 乳化剂emulsin 苦杏仁酶emulsion乳汁/ 乳状液emulsion 苦杏仁苷酶enantiomer 对映体endocrine system 内分泌系统endoperoxide 内过氧化物endothelial 内皮的enfleurage 吸香法enol 烯醇enolic glycoside 烯醇苷enuresis 遗尿enzymatic 酶的enzymatic hydrolysis 酶水解enzymatic specificity 酶特异性enzyme 酶enzymolysis 酶解Ephedraceae 麻黄科Ephedrae Herba 麻黄ephedrine hydrochloride 盐酸麻黄碱ephedroxane 麻黄噁唑酮epiberberine 表小檗碱epicatechin 表儿茶素Epicatechin 表儿茶素epichlorohydrin 表氯醇epigastric腹上部的epimer 差向异构体epimeric structure 端基异构体epoxide ring 环氧环epoxide 环氧化物epoxy 环氧的epoxy 环氧基equatorial bond 平伏键equilibrium 平衡Eremostachys沙穗属ergosterol 麦角甾醇ergot 麦角，麦角菌Ericaceae 杜鹃花科Ericaceae 杜鹃花科eriocitrin 圣草次苷Erodii Herba 老鹳草erysipelas 丹毒erythrina species刺桐属esculentic acid 商陆酸esculentoside 商陆皂苷esculetin 七叶内酯，秦皮乙素esculin 七叶苷，秦皮甲素essential amino acids 必需氨基酸essential oil 精油ester 酯ester glycoside 酯苷ester linkage 酯键esterification 酯化ester saponins 酯皂苷estrogen 雌性激素Estrogen receptor 雌激素受体Estrogenic 雌激素的ethanol 乙醇ethereal oil 香精油，醚油（醚和杂醇油的混合物）etherification 醚化作用ethyl acetate 乙酸乙酯ethylene glycol 乙二醇eucalyptus oil 桉油eudesmane 桉烷eudesmol 桉醇，桉叶油醇eugenol 丁香酚eugenol type basil oil 丁香罗勒油euparotin 泽兰苦内酯Eupatorium rotundifolium圆叶泽兰Euphane 大戟烷Euphorbia大戟属Euphorbia fischeriana狼毒大戟Euphorbia kansui甘遂Euphorbia lathyris千金子Euphorbiaceae 大戟科eutectic point 共熔点evodiamine 吴茱萸碱excitatory有刺激性的，兴奋的excreting dampness 渗湿exhaustive 全面的，彻底的exocyclic 环外的expectorant 祛痰剂expelling 驱逐explosive 易爆炸的extinction coefficient 消光系数extract 提取物extraction 提取extremities sores 四肢疼痛exudate 渗出物F Fabaceae 豆科Faboideae 豆科蝶形花亚科Fagaceous plant 山毛榉科植物false negative 假阴性Fam. (familiar) 科fangchinoline 汉防己乙素，防己诺林碱faradiol 款冬二醇farnesane 金合欢烷farnesol 金合欢醇farnesyl pyrophosphate, FPP 焦磷酸金合欢酯fast atom bombardment (FAB) 快原子轰击fatty acid 脂肪酸fenchane 葑烷型fermentation 发酵fernane羊齿烷fernenol 羊齿烯醇Ferns 蕨类ferric chloride 氯化铁ferric trichloride 三氯化铁fertility 生育力，生育率fertilization 受精ferulic acid 阿魏酸feruoyl 阿魏酰基fever due to deficiency of Yin 阴虚发热fever from summer heat evil 暑邪发热fibrous root 须根Ficine 黄酮榕碱field desorption (FD) 场解吸法field desorption ionization 场解析电离field ionization 场致电离figwort root 玄参filariasis 丝虫病filter 过滤filtrate 滤液fire-purging drugs 清火药five watches 五更flammability 易燃，可燃性flavan-3,4-diol 黄烷-3,4-二醇Flavan-3,4-diols 3,4-二羟基黄烷醇flavan-3-ol 黄烷-3-醇Flavanol 黄烷醇Flavanones 黄烷酮类Flavanonols 二氢黄酮醇类Flavans 黄烷flavanyl 黄烷基Flavones 黄酮flavono-ellagitannins 黄酮类-鞣花酸鞣质flavonoid 黄酮flavonoid glycoside 黄酮苷Flavonoids 黄酮类Flavonols黄酮醇类Flavylium 花色基元flow chart 流程图fluid deficiency and constipation 津伤便秘fluorescence 荧光性foam 泡沫Foeniculum vulgare小茴香follicle hyperkeratosis 角化过度food accumulation and distending pain 食积胀痛formamide 甲酰胺formic acid 甲酸formidable 可怕的，令人畏惧的formononetin 刺芒柄花素Forsythia 连翘，连翘属植物Forsythia suspensa连翘forsythiaside 连翘酯苷forsythol 连翘酚fractionation 分馏法，分别fragment碎片fragmentation 碎片,破碎fragmentation mechanism 碎片裂解机制fragrance 芳香气味frankincense 乳香fraxetin 秦皮素fraxin 秦皮苷Fraxini Cortex 秦皮Fraxinus 白蜡树属fraxinus ornus花白腊木fraxinus rhynchophylla 苦枥白蜡树freckle 雀斑，斑点free fatty acid 游离脂肪酸free hydroxyl 游离羟基free radical 自由基free the network vessels and relieve pain 通络止痛friedelane木栓烷friedelin 木栓酮fritillaria tunbergii var. chekiangensis 东贝母frozen 凝固；冷冻fructan 果聚糖fructo-furanose unit 果糖呋喃糖单位fructose 果糖fructoside 果糖苷frufuranose 呋喃果糖fucose 夫糖, 岩藻糖fucoside 夫糖苷fucosterol 岩藻甾醇functional group 官能团fungus（fungi）真菌funnel 漏斗furan ring 呋喃环furanose 呋喃糖furanoside 呋喃糖苷furanspongin-3 呋喃海绵素-3furocoumarin 呋喃香豆素furostanol 呋甾烷醇fused 稠合fused-ring 稠环fused 乌本苷/G-毒毛花苷Ggalactomannan 半乳甘露聚糖galactose 半乳糖galactoside 半乳糖苷galactosyl 半乳糖基galacturonic acid 半乳糖醛酸galangin 三羟基黄酮gall aphid 五倍子蚜Galla Chinensis 五倍子gallate 没食子酸盐gallbladder 胆囊gallotannins 没食子鞣质galloyl esters 没食子酸酯galloylglucoses(GGs) 没食子酰基葡萄糖Calycosin-7-glucoside毛蕊异黄酮-7-葡糖苷ganoderma lucidum灵芝gas chromatography 气相色谱gas liquid chromatography 气液色谱gastric mucosa 胃粘膜gastrodia elata 天麻gastrodin 天麻苷gastroenteritis and dysentery 肠胃炎及痢疾gastroenteritis 胃肠炎gelidiaceae 石花菜科gelidium amansii 石花菜general debility with deficiency of vital essence 虚劳精亏genin 苷元genipin-1-O-gentiobioside 京尼平-1-O-龙胆双糖苷geniposide 京尼平苷Gentianaceae 龙胆科gentianine 龙胆碱Gentianose 龙胆三糖gentiobiose 龙胆双糖gentiopicroside,gentiopicrin 龙胆苦苷genus 属geometric 几何的geraniaceae 牻牛儿苗科geranial 香叶醛Geranii Herba 老鹳草geraniin 老鹳草素geraniol 香叶醇Geranium macrorrhizum大根老鹳草geranyl acetate 乙酸香叶酯geranyl pyrophosphate, GPP 焦磷酸香叶酯geranylfarnesyl pyrophosphate, GFPP 焦磷酸香叶基金合欢酯geranylgeranyl pyrophosphate, GGPP 焦磷酸香叶基香叶酯germacrane 吉马烷germacrone 吉马酮Gesneriaceae 苦苣苔科gigarginaceae杉藻科gigartina sps 杉藻gingkgolide 银杏内酯Ginkgetin 银杏素Ginkgo biloba 银杏ginkgo biloba 银杏科ginkgo folium (yinxingye) 银杏叶Ginkgoaceae 银杏科ginkgolide 银杏内酯ginkgolides 银杏内酯ginseng 人参ginsenoside 人参皂苷Ginseng Radix et Rhizoma 人参gitaloxigenin 吉他洛苷元gitoxigenin 羟基洋地黄毒苷元gitoxin 羟基洋地黄毒苷glacial 冰的，冰样的globular protein 球状蛋白。

天然药物化学专业英语词汇总结

专业英语词汇总结Section 1生药部分中药研究现状及中药现代化一、加强中国药用植物基础研究及其与中药现代化的联系/Strengthening basic researches on Chinese Medicinal Plants and its relations to realizing the modernization of CMM记载be recorded来源derived from中医药Traditional Chinese Medicine,short for TCM卫生事业health care,health undertakings中草药Chinese traditional medicinal herbs疗效reliable therapeutical effectstherapeutic[,θer?'pju:t?k]adj.治疗(学)的;疗法的；对身心健康有益的副作用side-effectsl中医药的健康理念和临床医疗模式体现了现代医学的发展趋势。

The health concept and clinical practice reflect the trend of modern science新的科学技术潮流（the new tide of science and technology）二、中药资源及其研究成果/Chinese Medicinal Plant resources and achievement of its scientific research中药资源(medicinal plant resources)普查(surveys)专项研究(special projects)药用植物资源(the Chinese medicinal resources)科学鉴定(scientific identification)化学成分(chemical constituents)药理实验(pharmacological experiments临床适应症(clinical applications)研究(projects)新著作(new works)各论(monographs)手册(manuals)《中国药典》The pharmacopoeia of the people’s Republic of China药典Pharmacopoeia药用植物学Pharmaceutical Botany本草学Herbology中药学The Chinese Materia Medica药用植物分类学Pharmaceutical Plant Taxonomy植物化学Phytochemistry植物化学分类学Plant Chemotaxonomy药用植物志Flora of Medicinal Plant中药药剂学traditional Chinese Pharmaceutics中药炮制学Science of processing Chinese Crude Drugs中药鉴定学Identification of Traditional Chinese Medicine中药药理学Pharmacology of Traditional Chinese Medicines青蒿素artemisin奎宁quinine、氯奎宁chloroquine衍生物derivatives氯奎宁耐受性疟疾chloroquine resistant malaria急性疟疾pernicious malaria脑部疟疾cerebral malaria显著疗效marked effect chloroquine resistant malaria/抗氯喹啉疟疾Pernicious（有害的）malaria/急性疟疾cerebral malaria/脑疟疾derivatives/衍生物quinine/喹啉含有氮原子的化合物，在英文命名中多以-ine结尾Mono-/一Di-/二Tri-/三Tetra-/四Petan-/五Hexa-/六Hepta-/七Octa-/八Nona-/九Deca-/十三尖杉酯碱harringtonine、高三尖杉酯碱homoharringtonine白血病leukemia和恶性淋巴瘤malignant lymphoma银杏黄酮ginkgetin丹参酮tanshinon IIA治疗冠心病coronary heart diseasesNew drug developments/新药开发Health products/保健品质量控制Quality control修订revise常用中药common-used Chinese materia medica国家标准the national standards三、中药所面临的挑战/Chinese Medicinal Herbs Facing a Challenge中成药及其制剂traditional Chinese patent medicines and preparations基础研究basic researches生产production、流通marketing研究researchIdentification of species/品种鉴定鉴定和鉴别identifying and clarifying变种varieties伪品false matters。

双语教学中的生物化学词汇

双语生物化学词汇Glossary of Biochemistry BilinguallyAAbsolute configuration（绝对构型）The configuration of four different substituent groups around an asymmetric carbon atom, in relation to u- and i.-glyceraldehyde. Absorption（吸收）: transport of the products of digestion from the intestinal tract into the blood.Acceptor control（受体控制）:The regulation of the rate of respiration by the availability of ADP as phosphate group acceptor.Accessory pigments（辅助色素）:Visible light-absorbing pigments (carotenoids, xanthophyll, and phycobilins藻胆素) in plants and photosynthetic bacteria that complement chlorophylls in trapping energy from sunlight.Acidosis（酸中毒）: A metabolic condition in which the capacity of the body to buffer is diminished; usually accompanied by decreased blood pH.Actin（肌动蛋白）: A protein making up the thin filaments（细丝）of muscle; also an important component of the cytoskeleton of many eukaryotic cells.Activation energy(ΔG*)（活化能）: The amount of energy (in joules) required to convert all the molecules in 1 mole of a reacting substance from the ground state to the transition state.Activator:（活化物、激活剂）(1) A DNA-binding protein that positively regulates the expression of one or more genes; that is, transcription rates increase when an activator is bound to the DNA. (2) A positive modulator of an allosteric enzyme.Active site:（活性部位）The region of an enzyme surface that binds the substrate molecule and catalytically transforms it; also known as the catalytic site.Active transport:（主动运输）Energy-requiring transport of a solute across a membrane in the direction of increasing concentration.Activity:（活度）The true thermodynamic activity or potential of a substance, as distinct from its molar concentration.Activity coefficient:（活度系数）The factor by which the numerical value of the concentration of a solute must be multiplied to give its true thermodynamic activity. Adenosine 3',5'-cyclic monophosphate: See cyclic AMP.Adenosine diphosphate: See ADP.Adenosine triphosphate: See ATP.Adipocyte:（脂肪细胞）An animal cell specialized for the storage of fats (triacylglycerols).Adipose tissue:（脂肪组织）Connective tissue specialized for the storage of large amounts of triacylglycerols.ADP (adenosine diphosphate):A ribonucleoside diphosphate serving as phosphate group acceptor in the cell energy cycle.Aerobe:（需氧生物）An organism that lives in air and uses oxygen as the terminal electron acceptor in respiration.Aerobic: Requiring or occurring in the presence of oxygen.Alcohol fermentation:（乙醇发酵）The anaerobic conversion of glucose to ethanol via glycolysis. See also fermentation.Aldose:（醛糖）A simple sugar in which the carbonyl carbon atom is an aldehyde; that is, the carbonyl carbon is at one end of the carbon chain.Alkalosis:（碱中毒）A metabolic condition in which the capacity of the body to buffer is diminished; usually accompanied by an increase in blood pH.Allosteric enzyme:（变/别构效应） A regulatory enzyme, with catalytic activity modulated by the noncovalent binding of a specific metabolite at a site other than the active site.Allosteric protein: （变/别构蛋白）A protein (generally with multiple subunits) with multiple ligand-binding sites, such that ligand binding at one site affects ligand binding at another.Allosteric site: （变/别构部位）The specific site on the surface of an allosteric enzyme molecule to which the modulator or effector molecule is bound.α helix:（α-螺旋）A helical conformation of a polypeptide chain, usually right-handed, with maximal intrachain hydrogen bonding; one of the most common secondary structures in proteins.Ames test:A simple bacterial test for carcinogens, based on the assumption that carcinogens are mutagens.Amino acid activation:（氨基酸活化）ATP-dependent enzymatic esterification of the carboxyl group of an amino acid to the 3'-hydroxyl group of its corresponding tRNA. Amino acids:（氨基酸）an Amino-substituted carboxylic acids, the building blocks of proteins.Amino-terminal residue:（氨基末端残基）The only amino acid residue in a polypeptide chain with a free a-amino group; defines the amino terminus of the polypeptide.Aminoacyl-tRNA:（氨酰tRNA）An aminoacyl ester of a tRNA.Aminoacyl-tRNA synthetases:（氨酰tRNA合成酶）Enzymes that catalyze synthesis of an aminoacyl-tRNA at the expense of ATP energy.Aminotransferases:（氨基转移酶）Enzymes that catalyze the transfer of amino groups fromα-amino to α-keto acids; also called transaminases.Ammonotelic:（排氨的）Excreting excess nitrogen in the form of ammonia. Amphibolic pathway:（双向代谢途径）A metabolic pathway used in both catabolism and anabolism.Amphipathic:（双亲的）Containing both polar and nonpolar domains. Ampholyte:（两性电解质）A substance that can act as either a base or an acid. Amphoteric:（两性的）Capable of donating and accepting protons, thus able to serve as an acid or a base.Anabolisim:（合成代谢）The phase of intermediary metabolism concerned with the energy-requiring biosynthesis of cell components from smaller precursors. Anaerobe:（厌氧生物）An organism that lives without oxygen. Obligate anaerobes （专性厌氧生物）die when exposed to oxygen.Anaerobic:（厌氧的）Occurring in the absence of air or oxygen.Anaplerotic reaction:（回补反应）An enzyme-catalyzed reaction that can replenish the supply of intermediates in the citric acid cycle.A ngstrom (Ǻ):（唉）A unit of length (10-8cm) used to indicate molecular dimensions. Anhydride:（酸酐）The product, of the condensation of two carboxyl or phosphate groups in which the elements of water are eliminated to form a compound with the general structure R—X—0—X—R, where X is either carbon or phosphorus.Anion-exchange resin:（阴离子交换树脂）A polymeric resin with fixed cationic groups; used in the chromatographic separation of anions.Anomers:（异头物、端基异构体）Two stereoisomers of a given sugar that differ only in the configuration about the carbonyl (anomeric) carbon atom.Antibiotic:（抗生素）One of many different organic compounds that are formed and secreted by various species of microorganisms and plants, are toxic to other species, and presumably have a defensive function.Antibody:（抗体）A defense protein synthesized by the immune system of vertebrates. See also immunoglobulin.Anticodon:（反密码子） A specific sequence of three nucleotides in a tRNA, complementary to a codon for an amino acid in an mRNA.Antigen:（抗原）A molecule capable of eliciting the synthesis of a specific antibody in vertebrates.Antiparallel:（反平行）Describing two linear polymers that are opposite in polarity or orientation.Antiport:（反向转运）Cotransport of two solutes across a membrane in opposite directions.Apoenzyme:（酶蛋白）The protein portion of an enzyme, exclusive of any organic or inorganic cofactors or prosthetic groups that might be required for catalytic activity. Apolipoprotein:（脱辅基脂蛋白）The protein component of a lipoprotein. Apoprotein: （脱辅基蛋白）The protein portion of a protein, exclusive of any organic or inorganic cofactors or prosthetic groups that might be required for activity. Apoptosis:（细胞凋亡）(app'-a-toe'-sis) Programmed cell death, in which a cell bringsabout its own death and lysis, signaled from outside or programmed in its genes, by systematically degrading its own macromolecules.Arrestin:（抑制蛋白） A family of proteins that bind to the phosphorylated carboxyl-terminal region of serpentine receptors, preventing their interactions with G proteins and thereby terminating the signal through those receptors.Asymmetric carbon atom:（不对称碳原子）A carbon atom that is covalently bonded to four different groups and thus may exist in two different tetrahedral configurations. ATP (adenosine triphosphate): A ribonucleoside 5'-triphosphate functioning as a phosphate group donor in the cell energy cycle; carries chemical energy between metabolic pathways by serving as a shared intermediate coupling endergonic and exergonic reactions.ATP synthase:（ATP合酶）An enzyme complex that forms ATP from ADP and phosphate during oxidative phosphorylation in the inner mitochondrial membrane or the bacterial plasma membrane, and during photophosphorylation in chloroplasts. ATPase:（ATP酶）An enzyme that hydrolyzes ATP to yield ADP and phosphate; usually coupled to some process requiring energy.Attenuator:（弱化子）An RNA sequence involved in regulating the expression of certain genes; functions as a transcription terminator.Autotroph:（自养生物）An organism that can synthesize its own complex molecules from very simple carbon and nitrogen sources, such as carbon dioxide and ammonia. Auxin:（植物生长素）A plant growth hormone.Auxotrophic mutant (auxotroph):（营养缺陷突变体）A mutant organism defective in the synthesis of a given biomolecule, which must therefore be supplied for the organism's growth.Avogadro's number: The number of molecules in a gram molecular weight (a mole) of any compound (6.02 × 1023).BBack-mutation:（回复突变）A mutation that causes a mutant gene to regain its wild-type base sequence.Bacteriophage (phage):（噬菌体）A virus capable of replicating in a bacterial cell. Basal metabolic rate:（基础代谢率）The rate of oxygen consumption by an animal's body at complete rest, long after a meal.Base pair:（碱基对）Two nucleotides in nucleic acid chains that are paired by hydrogen bonding of their bases; for example, A with T or U, and G with C.β conformation:（β构象）、An extended, zigzag arrangement of a polypeptide chain; a common secondary structure in proteins.βoxidation:（β氧化）Oxidative degradation of fatty acids into acetyl-CoA by successive oxidations at the β-carbon atom.β-turn:（β转角）A type of secondary structure in polypeptides consisting of four aminoacid residues arranged in a tight turn so that the polypeptide turns back on itself. Bilayer:（双分子层）A double layer of oriented amphipathic lipid molecules, forming the basic structure of biological membranes. The hydrocarbon tails face inward to form a continuous nonpolar phase.Bile salts:（胆酸盐）Amphipathic steroid derivatives with detergent properties, participating in digestion and absorption of lipids.Binding energy:（吸附能）The energy derived from noncovalent interactions between enzyme and substrate or receptor and ligand.Binding site:（结合部位）The crevice or pocket on a protein in which a ligand binds. Biocytin:（生物胞素）The conjugate amino acid residue arising from covalent attachment of biotin, through an amide linkage, to a Lys residue.Biomolecule:（生物分子）An organic compound normally present as an essential component of living organisms.Biopterin:（生物喋呤）An enzymatic cofactor derived from pterin and involved in certain oxidation-reduction reactions.Biosphere:（生物圈）All the living matter on or in the earth, the seas, and the atmosphere.Biotin:（生物素）A vitamin; an enzymatic cofactor involved in carboxylation reactions. Bond energy:（键能）The energy required to break a bond.Branch migration:（分支迁移）Movement of the branch point in branched DNA formed from two DNA molecules with identical sequences. See also Holliday intermediate.Buffer:（缓冲液）A system capable of resisting changes in pH, consisting of a conjugate acid-base pair in which the ratio of proton acceptor to proton donor is near unity.CCalorie:（卡）The amount of heat required to raise the temperature of 1.0 g of water from 14.5 to 15.5 °C. One calorie (cal) equals 4.18 joules (J).Calvin cycle:（Calvin循环）The cyclic pathway used by plants to fix carbon dioxide and produce triose phosphates.cAMP:See cyclic AMP.cAMP receptor protein (CRP): （cAMP受体蛋白）A specific regulatory protein that controls initiation of transcription of the genes producing the enzymes required for a bacterial cell to use some other nutrient when glucose is lacking. Also called catabolite gene activator protein (CAP)，降解物基因活化蛋白.CAP:See catabolite gene activator protein.Capsid:（衣壳）The protein coat of a virion or virus particle.Carbanion:（碳负离子）A negatively charged carbon atom.Carbocation: （碳正离子）A positively charged carbon atom; also called a carboniumion.Carbon-assimilation reactions:（碳同化反应）Reaction sequences in which atmospheric CO2 is converted into organic compounds.Carbon-fixation reaction:（固碳反应）The reaction catalyzed by rubisco during photosynthesis, or by other carboxylases, in which atmospheric CO2is initially incorporated into an organic compound.Carboxyl-terminal residue:（羧基末端残基）The only amino acid residue in a polypeptide chain with a free a-carboxyl group; defines the carboxyl terminus of the polypeptide.Carotenoids:（类葫罗卜素）Lipid-soluble photosynthetic pigments made up of isoprene units.Catabolism:（分解代谢）The phase of intermediary metabolism concerned with the energy-yielding degradation of nutrient molecules.Catabolite gene activator protein (CAP):See cAMP receptor protein.Catalytic site:（催化部位）See active site.Catecholamines:（儿茶酚胺类）Hormones, such as epinephrine, that are amino derivatives of catechol.Catenane:（连环体）Circular polymeric molecules with a noncovalent topological link resembling the links of a chain.Cation-exchange resin:（阳离子交换树脂）An insoluble polymer with fixed negative charges; used in the chromatographic separation of cationic substances.cDNA: See complementary DNA.Central dogma:（中心法则）The organizing principle of molecular biology: genetic information flows from DNA to RNA to protein.Centromere:（着丝粒） A specialized site within a chromosome, serving as the attachment point for the mitotic or meiotic spindle.Cerebroside(脑苷酯) Sphingolipid containing one sugar residue as a head group. Channeling:（生物合成途径限制作用)The direct transfer of a reaction product (common intermediate) from the active site of one enzyme to the active site of a different enzyme catalyzing the next step in a sequential pathway.Chemiosmotic coupling:（化学渗透偶联）Coupling of ATP synthesis to electron transfer via an electrochemical H+ gradient across a membrane.Chemotaxis（向化性）:A cell's sensing of and movement toward, or away from, a specific chemical agent.Chemotroph:（化能生物）An organism that obtains energy by metabolizing organic compounds derived from other organisms.Chiral center:（手性中心）An atom with substituents arranged so that the molecule is not superimposable on its mirror image.Chiral compound:（手性化合物）A compound that contains an asymmetric center(chiral atom or chiral center) and thus can occur in two nonsuperimposable mirror-image forms (enantiomers).Chlorophylls:（叶绿素）A family of green pigments functioning as receptors of light energy in photosynthesis; magnesium-porphyrin complexes.Chloroplasts:（叶绿体）Chlorophyll-containing photosynthetic organelles in some eukaryotic cells.Chromatin:（染色质）A filamentous complex of DNA, histones, and other proteins, constituting the eukaryotic chromosome.Chromatography:（层析）A process in which complex mixtures of molecules are separated by many repeated partitionings between a flowing (mobile) phase and a stationary phase.Chromosome:（染色体）A single large DNA molecule and its associated proteins, containing many genes; stores and transmits genetic information.Chylomicron:（乳糜微粒）A plasma lipoprotein consisting of a large droplet of triacylglycerols stabilized by a coat of protein and phospholipid; carries lipids from the intestine to the tissues.cis and trans isomers:（顺反异构体）See geometric isomers.Cistron:（顺反子）A unit of DNA or RNA corresponding to one gene.Citric acid cycle:（柠檬酸循环）A cyclic system of enzymatic. reactions for the oxidation of acetyl residues to carbon dioxide, in which formation of citrate is the first step; also known as the Krebs cycle or tricarboxylic acid cycle.Clones:（克隆）The descendants of a single cell.Cloning:The production of large numbers of identical DNA molecules, cells, or organisms, from a single ancestral DNA molecule, cell, or organism.Closed system:（封闭系统）A system that exchanges neither matter nor energy with the surroundings. See also system.Cobalamin:（钴胺素）See cocnzyme B12.Codon:（密码子）A sequence of three adjacent nucleotides in a nucleic acid that codes for a specific amino acid.Coenzyme:（辅酶）An organic cofactor required for the action of certain enzymes; often contains a vitamin as a component.Coenzyme A: （辅酶A）A pantothenic acid-containing coenzyme serving as an acyl group carrier in certain enzymatic reactions.Coenzyme B12: An enzymatic cofactor derived from the vitamin cobalamin, involved in certain types of carbon skeletal rearrangements.Cofactor(辅助因子) An inorganic ion or a coenzyme required for enzyme activity. Cognate:（相关的）Describing two biomolecules that normally interact; for example, an enzyme and its normal substrate, or a receptor and its normal ligand.Cohesive ends:（粘性末端）See sticky ends.Cointegrate:（共整合）An intermediate in the migration of certain DNA transposons in which the donor DNA and target DNA are covalently attached.Colligative properties:（依数性）Properties of solutions that depend on the number of solute particles per unit volume; for example, freezing-point depression.Common intermediate:（共同中间产物）A chemical compound common to two chemical reactions, as a product of one and a reactant in the other.Competitive inhibition:（竞争性抑制作用）A type of enzyme inhibition reversed by increasing the substrate concentration; a competitive inhibitor generally competes with the normal substrate or ligand for a protein's binding site.Complementary:（互补）Having a molecular surface with chemical groups arranged to interact specifically with chemical groups on another molecule.Complementary DNA (cDNA): A DNA used in DNA cloning, usually made by reverse transcriptase; complementary to a given mRNA.Configuration:（构型）The spatial arrangement of an organic molecule that is conferred by the presence of either (1) double bonds, about which there is no freedom of rotation, or (2) chiral centers, around which substituent groups are arranged in a specific sequence. Configurational isomers cannot be interconverted without breaking one or more covalent bonds.Conformation:（构象）The spatial arrangement, of substituent groups that are free to assume different positions in space, without breaking any bonds, because of the freedom of bond rotation.Conjugate acid-base pair:（共扼酸碱对） A proton donor and its corresponding deprotonated species; for example, acetic acid (donor) and acetate (acceptor). Conjugate redox pair: （共扼氧还对）An electron donor and its corresponding electron acceptor form; for example, Cu+ (donor) and Cu2+ (acceptor), or NADH (donor) and NAD+ (acceptor).Conjugated protein:（结合蛋白质）A protein containing one or more prosthetic groups.Consensus sequence:（一致序列）A DNA or amino acid sequence consisting of the residues that occur most commonly at each position within a set of similar sequences. Conservative substitution:（保守性置换）Replacement of an amino acid residue in a polypeptide by another residue with similar properties; for example, substitution of Glu by Asp.Constitutive enzymes:（组成酶）Enzymes required at all times by a cell and present at some constant level; for example, many enzymes of the central metabolic pathways. Sometimes called house-keeping enzymes.Contour length (外形长度): The length of a helical polymeric molecule as measured along the molecule's helical axis.Corticosteroids（皮质类固醇激素）Steroid hormones formed by the adrenal cortex. Cotransport:（共转运）The simultaneous transport, by a single transporter, of twosolutes across a membrane. See antiport, symport.Coupled reactions:（偶联反应）Two chemical reactions that have a common intermediate and thus a means of energy transfer from one to the other.Covalent bond:（共价键）A chemical bond that involves sharing of electron pairs. Cristae:（嵴）Infoldings of the inner mitochondrial membrane.CRP（cAMP受体蛋白）See cAMP receptor protein.Cyclic AMP (cAMP):A second messenger within cells; its formation by adenylyl cyclase is stimulated by certain hormones or other molecular signals.Cyclic electron flow:（循环电子流）In chloroplasts, the light-induced flow of electrons originating from and returning to photosystem I.Cyclic photophosphorylation: （循环光合磷酸化）ATP synthesis driven by cyclic electron flow through photosystem I.Cyclin:（细胞周期蛋白）One of a family of proteins that activate cyclin-dependent protein kinases and thereby regulate the cell cycle.Cytochromes:（细胞色素）Heme proteins serving as electron carriers in respiration, photosynthesis, and other oxidation-reduction reactions.Cytokine:（细胞因子）One of a family of small secreted proteins (such as interleukins or interferons) that activate cell division or differentiation by binding to plasma membrane receptors in sensitive cells.Cytokinesis:（胞质分裂）The final separation of daughter cells following mitosis. Cytoplasm:（细胞质）The portion of a cell's contents outside the nucleus but within the plasma membrane; includes organelles such as mitochondria.Cytoskeleton:（细胞骨架）The filamentous network providing structure and organization to the cytoplasm; includes actin filaments, microtubules, and intermediate filaments.Cytosol:（细胞浆）The continuous aqueous phase of the cytoplasm, with its dissolved solutes; excludes the organelles such as mitochondria.DDalton:（道尔顿）The weight of a single hydrogen atom (1.66 x I0-24 g).Dark reactions:（暗反应）See carbon-assimilation reactions.De novo pathway:（从头合成）Pathway for synthesis of a biomolecule, such as a nucleotide, from simple precursors; as distinct from a salvage pathway. Deamination:（脱氨基作用）The enzymatic removal of amino groups from biomolecules such as amino acids or nucleotides.Degenerate code:（兼并密码）A code in which a single element in one language is specified by more than one element in a second language.Dehydrogenases:（脱氢酶类）Enzymes catalyzing the removal of pairs of hydrogen atoms from their substrates.Deletion mutation:（删除突变）A mutation resulting from the deletion of one or morenucleotides from a gene or chromosome.Denaturation:（变性）Partial or complete unfolding of the specific native conformation of a polypeptide chain, protein, or nucleic acid.Denatured protein:（变性蛋白）A protein that has lost its native conformation by exposure to a destabilizing agent such as heat or detergent.Deoxyribonucleic acid; See DNA.Deoxyribonucleotides:（脱氧核糖核苷酸）Nucleotides containing 2-deoxyribose as the pentose component.Desaturases:（去饱和酶）Enzymes that catalyze the introduction of double bonds into the hydrocarbon portion of fatty acids.Desolvation:（脱水）In aqueous solution, the release of bound water surrounding a solute.Dextrorotatory isomer:9右旋异构体） A stercoisomer that rotates the plane of plane-polarized light clockwise.Diabetes mellitus:（糖尿病）A metabolic disease resulting from insulin deficiency; characterized by a failure in glucose transport from the blood into cells at normal glucose concentrations.Dialysis:（透析）Removal of small molecules from a solution of a macromolecule, by allowing them to diffuse through a semipermeable membrane into water.Differential centrifugation:（差速离心）Separation of cell organelles or other particles of different size by their different rates of sedimentation in a centrifugal field. Differentiation:（分化）Specialization of cell structure and function during embryonic growth and development.Diffusion:（扩散）The net movement, of molecules in the direction of lower concentration.Digestion:（消化）Enzymatic hydrolysis of major nutrients in the gastrointestinal system to yield their simpler components.Diploid:（二倍体）Having two sets of genetic information; describing a cell with two chromosomes of each type.Dipole;（双极分子）A molecule having both positive and negative charges.Diprotic acid: An acid having two dissociable protons.Disaccharide:（二糖） A carbohydrate consisting of two covalently joined monosaccharide units.Dissociation constant:（解离常数）(1) An equilibrium constant (K d)for the dissociation of a complex of two or more biomolecules into its components; for example, dissociation of a substrate from an enzyme. (2) The dissociation constant (Ka) of an acid, describing its dissociation into its conjugate base and a proton.Disulfide bridge:(二硫桥)A covalent cross link between two polypeptide chains formed by a cystine residue (two Cys residues).DNA (deoxyribonucleic acid)： A polynucleotide having a specific sequence of deoxyribonucleotide units covalently joined through 3', 5'-phosphodiester bonds; serves as the carrier of genetic information.DNA chimera:（DNA嵌合）A DNA containing genetic information derived from two different species.DNA cloning:Sec cloning.DNA library:（DNA文库）A collection of cloned DNA fragments.DNA ligase：（DNA连接酶）An enzyme that creates a phosphodiester bond between the 3' end of one DNA segment, and the 5' end of another.DNA looping:（DNA出环）The interaction of proteins bound at distant sites on a DNA molecule so that the intervening DNA forms a loop.DNA microarray:（DNA微阵列）A collection of DNA sequences immobilized on a solid surface, with individual sequences laid out in patterned arrays that can be probed by hybridization.DNA polymerase:（DNA聚合酶）An enzyme that catalyzes template-dependent synthesis of DNA from its deoxyribonucleoside 5'-triphosphate precursors.DNA replicase system：（DNA复制酶系统）The entire complex of enzymeH and specialized proteins required in biological DNA replication.DNA supercoiling:（DNA超螺旋化）The coiling of DNA upon itself, generally as a result of bending, underwinding, or overwinding of the DNA helix.DNA transposition:（DNA转座）See transposition.domain:（结构域）A distinct structural unit of a polypeptide; domains may have separate functions and may fold as independent, compact units.Double helix:（双螺旋）The natural coiled conformation of two complementary, antiparallel DNA chains.Double-reciprocal plot:（双倒数作图）A plot, of 1/Vo versus 1/[S], which allows a more accurate determination of Vmax and Km than a plot of V versus [S]; also called the Lineweaver-Burk plot,EE'°: 标准还原电位See standard reduction potential.E. coli (Escherichia coli):（大肠杆菌）A common bacterium found in the small intestine of vertebrates; the most well-studied organism.Electrochemical gradient:（电化学梯度）The sum of the gradients of concentration and of electric charge of an ion across a membrane; the driving force for oxidative phosphorylation and photophosphorylation.Electrochemical potential: （电化学势）The energy required to maintain a separation of charge and of concentration across a membrane.Electrogenic:（生电的）Contributing to an electrical potential across a membrane.Electron acceptor:（电子受体） A substance that receives electrons in an oxidation-reduction reaction.Electron carrier:（电子载体）A protein, such as a flavoprotein or a cytochrome, that can reversibly gain and lose electrons; functions in the transfer of electrons from organic nutrients to oxygen or some other terminal acceptor.Electron donor:（电子供体） A substance that donates electrons in an oxidation-reduction reaction.Electron transfer:（电子转移）Movement of electrons from substrates to oxygen via the carriers of the respiratory (electron transfer) chain.Electrophile:（亲电剂）An electron-deficient group with a strong tendency to accept electrons from an electron-rich group (nucleophile).Electrophoresis（电泳）: Movement of charged solutes in response to an electrical field; often used to separate mixtures of ions, proteins, or nucleic acids.Electroporation:（电穿孔法）Introduction of macromolecules into cells after rendering the cells transiently permeable by the application of a high-voltage pulse.Elongation factors:（延长因子）Specific proteins required in the elongation of polypeptide chains by ribosomes.Eluate:（流出液）The effluent from a chromatographic column.Enantiomers:（对映异构体）Stereoisomers that are nonsuperimposable mirror images of each other.End-product inhibition:See feedback inhibition.Endergonic reaction(耗能反应): A chemical reaction that consumes energy (that is, for which ΔG is positive).Endocrine glands:（内分泌腺）Groups of cells specialized to synthesize hormones and secrete them into the blood to regulate other types of cells.Endocytosis:（内吞体）The uptake of extracellular material by its inclusion within a vesicle formed by an invagination of the plasma membrane.Endonuclease:（内切核酸酶）An enzyme that hydrolyzes the interior phosphodiester bonds of a nucleic acid; that is, it acts at points other than the terminal bonds. Endoplasmic reticulum:（内质网）An extensive system of double membranes in the cytoplasm of eukaryotic cells; it encloses secretory channels and is often studded with ribosomes (rough endoplasmic reticulum).Endothermic reaction:（吸热反应）A chemical reaction that takes up heat (that is, for which ΔH is positive).Energy charge:（能荷）The fractional degree to which the ATP/ADP/AMP system is filled with high-energy phosphate groups.Energy coupling:（能量偶联）The transfer of energy from one process to anotlier. Enhancers:（增强子）DNA sequences that facilitate the expression of a given gene;。

三氟乙酸脱苄基O-debenzylation_of_ortho-substituted_phenols_with_trifluoroacetic_acid

Mild,efﬁcient and rapid O-debenzylation of ortho -substituted phenols with triﬂuoroacetic acidSteven Fletcher *,Patrick T.Gunning *Department of Chemistry,University of Toronto,Mississauga,ON L5L 1C6,Canadaa r t i c l e i n f o Article history:Received 21May 2008Revised 2June 2008Accepted 4June 2008Available online 10June 2008a b s t r a c tThe mild and efﬁcient deblocking of aryl benzyl ethers with TFA is reported.Cleavage was fastest with ortho -electron-withdrawing groups on the phenolic ring,which we have attributed to a proton chelation effect,furnishing the deprotected phenols in excellent yields.The corresponding para -methoxybenzyl,allyl and iso -propyl ethers were also cleanly removed under these conditions.In addition,the selective aryl benzyl ether debenzylation in the presence of benzyl ester,Cbz carbamate and Boc carbamate func-tionalities was also observed.Crown Copyright Ó2008Published by Elsevier Ltd.All rights reserved.Phosphotyrosines feature in the design of inhibitors of several protein targets,including protein tyrosine phosphatase 1B (PTP1B).1However,these moieties suffer from hydrolytic lability to cellular phosphatases and poor cell penetration due to the asso-ciated dianionic charge.1To address these issues,salicylic acid derivatives (and closely-related analogues)have become popular mimetics of phosphotyrosine in small molecule inhibitors.1–5Turk-son et al.have recently reported on NSC74859(1),a potent,sali-cylic acid-based inhibitor of the oncogenic protein Stat3.6As part of our structure–activity relationship (SAR)studies on NSC74859(1),we sought to debenzylate both the phenol ether and benzoate ester in 2without reducing the aryl-bromide bond,a common undesired side reaction that occurs with hydrogen gas and Pd/C catalyst.7O -Benzyl-protected phenols are known to undergo debenzyla-tion with triﬂuoroacetic acid (TFA)8by an initial protonation of the weakly basic phenol oxygen,although additives such as strongorganic acids (e.g.,triﬂuoromethanesulfonic acid 9)or a large excess of nucleophilic scavenger (e.g.,thioanisole,which accelerates the reaction by a ‘push–pull’mechanism 10)are typically required.Re-cent work by Ploypradith et al.describes the mild deprotection of aromatic ethers with sub-stoichiometric para -toluenesulfonic acid on solid support.11In a special case,O -benzyl-protected ortho -nitrophenol was cleaved rapidly (<5min)with neat TFA,12which we considered was due to the ability of the substrate to chelate a proton since the structurally-similar ortho -hydroxybenzoates (salicylates)are well-known to chelate copper ions and iron ions.We reasoned that 2(and indeed 3)may similarly undergo acceler-ated debenzylation with TFA.In fact,as shown in Scheme 1,treat-ment of 2(or 3)with a 1:1mixture of TFA/toluene led to rapid debenzylation (5min for 2;1h for 3)in 91%yield for 2(or 85%yield for 3).In this Letter,we will explore the structural require-ments of the phenol component that increase the lability of the O -benzyl phenol ether bond in the presence of TFA.In addition,0040-4039/$-see front matter Crown Copyright Ó2008Published by Elsevier Ltd.All rights reserved.*Tel.:+19058285354;fax:+19058285425(P.T.G.).E-mail addresses:steven.ﬂetcher@utoronto.ca (S.Fletcher),patrick.gunning@utoronto.ca (P.T.Gunning).Tetrahedron Letters 49(2008)4817–4819Contents lists available at ScienceDirectTetrahedron Lettersj o ur na l h om e pa ge :w w w.e ls e v ie r.c o m/lo c at e/t et l e twe will explore the selectivity of this mild debenzylation tech-nique with respect to other aromatic ethers and examine the sta-bility of other benzyl-based protecting groups to these reaction conditions.A series of 12O -benzyl-protected phenols was prepared by standard procedures in near quantitative yields.Each of these ethers was then deprotected with a 1:1mixture of TFA/toluene;our observations are summarized in Table 1.In certain cases,O ?C benzyl migration (Friedel–Crafts reaction)by-products (610%)were occasionally inseparable from the product by silica gel ﬂash column chromatography.Thus,several benzyl cation cap-tors were investigated for their abilities to improve yields and puri-ties of the debenzylation reactions.Three to ten equivalents of p -cresol,anisole and triethylsilane were employed,but these exerted little effects on reducing by-product formation.Conversely,we dis-covered that including the more nucleophilic scavenger thioanisole as an additive to the co-solvent toluene typically,after silica gel ﬂash column chromatography,furnished products in P 95%puri-ties (and higher yields),as judged by 1H NMR.Nevertheless,we envisaged any Friedel–Crafts impurities would be more readily separable on slightly more complex aryl benzyl ethers,as we ob-served with the substrates shown in Scheme 1and Tables 3and 4(>99%purities (1H NMR)in each case).Whilst likely leading to even higher yields and purities,large excesses of thioanisole (50equiv)are also known to accelerate TFA-mediated debenzyla-tion.10However,in our hands just 3equiv of thioanisole had little effect on the rate of debenzylation,allowing us to attribute the deprotection rates solely to the structure of the phenol.Electron-rich phenols are good scavengers of benzyl cations,13and since preliminary experiments with electron-rich phenols generated complex mixtures of Friedel–Crafts by-products under these deb-enzylation conditions,we chose to investigate only electron-poor phenols in this study.O -Benzyl-protected phenols with p -ortho -electron-withdraw-ing groups (6a ,6b ,6d ,6f )were swiftly (several in less than 3h cf.24h for unsubstituted phenol 6l )and cleanly debenzylated,with less than 5%of the undesired C-benzylated phenol by-prod-ucts.In contrast,meta -and para -electron-withdrawing groups slo-wed down the debenzylation (e.g.,entries 6g and 6h ),relative to the control compound 6l ,which itself could only be obtained in moderate purity by this method.The r -withdrawing (and p -donating)bromophenols 6i –k were insufﬁciently deactivated to benzyl cation scavenging and were contaminated with several by-products.Importantly,n -butyl benzyl ether 8was unaffected by TFA under the reaction conditions,indicating this procedure is selective for aryl benzyl ethers.In addition,the results in Table 1suggest that this procedure is suitable only for phenols substituted with p -electron-withdrawing groups.Since the debenzylation mechanism with TFA proceeds via an initial protonation of the phenol ether oxygen,the more available the ether oxygen lone pairs are,the faster the reaction will be.Hence,the slower reaction times for the phenols bearing meta -and para -electron-withdrawing groups make sense,although this is not true for the ortho -functionalized aryl benzyl ethers.As hypothesized for the bis-benzyl salicylate derivative 2earlier,we considered these ortho -substituted phenols were capable of chelat-ing the acidic hydrogen atom from TFA which therein facilitated the acid-mediated debenzylation via a six-membered cyclic inter-mediate,as proposed in Scheme 2.A similar chelation intermediate has been put forward by Baldwin and Haraldsson to account for the Lewis acid MgBr 2-mediated debenzylation of aromatic benzyl ethers ortho to an aldehyde group.14Accordingly,to test this hypothesis we expanded this series of ortho -substituted aryl benzyl ethers,and the results from their deb-enzylation reactions with TFA are summarized in Table 2.These substrates have been listed in order of increasing approximateTable 1TFA-mediated debenzylation of O -benzyl-protected phenols aTFAtolueneOBnROHR67Substrate RTime (h)b Yield c (%)6a o -CO 2Me,m d -NHAc 5min 936b o -CO 2Me 5min 946c p -CO 2Me 36e 63(85f )6d o -CO 2Bn 5min 936e p -CO 2Bn 36e 58(79f )6f o -NO 23976g m -NO 236e 75(98f )6h p -NO 236e 66(98f )6i o -Br 16—g 6j m -Br 30—g 6k p -Br 36—g 6lH 24—gn -BuOBn (8)—24No reactionaThe reaction was carried out with 6(0.5mmol)in a 1:1mixture of TFA/toluene (5ml)at rt,with 3equiv of thioanisole.bTime taken for all starting material to be consumed.cIsolated yield after silica gel ﬂash column chromatography.dmeta to phenol oxygen AND para to ester.eReaction was slow and incomplete after 3days.fYield based on recovered starting material.gComplex mixture of products.Table 2TFA-mediated debenzylation of O -benzyl-protected,ortho -substituted phenols aTFA tolueneOBnOH67RRSubstrate R p K aH b Time c (h)Yield d (%)Relative rate 6m CO 2NH 2À2248316n CHO À7 3.594e 6.96o CO 2H À8191246b CO 2Me À8.55min 942886d CO 2Bn À8.55min 932886p CN À10>4851(95f )—6f NO 2À1239786i Br —16—g 1.56lH—24—g1aThe reaction was carried out with 6(0.5mmol)in a 1:1mixture of TFA/toluene (5ml)at rt,with 3equiv of thioanisole.bApproximate p K aH of conjugate acid of R group.15cTime taken for all starting material to be consumed.dIsolated yield after silica gel ﬂash column chromatography.eIncluding thioanisole in the deprotection of 6n led to further by-products,thus no scavenger was used and compound 7n could be obtained in only 90%purity.fYield based on recovered starting material.gComplex mixture of products.4818S.Fletcher,P.T.Gunning /Tetrahedron Letters 49(2008)4817–4819acidity of the conjugate acid (decreasing p K aH )of the ortho -elec-tron-withdrawing substituent.15There appears to be an optimal p K aH of around À8.5,that is exhibited by carboxylic esters,which lead to the fastest rate of debenzylation with TFA.In an approxi-mate bell-shaped distribution of reaction rate versus ortho -substi-tuent p K aH —that was interrupted only by ortho -cyanophenol 6p —protonatable groups with p K aH ’s <À8.5or >À8.5were less effective at accelerating the TFA-mediated debenzylation.These data concur with our chelation hypothesis:groups that are too ba-sic bind more strongly to the TFA proton making it less available for sharing with,and ultimately releasing to,the phenol ether oxygen;groups that are weakly basic do not bind the TFA proton as well,leading to reduced chelation and hence less rate enhancement.The anomalous result for ortho -cyanophenol 6p was anticipated since this compound was selected as a negative control.Phenol 6p is geometrically incapable of chelating a proton,because the lin-ear,sp -hybridized nitrile functionality directs its basic nitrogen atom (p K aH %À10)away from the phenol oxygen.As predicted,there was no rate enhancement for the TFA-mediated debenzyla-tion of 6p relative to phenol 6l .In fact,6p was only slowly deben-zylated,at a rate that was comparable with the m -nitro and p -nitro derivatives 6g and 6h ,respectively.We next wanted to investigate the selectivity for the deprotec-tion of the benzyl group over other phenol protecting groups.Accordingly,the benzyl group in salicylate derivative 9a was varied with para -methoxybenzyl (PMB;9b ),methyl (9c ),allyl (9d )and iso -propyl (i -Pr;9e ).These substrates were then debenzylated with a 1:1mixture of TFA/toluene;our ﬁndings are reported in Table 3.Any impurities this time were minor and readily separable from the products,eliminating the need for the additive thioanisole.The relative rates at which these protecting groups were removed was para -methoxybenzyl >benzyl >allyl >iso -propyl )methyl,which reﬂects the stability of the carbocations.These data suggest that in salicylates such as 9,the benzyl phenol protecting group (R =Bn)can be removed with TFA in the presence of the corres-ponding allyl,iso -propyl and methyl ethers.Finally,we explored the selectivity of this mild debenzylation technique over other benzyl-based protecting groups,as shown in Table 4.As the results demonstrate,it was possible to deblock the O -benzyl ether in the presence of a benzyl ester (6d )and in the presence of a benzyl carbamate (11b ),thereby increasing the orthogonality of O -benzyl phenol ethers of salicylate derivatives.Interestingly,it was even possible to cleave the benzyl group in 11c with TFA in the presence of an N -Boc-protected aniline.In summary,we have presented the mild,efﬁcient and rapid deblocking of ortho -substituted aryl benzyl ethers with TFA.Deb-enzylation was fastest when the ortho group was a carboxylic ester,which we have attributed to a proton chelation effect.Other ortho groups that accelerated the TFA-mediated debenzylation included carboxylic acid,aldehyde and nitro.In addition,we have shown that in such ortho -functionalized phenols,benzyl could be removed in the presence of the corresponding iso -propyl,allyl and methyl ethers.Moreover,the benzyl ether could be selectively cleaved in the presence of benzyl ester,Cbz carbamate and Boc carbamate functionalities.AcknowledgementsThe authors gratefully acknowledge ﬁnancial support for this work from the Canadian Foundation of Innovation and the Univer-sity of Toronto (Connaught Foundation).References and notes1.Zhang,S.;Zhang,Z.-Y.Drug Discov.Today 2007,12,373–381.2.(a)Pei,Z.;Li,X.;Liu,G.;Abad-Zapatero,C.;Lubben,T.;Zhang,T.;Ballaron,S.J.;Hutchins,C.W.;Trevillyana,J.M.;Jirouseka,M.R.Bioorg.Med.Chem.Lett.2003,13,3129–3132;(b)Xin,Z.;Liu,G.;Abad-Zapatero,C.;Pei,Z.;Szczepankiewicz,B.G.;Li,X.;Zhang,T.;Hutchins,C.W.;Hajduk,P.J.;Ballaron,S.J.;Stashko,M.A.;Lubben,T.H.;Trevillyana,J.M.;Jirouseka,M.R.Bioorg.Med.Chem.Lett.2003,13,3947–3950.3.Tautz,L.;Bruckner,S.;Sareth,S.;Alonso,A.;Bogetz,J.;Bottini,N.;Pellecchia,M.;Mustelin,T.J.Biol.Chem.2005,280,9400–9408.4.Shrestha,S.;Bhattarai,B.R.;Chang,K.J.;Leea,K.-H.;Choa,H.Bioorg.Med.Chem.Lett.2007,17,2760–2764.5.Liljebris,C.;Larsen,S.D.;Ogg,D.;Palazuk,B.J.;Bleasdale,J.E.J.Med.Chem.2002,45,1785–1798.6.Siddiquee,K.;Zhang,S.;Guida,W.C.;Blaskovich,M.A.;Greedy,B.;Lawrence,H.R.;Yip,M.L.R.;Jove,R.;Laughlin,M.M.;Lawrence,N.J.;Sebti,S.M.;Turkson,J.Proc.Natl.Acad.Sci.U.S.A.2007,104,7391–7396.7.Pandey,P.N.;Purkayastha,M.L.Synthesis 1982,876–878.8.(a)Greene,T.W.;Wuts,P.G.M.Protective Groups in Organic Synthesis ,3rd ed.;John Wiley &Sons:New York,1999;(b)Kocienski,P.J.Protecting Groups ,3rd ed.;Georg Thieme:Stuttgart,Germany,2003.9.Kiso,Y.;Isawa,H.;Kitagawa,K.;Akita,T.Chem.Pharm.Bull.1978,26,2562–2564.10.Kiso,Y.;Ukawa,K.;Nakamura,S.;Ito,K.;Akita,T.Chem.Pharm.Bull.1980,28,673–676.11.Ploypradith,P.;Cheryklin,P.;Niyomtham,N.;Bertoni,D.R.;Ruchirawat,.Lett.2007,9,2637–2640.12.Marsh,J.P.,Jr.;Goodman,.Chem.1965,30,2491–2492.13.(a)Eberle,A.N.J.Chem.Soc.,Perkin Trans.11986,361–367;(b)Bodanszky,M.;Tolle,J.C.;Deshmane,S.S.;Bodanszky,A.Int.J.Pept.Protein Res.1978,12,57–68.14.Haraldsson,G.G.;Baldwin,J.E.Tetrahedron 1997,53,215–224.15.(a)Ionization Constants of Organic Acids in Solution ;Serjeant,E.P.,Dempsey,B.,Eds.IUPAC Chemical Data Series No.23;Pergamon Press:Oxford,UK,1979;(b)see also:/labs/evans/pdf/evans_pKa_table.pdf .Table 3TFA-mediated deprotection of O-blocked phenol ether derivatives of methyl 4-acetamidosalicylate aTFAtolueneNHAcNHAcORO OMeOH OMeO 910Substrate R Time b (h)Yield c (%)9a Bn 5min 919b PMB 2min 909c Me 480d 9d Allyl 20919ei -Pr3692aThe reaction was carried out with 9(0.5mmol)in a 1:1mixture of TFA/toluene (5ml)at rt.bTime taken for all starting material to be consumed.cIsolated yield after silica gel ﬂash column chromatography.dOnly starting material remained after 48h,at which point the reaction was aborted.Table 4Selectivity investigation into the TFA-mediated debenzylation of aryl benzyl ethers aTFA tolueneOBnOH2Bn2Bn1112RRSubstrate R Yield b (%)6d c H 9311a NHAc 9211b NHCbz 9311c dNHBoc54aThe reaction was carried out with 11(0.5mmol)in a 1:1mixture of TFA/toluene (5ml)at rt for 5min,then all solvents were evaporated.bIsolated yield after silica gel ﬂash column chromatography.cFor compound 6d ,3equiv of thioanisole were also used.dAfter 5min,the reaction mixture was diluted with CH 2Cl 2and then immedi-ately neutralized with 1M NaOH.The organic layer was then separated and evaporated.S.Fletcher,P.T.Gunning /Tetrahedron Letters 49(2008)4817–48194819。

药物分析专业英语词汇

药物分析专业英语词汇AAbbe refractometer 阿贝折射仪absorbance 吸收度absorbance ratio 吸收度比值absorption 吸收absorption curve 吸收曲线absorption spectrum 吸收光谱absorptivity 吸收系数accuracy 准确度acid-dye colorimetry 酸性染料比色法acidimetry 酸量法acid-insoluble ash 酸不溶性灰分acidity 酸度activity 活度additive 添加剂additivity 加和性adjusted retention time 调整保留时间adsorbent 吸附剂adsorption 吸附affinity chromatography 亲和色谱法aliquot （一）份alkalinity 碱度alumina 氧化铝ambient temperature 室温ammonium thiocyanate 硫氰酸铵analytical quality control（AQC）分析质量控制anhydrous substance 干燥品anionic surfactant titration 阴离子表面活性剂滴定法antibiotics-microbial test 抗生素微生物检定法antioxidant 抗氧剂appendix 附录application of sample 点样area normalization method 面积归一化法argentimetry 银量法arsenic 砷arsenic stain 砷斑ascending development 上行展开ash-free filter paper 无灰滤纸（定量滤纸）assay 含量测定assay tolerance 含量限度atmospheric pressure ionization(API) 大气压离子化attenuation 衰减Bback extraction 反萃取back titration 回滴法bacterial endotoxins test 细菌内毒素检查法band absorption 谱带吸收baseline correction 基线校正baseline drift 基线漂移batch, lot 批batch(lot) number 批号Benttendorff method 白田道夫（检砷）法between day (day to day, inter-day) precision 日间精密度between run (inter-run) precision 批间精密度biotransformation 生物转化bioavailability test 生物利用度试验bioequivalence test 生物等效试验biopharmaceutical analysis 体内药物分析，生物药物分析blank test 空白试验boiling range 沸程British Pharmacopeia (BP) 英国药典bromate titration 溴酸盐滴定法bromimetry 溴量法bromocresol green 溴甲酚绿bromocresol purple 溴甲酚紫bromophenol blue 溴酚蓝bromothymol blue 溴麝香草酚蓝bulk drug, pharmaceutical product 原料药buret 滴定管by-product 副产物Ccalibration curve 校正曲线calomel electrode 甘汞电极calorimetry 量热分析capacity factor 容量因子capillary zone electrophoresis (CZE) 毛细管区带电泳capillary gas chromatography 毛细管气相色谱法carrier gas 载气cation-exchange resin 阳离子交换树脂ceri(o)metry 铈量法characteristics, description 性状check valve 单向阀chemical shift 化学位移chelate compound 鳌合物chemically bonded phase 化学键合相chemical equivalent 化学当量Chinese Pharmacopeia (ChP) 中国药典Chinese material medicine 中成药Chinese materia medica 中药学Chinese materia medica preparation 中药制剂Chinese Pharmaceutical Association (CPA) 中国药学会chiral 手性的chiral stationary phase (CSP) 手性固定相chiral separation 手性分离chirality 手性chiral carbon atom 手性碳原子chromatogram 色谱图chromatography 色谱法chromatographic column 色谱柱chromatographic condition 色谱条件chromatographic data processor 色谱数据处理机chromatographic work station 色谱工作站clarity 澄清度clathrate, inclusion compound 包合物clearance 清除率clinical pharmacy 临床药学coefficient of distribution 分配系数coefficient of variation 变异系数color change interval （指示剂）变色范围color reaction 显色反应colorimetric analysis 比色分析colorimetry 比色法column capacity 柱容量column dead volume 柱死体积column efficiency 柱效column interstitial volume 柱隙体积column outlet pressure 柱出口压column temperature 柱温column pressure 柱压column volume 柱体积column overload 柱超载column switching 柱切换committee of drug evaluation 药品审评委员会comparative test 比较试验completeness of solution 溶液的澄清度compound medicines 复方药computer-aided pharmaceutical analysis 计算机辅助药物分析concentration-time curve 浓度－时间曲线confidence interval 置信区间confidence level 置信水平confidence limit 置信限congealing point 凝点congo red 刚果红（指示剂）content uniformity 装量差异controlled trial 对照试验correlation coefficient 相关系数contrast test 对照试验counter ion 反离子（平衡离子）cresol red 甲酚红（指示剂）crucible 坩埚crude drug 生药crystal violet 结晶紫（指示剂）cuvette, cell 比色池cyanide 氰化物cyclodextrin 环糊精cylinder, graduate cylinder, measuring cylinder 量筒cylinder-plate assay 管碟测定法Ddaughter ion （质谱）子离子dead space 死体积dead-stop titration 永停滴定法dead time 死时间decolorization 脱色decomposition point 分解点deflection 偏差deflection point 拐点degassing 脱气deionized water 去离子水deliquescence 潮解depressor substances test 降压物质检查法derivative spectrophotometry 导数分光光度法derivatization 衍生化descending development 下行展开desiccant 干燥剂detection 检查detector 检测器developer, developing reagent 展开剂developing chamber 展开室deviation 偏差dextrose 右旋糖，葡萄糖diastereoisomer 非对映异构体diazotization 重氮化2,6-dichlorindophenol titration 2,6-二氯靛酚滴定法differential scanning calorimetry (DSC) 差示扫描热量法differential spectrophotometry 差示分光光度法differential thermal analysis (DTA) 差示热分析differentiating solvent 区分性溶剂diffusion 扩散digestion 消化diphastic titration 双相滴定disintegration test 崩解试验dispersion 分散度dissolubility 溶解度dissolution test 溶出度检查distilling range 馏程distribution chromatography 分配色谱distribution coefficient 分配系数dose 剂量drug control institutions 药检机构drug quality control 药品质量控制drug release 药物释放度drug standard 药品标准drying to constant weight 干燥至恒重dual wavelength spectrophotometry 双波长分光光度法duplicate test 重复试验Eeffective constituent 有效成分effective plate number 有效板数efficiency of column 柱效electron capture detector 电子捕获检测器electron impact ionization 电子轰击离子化electrophoresis 电泳electrospray interface 电喷雾接口electromigration injection 电迁移进样elimination 消除eluate 洗脱液elution 洗脱emission spectrochemical analysis 发射光谱分析enantiomer 对映体end absorption 末端吸收end point correction 终点校正endogenous substances 内源性物质enzyme immunoassay(EIA) 酶免疫分析enzyme drug 酶类药物enzyme induction 酶诱导enzyme inhibition 酶抑制eosin sodium 曙红钠（指示剂）epimer 差向异构体equilibrium constant 平衡常数equivalence point 等当点error in volumetric analysis 容量分析误差excitation spectrum 激发光谱exclusion chromatography 排阻色谱法expiration date 失效期external standard method 外标法extract 提取物extraction gravimetry 提取重量法extraction titration 提取容量法extrapolated method 外插法，外推法Ffactor 系数，因数，因子feature 特征Fehling‘s reaction 费林反应field disorption ionization 场解吸离子化field ionization 场致离子化filter 过滤，滤光片filtration 过滤fineness of the particles 颗粒细度flame ionization detector(FID) 火焰离子化检测器flame emission spectrum 火焰发射光谱flask 烧瓶flow cell 流通池flow injection analysis 流动注射分析flow rate 流速fluorescamine 荧胺fluorescence immunoassay(FIA) 荧光免疫分析fluorescence polarization immunoassay(FPIA) 荧光偏振免疫分析fluorescent agent 荧光剂fluorescence spectrophotometry 荧光分光光度法fluorescence detection 荧光检测器fluorimetyr 荧光分析法foreign odor 异臭foreign pigment 有色杂质formulary 处方集fraction 馏分freezing test 结冻试验funnel 漏斗fused peaks, overlapped peaks 重叠峰fused silica 熔融石英Ggas chromatography(GC) 气相色谱法gas-liquid chromatography(GLC) 气液色谱法gas purifier 气体净化器gel filtration chromatography 凝胶过滤色谱法gel permeation chromatography 凝胶渗透色谱法general identification test 一般鉴别试验general notices （药典）凡例general requirements （药典）通则good clinical practices(GCP) 药品临床管理规范good laboratory practices(GLP) 药品实验室管理规范good manufacturing practices(GMP) 药品生产质量管理规范good supply practices(GSP) 药品供应管理规范gradient elution 梯度洗脱grating 光栅gravimetric method 重量法Gutzeit test 古蔡（检砷）法Hhalf peak width 半峰宽[halide] disk method, wafer method, pellet method 压片法head-space concentrating injector 顶空浓缩进样器heavy metal 重金属heat conductivity 热导率height equivalent to a theoretical plate 理论塔板高度height of an effective plate 有效塔板高度high-performance liquid chromatography (HPLC) 高效液相色谱法high-performance thin-layer chromatography (HPTLC) 高效薄层色谱法hydrate 水合物hydrolysis 水解hydrophilicity 亲水性hydrophobicity 疏水性hydroscopic 吸湿的hydroxyl value 羟值hyperchromic effect 浓色效应hypochromic effect 淡色效应Iidentification 鉴别ignition to constant weight 灼烧至恒重immobile phase 固定相immunoassay 免疫测定impurity 杂质inactivation 失活index 索引indicator 指示剂indicator electrode 指示电极inhibitor 抑制剂injecting septum 进样隔膜胶垫injection valve 进样阀instrumental analysis 仪器分析insulin assay 胰岛素生物检定法integrator 积分仪intercept 截距interface 接口interference filter 干涉滤光片intermediate 中间体internal standard substance 内标物质international unit(IU) 国际单位in vitro 体外in vivo 体内iodide 碘化物iodoform reaction 碘仿反应iodometry 碘量法ion-exchange cellulose 离子交换纤维素ion pair chromatography 离子对色谱ion suppression 离子抑制ionic strength 离子强度ion-pairing agent 离子对试剂ionization 电离，离子化ionization region 离子化区irreversible indicator 不可逆指示剂irreversible potential 不可逆电位isoabsorptive point 等吸收点isocratic elution 等溶剂组成洗脱isoelectric point 等电点isoosmotic solution 等渗溶液isotherm 等温线KKarl Fischer titration 卡尔·费歇尔滴定kinematic viscosity 运动黏度Kjeldahl method for nitrogen 凯氏定氮法Kober reagent 科伯试剂Kovats retention index 科瓦茨保留指数Llabelled amount 标示量leading peak 前延峰least square method 最小二乘法leveling effect 均化效应licensed pharmacist 执业药师limit control 限量控制limit of detection(LOD) 检测限limit of quantitation(LOQ) 定量限limit test （杂质）限度（或限量）试验limutus amebocyte lysate(LAL) 鲎试验linearity and range 线性及范围linearity scanning 线性扫描liquid chromatograph/mass spectrometer (LC/MS) 液质联用仪litmus paper 石蕊试纸loss on drying 干燥失重low pressure gradient pump 低压梯度泵luminescence 发光lyophilization 冷冻干燥Mmain constituent 主成分make-up gas 尾吹气maltol reaction 麦牙酚试验Marquis test 马奎斯试验mass analyzer detector 质量分析检测器mass spectrometric analysis 质谱分析mass spectrum 质谱图mean deviation 平均偏差measuring flask, volumetric flask 量瓶measuring pipet(te) 刻度吸量管medicinal herb 草药melting point 熔点melting range 熔距metabolite 代谢物metastable ion 亚稳离子methyl orange 甲基橙methyl red 甲基红micellar chromatography 胶束色谱法micellar electrokinetic capillary chromatography(MECC, MEKC) 胶束电动毛细管色谱法micelle 胶束microanalysis 微量分析microcrystal 微晶microdialysis 微透析micropacked column 微型填充柱microsome 微粒体microsyringe 微量注射器migration time 迁移时间millipore filtration 微孔过滤minimum fill 最低装量mobile phase 流动相modifier 改性剂，调节剂molecular formula 分子式monitor 检测，监测monochromator 单色器monographs 正文mortar 研钵moving belt interface 传送带接口multidimensional detection 多维检测multiple linear regression 多元线性回归multivariate calibration 多元校正Nnatural product 天然产物Nessler glasses(tube) 奈斯勒比色管Nessler‘s reagent 碱性碘化汞钾试液neutralization 中和nitrogen content 总氮量nonaqueous acid-base titration 非水酸碱滴定nonprescription drug, over the counter drugs (OTC drugs) 非处方药nonproprietary name, generic name 非专有名nonspecific impurity 一般杂质non-volatile matter 不挥发物normal phase 正相normalization 归一化法notice 凡例nujol mull method 石蜡糊法Ooctadecylsilane chemically bonded silica 十八烷基硅烷键合硅胶octylsilane 辛（烷）基硅烷odorless 无臭official name 法定名official specifications 法定标准official test 法定试验on-column detector 柱上检测器on-column injection 柱头进样on-line degasser 在线脱气设备on the dried basis 按干燥品计opalescence 乳浊open tubular column 开管色谱柱optical activity 光学活性optical isomerism 旋光异构optical purity 光学纯度optimization function 优化函数organic volatile impurities 有机挥发性杂质orthogonal function spectrophotometry 正交函数分光光度法orthogonal test 正交试验orthophenanthroline 邻二氮菲outlier 可疑数据，逸出值overtones 倍频峰，泛频峰oxidation-reduction titration 氧化还原滴定oxygen flask combustion 氧瓶燃烧Ppacked column 填充柱packing material 色谱柱填料palladium ion colorimetry 钯离子比色法parallel analysis 平行分析parent ion 母离子particulate matter 不溶性微粒partition coefficient 分配系数parts per million (ppm) 百万分之几pattern recognition 模式识别peak symmetry 峰不对称性peak valley 峰谷peak width at half height 半峰宽percent transmittance 透光百分率pH indicator absorbance ratio method pH指示剂吸光度比值法pharmaceutical analysis 药物分析pharmacopeia 药典pharmacy 药学phenolphthalein 酚酞photodiode array detector(DAD) 光电二极管阵列检测器photometer 光度计pipeclay triangle 泥三角pipet(te) 吸移管，精密量取planar chromatography 平板色谱法plate storage rack 薄层板贮箱polarimeter 旋光计polarimetry 旋光测定法polarity 极性polyacrylamide gel 聚丙酰胺凝胶polydextran gel 葡聚糖凝胶polystyrene gel 聚苯乙烯凝胶polystyrene film 聚苯乙烯薄膜porous polymer beads 高分子多孔小球post-column derivatization 柱后衍生化potentiometer 电位计potentiometric titration 电位滴定法precipitation form 沉淀形式precision 精密度pre-column derivatization 柱前衍生化preparation 制剂prescription drug 处方药pretreatment 预处理primary standard 基准物质principal component analysis 主成分分析programmed temperature gas chromatography 程序升温气相色谱法prototype drug 原型药物provisions for new drug approval 新药审批办法purification 纯化purity 纯度pyrogen 热原pycnometric method 比重瓶法Qquality control(QC) 质量控制quality evaluation 质量评价quality standard 质量标准quantitative determination 定量测定quantitative analysis 定量分析quasi-molecular ion 准分子离子Rracemization 消旋化radioimmunoassay 放射免疫分析法random sampling 随机抽样rational use of drug 合理用药readily carbonizable substance 易炭化物reagent sprayer 试剂喷雾器recovery 回收率reference electrode 参比电极refractive index 折光指数related substance 有关物质relative density 相对密度relative intensity 相对强度repeatability 重复性replicate determination 平行测定reproducibility 重现性residual basic hydrolysis method 剩余碱水解法residual liquid junction potential 残余液接电位residual titration 剩余滴定residue on ignition 炽灼残渣resolution 分辨率，分离度response time 响应时间retention 保留reversed phase chromatography 反相色谱法reverse osmosis 反渗透rider peak 驼峰rinse 清洗，淋洗robustness 可靠性，稳定性routine analysis 常规分析round 修约（数字）ruggedness 耐用性Ssafety 安全性Sakaguchi test 坂口试验salt bridge 盐桥salting out 盐析sample applicator 点样器sample application 点样sample on-line pretreatment 试样在线预处理sampling 取样saponification value 皂化值saturated calomel electrode(SCE) 饱和甘汞电极selectivity 选择性separatory funnel 分液漏斗shoulder peak 肩峰signal to noise ratio 信噪比significant difference 显著性差异significant figure 有效数字significant level 显著性水平significant testing 显著性检验silanophilic interaction 亲硅羟基作用silica gel 硅胶silver chloride electrode 氯化银电极similarity 相似性simultaneous equations method 解线性方程组法size exclusion chromatography(SEC) 空间排阻色谱法sodium dodecylsulfate, SDS 十二烷基硫酸钠sodium hexanesulfonate 己烷磺酸钠sodium taurocholate 牛璜胆酸钠sodium tetraphenylborate 四苯硼钠sodium thiosulphate 硫代硫酸钠solid-phase extraction 固相萃取solubility 溶解度solvent front 溶剂前沿solvophobic interaction 疏溶剂作用specific absorbance 吸收系数specification 规格specificity 专属性specific rotation 比旋度specific weight 比重spiked 加入标准的split injection 分流进样splitless injection 无分流进样spray reagent （平板色谱中的）显色剂spreader 铺板机stability 稳定性standard color solution 标准比色液standard deviation 标准差standardization 标定standard operating procedure(SOP) 标准操作规程standard substance 标准品stationary phase coating 固定相涂布starch indicator 淀粉指示剂statistical error 统计误差sterility test 无菌试验stirring bar 搅拌棒stock solution 储备液stoichiometric point 化学计量点storage 贮藏stray light 杂散光substituent 取代基substrate 底物sulfate 硫酸盐sulphated ash 硫酸盐灰分supercritical fluid chromatography(SFC) 超临界流体色谱法support 载体（担体）suspension 悬浊液swelling degree 膨胀度symmetry factor 对称因子syringe pump 注射泵systematic error 系统误差system model 系统模型system suitability 系统适用性Ttablet 片剂tailing factor 拖尾因子tailing peak 拖尾峰tailing-suppressing reagent 扫尾剂test of hypothesis 假设检验test solution(TS) 试液tetrazolium colorimetry 四氮唑比色法therapeutic drug monitoring(TDM) 治疗药物监测thermal analysis 热分析法thermal conductivity detector 热导检测器thermocouple detector 热电偶检测器thermogravimetric analysis(TGA) 热重分析法thermospray interface 热喷雾接口The United States Pharmacopoeia(USP) 美国药典The Pharmacopoeia of Japan(JP) 日本药局方thin layer chromatography(TLC) 薄层色谱法thiochrome reaction 硫色素反应three-dimensional chromatogram 三维色谱图thymol 百里酚（麝香草酚）（指示剂）thymolphthalein 百里酚酞（麝香草酚酞）（指示剂）thymolsulfonphthalein ( thymol blue) 百里酚蓝（麝香草酚蓝）（指示剂）titer, titre 滴定度time-resolved fluoroimmunoassay 时间分辨荧光免疫法titrant 滴定剂titration error 滴定误差titrimetric analysis 滴定分析法tolerance 容许限toluene distillation method 甲苯蒸馏法toluidine blue 甲苯胺蓝（指示剂）total ash 总灰分total quality control(TQC) 全面质量控制traditional drugs 传统药traditional Chinese medicine 中药transfer pipet 移液管turbidance 混浊turbidimetric assay 浊度测定法turbidimetry 比浊法turbidity 浊度Uultracentrifugation 超速离心ultrasonic mixer 超生混合器ultraviolet irradiation 紫外线照射undue toxicity 异常毒性uniform design 均匀设计uniformity of dosage units 含量均匀度uniformity of volume 装量均匀性（装量差异）uniformity of weight 重量均匀性（片重差异）Vvalidity 可靠性variance 方差versus …对…，…与…的关系曲线viscosity 粘度volatile oil determination apparatus 挥发油测定器volatilization 挥发法volumetric analysis 容量分析volumetric solution(VS) 滴定液vortex mixer 涡旋混合器Wwatch glass 表面皿wave length 波长wave number 波数weighing bottle 称量瓶weighing form 称量形式weights 砝码well-closed container 密闭容器Xxylene cyanol blue FF 二甲苯蓝FF（指示剂）xylenol orange 二甲酚橙（指示剂）Zzigzag scanning 锯齿扫描zone electrophoresis 区带电泳zwitterions 两性离子zymolysis 酶解作用。

Remaining useful life estimation - A review on the statistical data driven approaches

⇑ Corresponding authors at: Salford Business School, University of Salford, Salford
M5 4WT, UK. Tel.: +44 0161 2954124; fax: +44 0161 2954947 (W. Wang); tel.: +86 010 62794461; fax: +86 010 62786911 (D.-H. Zhou).
a Department of Automation, Xi’an Institute of Hi-Tech, Xi’an 710025, Shaanxi, China b Salford Business School, University of Salford, Salford M5 4WT, UK c Department of Automation, TNLIST, Tsinghua University, Beijing 100084, China d School of Economics and Management, Beijing University of Science and Technology, China e PHM Centre of City University of Hong Kong, Hong Kong
The RUL of an asset is clearly a random variable and it depends on the current age of the asset, the operation environment and the observed condition monitoring (CM) or health information. Deﬁne Xt as the random variable of the RUL at time t (age or usage), then the probability density function (PDF) of Xt conditional on Yt is

水溶性膦酸双核金属钌催化t-BuOOH氧化醇性能

第34 5期2018 5 月无机化学学报CHINESE JOURNAL OF INORGANIC CHEMISTRYVol.34 No.5 883-888水溶性膦酸双核金属钌催化!-BuOOH 氧化醇性能廖海深1李辰1易小艺！，&，2中南大学化学化工学院，长沙410083))2锰资源高效清洁利用湖南省重点实验室，长沙410083)摘要：双核金属钌膦酸配合物)NH4)3tR ：2)hedP)2&2H2〇)hedP=羟基亚乙基二膦酸)在常温水溶性体系中，无需相转移催化剂，可以高效催化！-BuOOH 氧化各种醇类，包括一级醇、二级醇和长链醇。

催化机理研究表明，双核金属钌膦酸配合物的GR^-Ru1』3-单元与！-BuOOH 作用，可以形成含过氧键的Rum -Rum -OO!Bu 过渡态，然后再对底物醇氧化。

该机，如发现酸抑制而碱催进该反，紫外可见光谱表明Rum -Rum -OO!B u 的形成。

关键词：双核钌；金属-金属键；醇氧化；水溶性体系中图分类号：O643; O614.82+1 文献标识码：A文章编号：1001-4861(2018)05-0883-06DOI: 10.11862/CJIC.2018.128!er!-Butyl Hydroperoxide Oxygenation of Alcohol Catalyzed byDiruthenium Diphosphonate in Aqueous SolutionLIAO Hai-Shen1 LI Chen1 YI Xiao-Yi* *12(^College of Chemistry and Chemical Engineering, Central South University, Changsha 410083, China) (^Hunan provincial Key Laboratory of E fficient and Clear Utilization of M anganese Resources,Central South University, Changsha 410083, China)Abstract : (NH4)3[Ru2(hedp)2].2H2O (hedp=1-hydroxyethylidenediphosphonate) with t-BuOOH as oxidant is provedto be highly effective in the catalytic oxidation of alcohol, such as secondary alcohol and long-chain aliphaticalcohol, in aqueous solution at room temperature. The phase-transfer catalyst is not necessary. The acid or the coordinated base inhibits ketone production, while the base improves the reaction selectivity and the yield. The mechanism is proposed as a peroxometal pathway. The UV-Vis spectra prove that the key intermediates Ru 2- OO!B u is formed.Keywords ： dinuclear ruthenium; metal-metal bond; alcohol oxidation; aqueous solution醇氧化的反，在化工、制药可忽视的地位，科学一现、氧化的，用、可用的催化剂，环境友好的O 2，H 2O 2作氧化剂，而在温和件，水溶液体系中一氧化过.是追求目标一。

Studies on dissolution testing of the nifedipine gastrointestinal therapeutic system. I. Description

Journal of Controlled Release 48(1997)1–8Studies on dissolution testing of the nifedipine gastrointestinaltherapeutic system.I.Description of a two-phase in vitrodissolution testa ,b b b *John S.Grundy ,Keith E.Anderson ,James A.Rogers ,Robert T.Fostera Elanco Animal Health ,A Division of Eli Lilly and Company ,2001West Main Street ,P .O .Box 708,Greenﬁeld ,Indiana 46140,USAb Faculty of Pharmacy and Pharmaceutical Sciences ,University of Alberta ,Edmonton ,Alberta ,Canada T 6G 2N 8Received 12August 1996;received in revised form 3January 1997;accepted 15January 1997AbstractThe nifedipine gastrointestinal therapeutic system (GITS)incorporates a push–pull osmotic pump to release–in zero-order fashion–a ﬁnely-divided suspension of nifedipine,which must then undergo dissolution in the GI tract before the drug can be absorbed.Classical,differential (ALZA)and ﬂow-through type dissolution methods adequately characterize the in vitro nifedipine suspension release rate from the nifedipine GITS:however,these methods fail to measure the in vitro dissolution rate of the suspended particles–a potentially signiﬁcant shortcoming considering that nifedipine is poorly water-soluble (#1021m g ml ).Therefore,an in vitro two-phase dissolution system was developed.This system measured the rate of nifedipine ‘transfer’from an aqueous phase (simulated intestinal ﬂuid,USP,without pancreatin containing a nifedipine GITS tablet)into an organic phase (n -octanol),a process dependent on release of the drug suspension from the tablet,dissolution in the aqueous phase and partitioning in the organic phase.For the 30mg and 60mg nifedipine GITS formulations tested the two-phase method indicated that about 90%of the drug was ‘transferred’within 30h.This is in contrast to the results from single-phase dissolution methods showing that about 90%of drug is ‘released’within 24h.Results obtained from the two-phase dissolution method appear to be in better agreement with published in vivo studies of the nifedipine GITS with regard to the rate and duration of nifedipine absorption from the GI tract.This emphasizes the importance of differentiating between drug release and drug dissolution for this type of formulation:but,the two-phase dissolution method may also be useful for other pharmaceutical formulations and poorly water-soluble drugs.©1997Elsevier Science Ireland Ltd.Keywords :Dissolution testing;Gastrointestinal therapeutic system;In vitro release proﬁle;Nifedipine;Octanol;Sink conditions1.Introductiondevelopment.[1]The ideal system for evaluating the drug release characteristics of poorly water-soluble The testing of pharmaceutical dosage forms for indrug formulations should be relatively simple and vitro drug release and dissolution characteristics isinexpensive,and should mirror the physiological very important for ensuring batch-to-batch qualityenvironment found in the human gastrointestinal control and to optimize formulations during product(GI)tract (i.e.,dissolution conditions should attempt to duplicate,as closely as possible,the composition,*Corresponding author.temperature and pH of the intestinal contents,as well 0168-3659/97/$17.00©1997Elsevier Science Ireland Ltd.All rights reservedPII S0168-3659(97)00064-32J.S.Grundy et al./Journal of Controlled Release48(1997)1–8as mimicking mixing of intestinal contents).Another All of these methods adequately characterize the in ideal characteristic of dissolution systems for testing vitro release rate of suspended nifedipine particles drug formulations is to maintain‘sink conditions’,from different strengths of nifedipine GITS;how-i.e.,the maximum concentration expected in an ever,the results do not appear to correlate satisfac-experiment should not exceed15%of the solubility torily with in vivo drug absorption rates.of the analyte in the dissolution medium used,[2]Qureshi et al.[1]state that compared to classical which can be a problem with poorly water-soluble dissolution systems a‘‘ﬂow-through system more21drugs such as nifedipine(#10m g ml).[3–5]closely resembles the physiological environment of These factors are particularly important in dissolu-the gastrointestinal tract,with continuous extraction tion testing of controlled-release drug formulations(removal)of drug from the dissolution vessel such as the nifedipine gastrointestinal therapeutic mimicking absorption into the systemic circulation.’’system(GITS),even though the mode of drug However,they fail to consider that for a poorly release from the GITS is considered to be indepen-water-soluble drug the dissolution of a particle dent of the physiological environment.[4,6]suspension,following tablet disintegration or release Maintaining‘sink conditions’throughout the test-from a non-disintegrating formulation(e.g.,the ing of poorly water-soluble drugs can be difﬁcult in nifedipine GITS),can be an important factor affect-single-phase dissolution systems.Techniques used to ing the absorption of a drug.In fact,in the original resolve this problem include:(1)using relatively pharmacokinetic study of the nifedipine GITS[7]it large volumes of dissolution medium or frequently is speculated that the longer lag time noted before replacing the medium(e.g.,the differential(ALZA)the start of the in vivo zero-order absorption phase method),[6](2)altering the pH of the dissolution versus the in vitro zero-order release phase is medium,(3)adding co-solvents to the medium(e.g.,probably due to a delay in the dissolution of drug in different alcohols,propylene glycol,glycerin,poly-the GI tract,as well as transport of the drug out of ethylene glycol,sorbitol),and(4)adding nonionic,the gut lumen,across the gut wall and to the cationic or anionic surfactants to the medium to systemic circulation.enhance analyte solubility.[2]However,these ap-This paper reports on the applicability of using a proaches frequently give non-physiologic dissolution two-phase dissolution system–consisting of simu-* environments.[1]Flow-through dissolution systems lated intestinalﬂuid,USP,without pancreatin(SIF) (e.g.,USP XXII apparatus4)can be used to over-and a n-octanol phase–to determine the release and come solubility limitations;but,the dissolution dissolution proﬁle of nifedipine from two different mediumﬂow-rates used may not be physiologically strengths of the nifedipine GITS.relevant.Also,for all these tests potentially rate-limiting dissolution of the suspended drug particlesmay be masked.Hence,in vitro results obtained 2.Materials and methodsfrom a single-phase dissolution system may notsatisfactorily reﬂect the in vivo drug release and 2.1.Materials and chemicalsdissolution characteristics of formulations of poorlywater-soluble drugs.The drug products tested were commercially avail-The nifedipine gastrointestinal therapeutic system able and obtained from a local pharmacy.The (GITS)is administered by the oral route and uses a formulations evaluated were30mg and60mg push–pull osmotic process designed to deliver a strengths of the nifedipine GITS(Adalat®XL,Bayer ﬁnely-divided nifedipine suspension–in a zero-order Canada,Etobicoke,ON,Canada).fashion–within the GI tract over a period of time Nifedipine powder was purchased from Sigma,St. long enough to permit once-daily dosing.[4,6].Louis,MO,USA,a methanolic solution of which21Dissolution tests which have typically been used for(100m g ml)was used as the reference standard this formulation include:a modiﬁed classical dissolu-for generating all the calibration curves in the study. tion test(USP XXII apparatus2),[6]aﬂow-through Nisoldipine(Bayer Canada,Etobicoke,ON,Canada)21method[1]and a differential(ALZA)method.[6]dissolved in methanol(100m g ml)was used as4J.S.Grundy et al./Journal of Controlled Release48(1997)1–82.5.High-performance liquid chromatographic NEC486-33MHz computer running Millennium method2010chromatography manager software Version1.1(Waters,Mississauga,ON,Canada).Nifedipine concentrations of samples preparedfrom methanolic solutions of the nifedipine GITStablet residuals(0.1ml samples diluted to1.0ml 3.Results and discussionwith HPLC grade water),and the30hour aqueousphase aliquots(1ml samples)were determined by an Both the30mg and60mg strengths of the established HPLC method(lower limit of quantita-nifedipine GITS contain,in addition to the labelled 21tion5ng ml).[9]Calibration curve standards were dose,a10%overage of drug which is not released prepared by adding a known amount of the drug to from the dosage form,[4,6]i.e.,the tablet formula-1.0ml of HPLC-grade water.A calibration curve(50tions contain33mg and66mg nifedipine,respec-to10,000ng of nifedipine)was determined from the tively.The total recovery of nifedipine from each of best-ﬁt regression line calculated using a1/x weight-the GITS tablets tested using the two-phase dissolu-ing factor(where x corresponds to the amount of tion system are shown in Table1,and are in 2nifedipine added;r.0.999).HPLC instrumentation excellent agreement with the labelled quantities included:a Model600E solvent delivery system,a including the10%overage(mean recovery was Model717autosampler;a Model486tunable UV–about97%).Hence,no signiﬁcant loss or degra-VIS absorbance detector(set to350nm);and an dation of nifedipine occurred during the collection,Table1aTotal Recovery of Nifedipine from each Nifedipine GITS Tablet at30hNifedipine Amount of Nifedipine Amount of Amount of TotalbGITS transferred(mg)Nifedipine Nifedipine in Recovery(mg)cformulation[%of releasable dose]remaining in the the aqueous[%ofd e ftablet(mg)phase(mg)theoretical] 30mg strengthTab125.90[88.8] 6.130.3732.40[98.2] Tab224.87[84.6]7.510.3032.68[99.0] Tab324.07[84.6]7.230.3131.61[95.8] Tab424.34[84.2]7.340.4332.11[97.3] Tab523.24[82.4]7.720.3831.34[95.0] Tab625.40[86.4] 6.920.3632.68[99.0] Mean6s524.6460.967.1460.560.3660.0532.1460.56[85.262.2][97.461.7]60mg strengthTab152.46[93.4]8.93 1.0062.39[94.5] Tab253.87[97.3] 6.880.78661.54[93.2] Tab350.45[85.5]14.590.5465.58[99.4] Tab453.46[89.2]11.78 1.3866.62[100.9] Tab553.51[93.8]8.85 1.0563.41[96.1] Tab657.43[100] 5.03 1.0263.48[96.2] Mean6s553.5362.289.3463.420.9660.2863.8461.92[93.265.3][96.762.9]a Mean data are presented as arithmetic means6standard deviation(s).b Amount of nifedipine transferred from the aqueous phase to the organic phase in30h.c Calculated from the amount of nifedipine transferred divided by90%of the total amount of nifedipine recovered(see text).d Amount of nifedipine remaining or unreleased from the nifedipine GITS tablet by30h.e Amount of nifedipine in the aqueous phase at30h.f Based on the labeled quantity of nifedipine plus10%overage as claimed by the manufacturer.J.S.Grundy et al./Journal of Controlled Release48(1997)1–87 and nimodipine.[11]In this method,aqueous disso-one-piece customized unit could be designed and lution medium is pumped through aﬂow-through developed.cell(of variable design)in which a drug becomes Likewise,using the basket method(USP XXII partially dissolved.In a separate extraction compart-apparatus1)did not provide enough mixing at the ment,a chloroform phase is used to extract the interface between the organic and aqueous phases,or dissolved drug and the two phases are subsequently prevent the nifedipine suspension released from separated.The chloroform serves as both a sink for falling to the bottom of the dissolution vessel.On the the slightly soluble drug substance and is used for other hand,the hybrid stirrer used inﬁnal method the spectrophotometric determination of the com-enabled streamlined hydrodynamicﬂow of dissolu-pound of interest.tion medium adjacent to all sides of the tablet, The choice of n-octanol–a solvent often used to adequate stirring of the two phases,and the degree of calculate the partition coefﬁcients of drugs–as the mixing obtained did not allow nifedipine particles to organic phase in the two-phase system described in settle at the bottom of the dissolution vessel.At the this report was based on its desirable physical–same time,the integrity of the planar oil/water chemical properties,including:(1)n-octanol is interface was not altered at the stirring rate used, practically insoluble in water(0.05g/100g H O);avoiding irregular mixing of the two phases.Hence,2[12](2)n-octanol is less dense than water(speciﬁc n-octanol did not come into contact with the gravity0.825at208C),[12]permitting ease of nifedipine GITS tablet at any time ensuring that the sampling;(3)n-octanol possesses low volatility partitioning process of nifedipine was not compro-(b.p.51958C),[12]hence n-octanol will not readily mised.evaporate at378C and thus a relatively constant There is a paucity of reports in the literature upper phase volume can be maintained;and(4)comparing in vitro nifedipine dissolution with in nifedipine possesses a high n-octanol:water distribu-vivo absorption and efﬁcacy for any of the nifedipine tion coefﬁcient of about10,000.[3,10]Substituting formulations currently available(e.g.,immediate-re-chloroform for n-octanol was also attempted(results lease,prolonged-action,sustained-release,GITS, not reported),however this solvent possesses less etc.).[15]Takahashi et al.[5]showed that the in desirable physical–chemical properties[i.e.,greater vivo drug absorption pattern of nifedipine from a soft water solubility(0.742g/100g H2O),and it is more gelatin capsule(5mg)is best simulated by the in dense than water(speciﬁc gravity1.484)][13]and vitro dissolution pattern obtained using the rotating ultimately gave unsatisfactory results.dialysis cell(RDC)method,which uses a cell A basket-paddle hybrid stirrer was used in this containing a buffered solution paired with n-octanol study(Fig.1),primarily because it was observed that as the dissolution medium.Alternatively,the advan-a paddle stirrer(USP XXII apparatus2)did not tages of the two-phase system proposed here for displace,to our satisfaction,the nifedipine GITS predicting in vivo drug absorption are simplicity and tablet from its resting position at the bottom of the theﬂexibility of organic and aqueous phase ratios to dissolution vessel.This caused concern that imbibi-accommodate different types of formulations and tion of water by the tablet may be inhibited(i.e.,due drugs.to a reduction in the exposed surface area of the Comparison of the nifedipine transfer rate-time tablet),or could lead to occlusion of the precision proﬁles obtained in this study(Fig.2)with the laser drilled hole through which the nifedipine plasma nifedipine concentration–time proﬁles ob-suspension is released,[4,6,14]or that some of the tained in several human clinical trials[4,7,16,17] released suspension could be entrapped beneath the appears to suggest an improved in vivo–in vitro tablet.It should be noted,however,that a separate correlation.Measurable plasma nifedipine concen-experiment using the paddle stirrer alone was not trations have been reported for up to36h in some done,thus the results of both methods could not be cases.[4,7]This suggests that drug absorption compared.On the assumption that a hybrid system of extends to at least30h using the GITS,given the the type described here is required and the method fact that nifedipine has a relatively short plasma was adopted routinely for the nifedipine GITS,a half-life in young healthy subjects(1to2h).8J.S.Grundy et al./Journal of Controlled Release48(1997)1–8[5]M.Takahashi,M.Mochizuki,T.Itoh,M.Ohta,Studies on [4,18,19]Thus,the shorter duration of drug releasedissolution tests for soft gelatin capsules.IV.Dissolution test (,30h)determined with single-phase dissolutionof nifedipine soft gelatin capsule containing water soluble methods does not correlate satisfactorily with in vivovehicles by the rotating dialysis cell method.Chem.Pharm. drug absorption proﬁles(.30h).The rate of in vivo Bull.42(2)(1994)333–336.drug absorption also appears to be considerably less[6]D.R.Swanson, B.L.Barclay,P.S.Wong, F.Theeuwes,Nifedipine gastrointestinal therapeutic system.Am.J.Med. than that predicted from the rate of in vitro drug83(Suppl.6B)(1987)3–9.parison of the data obtained with the[7]M.Chung,D.P.Reitberg,M.Gaffney,W.Singleton,Clinical two-phase dissolution method with pharmacokineticpharmacokinetics of nifedipine gastrointestinal therapeutic data obtained from a human clinical trial with both system.A controlled-release formulation of nifedipine.Am. 30mg and60mg nifedipine GITS tablets is the topic J.Med.83(Suppl.6B)(1987)10–14.[8]J.S.Grundy,R.Kherani,R.T.Foster,Photostability de-of part II of this report.termination of commercially available nifedipine oral dosage In summary,the application of a relatively simpleformulations.J.Pharm.Biomed.Anal.12(1994)1529–and inexpensive two-phase dissolution test for1535.evaluating drug release and dissolution characteris-[9]J.S.Grundy,R.Kherani,R.T.Foster,Sensitive high-per-tics of the nifedipine GITS was demonstrated by formance liquid chromatographic assay for nifedipine inhuman plasma utilizing ultraviolet detection.J.Chromatogr. slightly modifying existing dissolution apparatus.B654(1994)146–151.The modiﬁed dissolution test described here has[10]R.S.Chaudhary,S.S.Gangwal,V.K.Gupta,Y.N.Shah,K.C. potential applicability for improved prediction of inJindal,S.Khanna,Dissolution system for nifedipine sus-vivo performance of the nifedipine GITS.Also,it tained release formulations.Drug Dev.Ind.Pharm.20(7) could be adapted for use with GITS or similar(1994)1267–1274.[11]E.Porges,B.Schade,W.Ropte,Automatedﬂow-through pharmaceutical formulations of other poorly water-method to determine the dissolution rate of slightly soluble soluble drugs.substances.Pharm.Ind.47(1)(1985)77–86.[12]T.W.Graham Solomons,Alcohols,phenols,and ethers,in:Organic Chemistry,second ed.,John Wiley,Toronto,1980, Acknowledgmentsp.646.[13]S.Budavari,The Merck Index:an encyclopedia of chemi-The authors would like to thank Dr.Fred F.cals,drugs and biologicals,eleventh ed.,Merck,Rahway, Cantwell(Dept.of Chemistry,University of Alberta,NJ,1989.[14]M.Shapiro,M.A.Jarema,S.Gravina,Magnetic resonance Edmonton,AB)for his helpful suggestions duringimaging of an oral gastrointestinal-therapeutic-system the writing of this manuscript.Dr.John S.Grundy(GITS)tablet.J.Contr.Rel.38(1996)123–127.was a recipient of a Pharmacy Manufacturers As-[15]A.C.Mehta,S.Hart-Davies,E.A.Kay,In vitro dissolution sociation of Canada-Health Research Foundation/studies on nifedipine capsules.J.Clin.Pharm.Ther.20(4) Medical Research Council of Canada(PMAC-HRF/(1995)243–245.[16]R.Schneider,D.Stolero,L.Griffel,R.Kobelt,E.Brendel, MRC)Studentship.A.Iaina,Pharmacokinetic proﬁle of nifedipine GITS inhypertensive patients with chronic renal impairment.Drugs48(Suppl.1)(1994)16–22.References¨[17]P.Crome,F.O.Muller,P.Wijayawardhana,G.Groenewoud,H.K.L.Hundt,G.Leighton,H.G.Luus,R.Schall,M.van´[1]S.A.Qureshi,G.Caille,R.Brien,G.Piccirilli,V.Yu,I.J.Dyk,Single dose and steady-state pharmacokinetic proﬁlesMcGilveray,Application ofﬂow-through dissolution method of nifedipine GITS tablets in healthy elderly and young for the evaluation of oral formulations of nifedipine.Drug volunteers.Drug Investig.5(4)(1993)193–199.Dev.Ind.Pharm.20(11)(1994)1869–1882.[18]E.M.Sorkin,S.P.Clissold,R.N.Brogden,Nifedipine.A [2]A.R.Gennaro,Remington’s Pharmaceutical Sciences,XXII,review of its pharmacodynamic and pharmacokinetic prop-Mack Publishing Company,Easton,PA,1990.erties,and therapeutic efﬁcacy,in ischaemic heart disease, [3]S.L.Ali,Nifedipine,in:K.Florey(Ed.),Analytical Proﬁles hypertension and related cardiovascular disorders.[Review].of Drug Substances,Vol.18,Academic Press,New York,Drugs30(3)(1985)182–274.1989,pp.221–288.[19]C.H.Kleinbloesem,P.van Brummelen, D.D.Breimer, [4]J.S.Grundy,R.T.Foster,The nifedipine gastrointestinal Nifedipine.Relationship between pharmacokinetics andtherapeutic system(GITS):Evaluation of pharmaceutical,pharmacodynamics.[Review].Clin.Pharmacokinet.12(1) pharmacokinetic and pharmacological properties.Clin.Phar-(1987)12–29.macokinet.30(1)(1996)28–51.。

蛋白质磷酸化修饰的研究进展_姜铮

doi:10．3969／j．issn．1009－0002．2009．02．025综述蛋白质磷酸化修饰的研究进展姜铮，王芳，何湘，刘大伟，陈宣男，赵红庆，黄留玉，袁静中国人民解放军疾病预防控制研究所，北京100071［摘要］蛋白质磷酸化是最常见、最重要的一种蛋白质翻译后修饰方式，它参与和调控生物体内的许多生命活动。

通过蛋白质的磷酸化与去磷酸化，调控信号转导、基因表达、细胞周期等诸多细胞过程。

随着蛋白质组学技术的发展和应用，蛋白质磷酸化的研究越来越受到广泛的重视。

我们介绍了蛋白质磷酸化修饰的主要类型与功能、磷酸化蛋白质分析样品的富集及制备、磷酸化蛋白的鉴定及磷酸化位点的预测、蛋白分离后磷酸化蛋白的检测，及蛋白质磷酸化的分子机制，并综述了近年来国内外的主要相关研究进展。

［关键词］磷酸化修饰；磷酸化蛋白鉴定；磷酸化位点检测［中图分类号］Q52［文献标识码］A［文章编号］1009－0002(2009)02－0233－05Progress on Protein／Peptide PhosphorylationJIANG Zheng,WANG Fang,HE Xiang,LIU Da－Wei,CHEN Xuan－Nan,ZHAO Hong－Qing,HUANG Liu－Yu,YUAN JingInstitute of Disease Control and Prevention,Academy of Military Medical Sciences,100071Beijing,China［Abstract］Phosphorylation is one of the most important post－translational modifications of proteins,which is related to many activities of life．By reversible protein phosphorylation eukaryotes control many cellular processes including signal transduction,gene expression,and the cell cycle etc．As the development and application of the proteomics,the studies of the protein phosphorylation have become more important．This article has introduced the main types and functions of the protein phosphorylation,the enrichment and preparation of phosphoproteins and phosphopeptides,the identification of the phosphopeptides,the determination and prediction of the specific－phosphorylation－site,the phosphorelated modifications of the proteins,and the progress on studies above as well．［Key words］phosphorelated modifications;identification of the phosphopeptides;determination of the specific－phosphory-lation－site几乎所有的蛋白质在合成过程中或合成后都要经过某些形式的翻译后修饰，一些不合适的修饰常常与疾病相关，某些特定的翻译后修饰还被作为疾病的生物标志或治疗的靶标。

中文翻译

Proceedings of the Combustion InstituteV olume 32, Issue 1, 2009, Pages 229-237doi:10.1016/j.proci.2008.05.005 | How to Cite or Link Using DOICopyright © 2009 Elsevier Inc. All rights reserved.Permissions & ReprintsA chemical kinetic study of n-butanol oxidation at elevated pressure in a jet stirred reactorP. Dagauta, S.M. Sarathyb and M.J. Thomsonb, ,aCNRS, 1C, Avenue de la recherche scientifique, 45071 Orléans Cedex 2, France bDepartment of Mechanical and Industrial Engineering, University of Toronto, 5 King’s College Road, Toronto, Ont., Canada M5S 3G8Available online 20 September 2008.AbstractBiofuels are attractive alternatives to petroleum derived transportation fuels. n-Butanol,or biobutanol, is one alternative biofuel that can replace gasoline and diesel in transportation applications. Similar to ethanol, n-butanol can be produced via the fermentation of sugars, starches, and lignocelluloses obtained from agricultural feedstocks. n-Butanol has several advantages over ethanol, but the detailed combustion characteristics are not well understood. This paper studies the oxidation of n-butanol in a jet stirred reactor at 10 atm and a range of equivalence ratios. The profiles for CO, CO2, H2O, H2, C1–C4 hydrocarbons, and C1–C4 oxygenated compounds are presented herein. High levels of carbon monoxide, carbon dioxide, water, hydrogen, methane, formaldehyde, ethylene, and propene are detected. The experimental data are used to validate a novel detailed chemical kinetic mechanism for n-butanol oxidation. The proposed mechanism well predicts the concentration of major product species at all temperatures and equivalence ratios studied. Insights into the prediction of other species are presented herein. The proposed mechanism indicates that n-butanol consumption is dominated by H-atom abstraction from the α, β, and γ carbon atoms. A sensitivity analysis is also presented to show the effects of reaction kinetics on the concentration of several poorly predicted species. Keywords: n-Butanol; 1-Butanol; Jet stirred reactor; Kinetic modeling; Reaction mechanism Article Outline1.Introduction2.Experimental methods3.Computational methods4.Results and discussion5.ConclusionsAcknowledgementsAppendix A.Supplementary dataReferences1. IntroductionA potential biofuel for use in both gasoline and diesel engines is n-butanol.Historically, industrial scale production of n-butanol from biomass feedstocks was the second largest fermentation process, exceeded only by ethanol. However, its demise was brought about in the early 1960s when petroleum derived n-butanol became more economically feasible [1]. Recent advances in n-butanol production in the laboratory have spurred interest in commercial scale production of the n-butanol[2] and [3]. Recently, BP and Dupont announced that they would commercially produce n-butanol,which they call biobutanol, as a gasoline blending component for automotive fuels [4] and [5]. n-Butanol is produced via a fermentation process similar to that of ethanol, and therefore its feedstocks could include sugar beet, sugar cane, corn, wheat and also cellulosic biomass. n-Butanol has several advantages over ethanol including enhanced tolerance to water contamination allowing the use of existing distribution pipelines, the ability to blend at higher concentrations without retrofitting vehicles, and better fuel economy.Relatively few engine studies of n-butanol have been published. Yacoub et al. used gasoline blended with a range of C1–C5 alcohols (including n-butanol)to fuel a single-cylinder spark ignition (SI) engine [6]. They found that the n-butanol blends had less knock resistance than neat gasoline. The n-butanol blends also had reduced CO and hydrocarbon emissions but increased NOx emissions. This may be due to the n-butanol blends having a higher flame temperature and earlier spark timing. Of particular interest to the present study is that the primary oxygenated hydrocarbon emissions were n-butanol,formaldehyde and to a lesser extent, acetaldehyde. A study by Miller et al. successfully operated unmodified gasoline and diesel engines on blends containing 0–20% n-butanol in gasoline and 0–40% n-butanol in diesel fuel [7]. Another study successfully ran a compression ignition (CI) engine fueled with n-butanol and diesel fuel microemulsions [8].Predictive models provide a better understanding of the combustion performance and emissions characteristics of biofuel compositions and why they differ from petroleum derived materials. The development of an n-butanol model requires understanding of its fundamental pyrolysis and oxidation kinetics. However, few studies have examined the combustion chemistry of n-butanol, while none have developed a detailed chemical kinetic mechanism of the fuel. A 1959 study by Barnard examined the pyrolysis of n-butanol[9]. The experiments were carried out in a static reactor at temperatures between 579 and 629 °C. Barnard suggested that, in the absence of oxygen, n-butanol primarily reacts by the fission of the molecule at the C3H7–CH2OH bond. This produces formaldehyde, ethylene and a methyl radical, following the decomposition of the n-propyl radical. Barnard also conducted a similar study of t-butanol[10]. A study by Roberts measured the burning velocities of n-butanol using schlieren photographs of the flames [11], and found that the maximum burning velocity of n-butanol is similar to that of isopropyl alcohol and isopentyl alcohol. A recent study by McEnally and Pfefferle [12] measured the temperature and species in an atmospheric-pressure coflowing laminar nonpremixed flames. The fuels consisted of methane doped with one of the four isomers of butanol.They claimed that unimolecular dissociation was dominant, not H-atom abstraction. For n-butanol,this consisted of C–C fissionfollowed by β scission of the resulting radicals. In the case of n-butanol,complex fission involving four-center elimination of water was estimated to account for only 1% of n-butanol decomposition. The most important measured species included ethylene (C2H4) and propene (C3H6). More recently, Yang and co-workers [13] studied laminar premixed flames fuelled by one of four isomers of butanol(including n-butanol). Their results identify combustion intermediates in the butanol flames, but do not provide concentration profiles. The qualitative data provided lends support to the aforementioned dissociation mechanism proposed by McEnally and Pfefferle [12].In this paper, we report new experimental data obtained in a jet stirred reactor (JSR) for the oxidation of n-butanol at a pressure of 10 atm and a range of equivalence ratios (0.5–2.0) and temperatures (800–1150 K). In addition, a chemical kinetic model of n-butanol is developed using the JSR experiments as validation data. Both experimental and kinetic insights are offered below.2. Experimental methodsThe JSR experimental setup used in this study has been described earlier [14] and [15]. The JSR consists of a small sphere of 4 cm diameter (39 cm3) made of fused silica (to minimize wall catalytic reactions), equipped with four nozzles of 1 mm i.d. for the admission of the gases which achieve stirring. The reactants were diluted by high-purity nitrogen (<50 ppm O2, <1000 ppm Ar, <5 ppm H2) and mixed at the entrance of the injectors. A high degree of dilution (0.1% volume of fuel) was used, reducing temperature gradients and heat release in the JSR. High-purity oxygen (99.995% pure) was used in these experiments. All the gases were preheated before injection to minimize temperature gradients inside the reactor. A regulated heating wire of ca. 1.5 kW maintained the temperature of the reactor at the desired working temperature. The n-butanol was sonically degassed before use. A Shimadzu LC10 AD VP pump with an on-line degasser (Shimadzu DGU-20 A3) was used to deliver the fuel to an atomizer–vaporizer assembly maintained at 200 °C. Good thermal homogeneity along the vertical axis of the reactor (gradients of ca. 1 K/cm) was observed for each experiment by thermo-couple (0.1 mm Pt–Pt/Rh (10%) located inside a thin-wall silica tube) measurements. The reacting mixtures were probe sampled by means of a fused silica low pressure sonic probe. The samples were analyzed online by FT-IR and off-line after collection and storage in 1 L Pyrex bulbs. Off-line analysis was done using gas chromatographs equipped with capillary columns (DB-624 and Carboplot-P7), a TCD (thermal conductivity detector), and an FID (flame ionization detector).The experiments were performed at steady state, at a constant mean residence time of 0.7 s and a constant pressure of 10 atm. The reactants were continually flowing in the reactor while the temperature of the gases inside the JSR was increased stepwise. A good repeatability was observed in the experiments and reasonable good carbon balance of 100 ± 15% was achieved.3. Computational methodsThe kinetic modeling was performed using the PSR computer code [16] that computes species concentrations from the net rate of production of each species by chemical reactions and the difference between the input and output flow rates of the species. These rates are computed from the kinetic reaction mechanism and the rate constants of the elementary reactions calculated at the experimental temperature.The reaction mechanism used here is based on a previously proposed oxidation mechanism [17], [18] and [19] for C1–C4 chemistry. Additional reactions have been added to represent thebutanol mechanism and are listed in Table 1. The oxidation of n-butanol proceeds via unimolecular initiation and hydrogen abstraction reactions. The fuel radical species formed are consumed via unimolecular decomposition (β-scission) and biomolecular reactions. Isomerization of radical species is also included in the proposed model. Table 2 presents the structure of species produced during the oxidation of n-butanol.The rate expression for new reactions derives from tabulations for alkanes and alcohols [18] and [19]. This mechanism, including references and thermochemical data, is available as Supplementary material to this article. The rate constants for reverse reactions are computed from the corresponding forward rate constants and the appropriate equilibrium constants, calculated from thermochemistry [20] and [21].Table 1. Reactions representing the oxidation of n-butanolFull-size tableNote: X denotes a radical species (OH, H, CH3, O, HCO, HO2, CH2OH, CH3O, C2H5, C2H4, C4H7, aC3H5).View Within ArticleaC4H8OHbC4H8OHcC4H8OHdC4H8OHcC3H6OHaC3H6OHFull-size tableView Within Article4. Results and discussionMolecular species concentration profiles were measured by sonic probe sampling and GC and FT-IR analyses from the oxidation of n-butanol in a JSR: hydrogen (H2), water (H2O), carbon monoxide (CO), carbon dioxide (CO2), methane (CH4), acetylene (C2H2), ethylene (C2H4), ethane (C2H6), propene (C3H6), 1-butene (C4H8), acetaldehyde (CH3HCO), formaldehyde (CH2O), butyraldehyde (C3H7CHO), and n-butanol(C4H9OH). Figure 1 presents the experimental measurements and modeling results of n-butanol obtained at = 1.0. Theexperimental results (solid symbols) show that with increasing temperature, the n-butanol levels drop significantly between 800 and 900 K. This corresponds to a large increase in the concentrations of butyraldehyde, 1-butene, and propene, all of which are products of H abstraction pathways. The concentration of these compounds then quickly decreases as the temperature increases. Ethylene, ethane, acetaldehyde, and formaldehyde concentrations are also shown to increase between 800 and 900 K. However, as the temperature increases further, the concentrations of these species tends to diminish at a slower rate than the aforementioned species.Full-size image (63K)Fig. 1. Comparison of the experimental concentration profiles obtained from the oxidation of n-butanol in a JSR at = 1, P = 10 atm, τ = 0.7 s.View Within ArticleThe following oxygenated products were detected: butanal, ethyloxirane, propanal, 2-propenal, methyloxirane, oxirane, and acetaldehyde. The oxiranes, 2-propenal, and propanal are formed at low ppm levels, and therefore no concentration profiles are reported. Enols were not detected. A comparison with results obtained for ethanol in similar conditions and keeping the initial carbon content shows butanol oxidation produces less aldehydes overall. The maximum amount ofacetaldehyde production is reduced by ca. 70% when changing the fuel from ethanol to butanol. The model predictions (open symbols with line) for = 1.0 are also shown in Fig. 1. Reasonablygood agreement is obtained for all measured species. The major product species (i.e., CO, CO2, and H2O) are well predicted by the model. Methane, ethylene, hydrogen, and formaldehyde are also reasonably well predicted across the entire temperature range. The reactivity of n-butanol is well predicted between 800 and 950 K, but at greater temperatures the reactivity is overpredicted. Species concentrations of butyraldehyde, 1-butene, and acetaldehyde are well predicted until approximately 1000 K, above which they become underpredicted. The propene concentration is underpredicted across the entire temperature range, while ethane and acetylene concentrations are overpredicted across the entire temperature range.Figure 2 presents the experimental measurements and modeling results of n-butanol obtained at = 0.5. For the most part, the experimental results show a similar trend to that observed at= 1.0. The concentration of n-butanol is lower at = 0.5 than at = 1.0 due to the fact that agreater oxygen concentration exists in the oxygen–fuel mixture. The model better predicts the concentration of most species at = 0.5 than it does at = 1.0. 1-Butene, propene, butyraldehyde,carbon monoxide, carbon dioxide, methane, ethylene, acetaldehyde, formaldehyde, water, and hydrogen are well predicted across the entire temperature range. Similar to the case of = 1.0, thereactivity of n-butanol is overpredicted above 900 K. Again, the concentrations of acetylene and ethane are overpredicted across the entire temperature range.Full-size image (64K)Fig. 2. Comparison of the experimental concentration profiles obtained from the oxidation of n-butanol in a JSR at = 0.5, P = 10 atm, τ = 0.7 s.View Within ArticleFigure 3 presents the experimental measurements and modeling results of n-butanol obtained at = 2.0. Similar trends as those observed for other equivalence ratios are observed for theexperimental data at = 2.0. At = 2.0, the reactivity of n-butanol is well predicted across theentire temperature range, something which was not observed at other equivalence ratios In addition, there is good prediction of carbon monoxide, carbon dioxide, methane, ethylene, acetaldehyde, ethane, formaldehyde, water, and hydrogen. Qualitatively, the prediction of acetylene concentration is satisfactory. The butyraldehyde concentration is well predicted below 1000 K, while above 1000 K the model underpredicts the experimental data. The concentration of 1-butene is overpredicted above 900 K, while the concentration of propene is under underpredicted across the entire temperature range.Full-size image (63K)Fig. 3. Comparison of the experimental concentration profiles obtained from the oxidation of n-butanol in a JSR at = 2, P = 10 atm, τ = 0.7 s.View Within ArticleSome general trends are observed via analysis of the data across the three equivalence ratios. The model’s prediction of carbon monoxide, carbon dioxide, methane, ethylene, for maldehyde, water, and hydrogen concentrations is reasonably accurate across all equivalence ratios. The prediction of n-butanol,acetaldehyde, and acetylene concentrations tends to improve with increasing equivalence ratio. On the other hand, an increase in equivalence ratios results in poorer prediction of 1-butene, propene, butyraldehyde, and ethane concentrations.A reaction pathway analysis was performed at = 1.0 at T = 1000 K to determine the mostdominant pathways for n-butanol consumption. Figure 4 presents the results of the analysis in diagram format, wherein heavier weight arrows represent more dominant reaction pathways. According to the proposed model, n-butanol is consumed primarily via H-atom abstraction from the α, β, and γ carbon atoms, with each pathway accounting for approximately 22% of the total n-butanol consumption. The next most dominant pathway is H-atom abstraction from the hydroxyl group, which accounts for nearly 20% of n-butanol consumption. H-atom from the δ carbon atom accounts for nearly 14% while all the unimolecular decomposition pathways combined account for less than 0.5% of n-butanol consumption. Similarly, a reaction pathway analysis at T = 1200 K showed that unimolecular decomposition accounted for less than 4% of n-butanol consumption. Therefore, it is reasonable to conclude that n-butanol consumption in the JSR is dominated by H-atom abstraction.Full-size image (16K)Fig. 4. Reaction pathway diagram for n-butanol oxidation in the JSR at = 1, P = 10 atm,τ = 0.7 s, T = 1000 K.View Within ArticleThe pathways diagram in Fig. 4 indicates that the aC4H8OH radical primarily undergoes β-scission to form acetaldehyde and an ethyl radical (C2H5). The consumption of the bC4H8OH radical is also consumed primarily by β-scission to form a hydroxyl radical and 1-butene. The cC4H8OH radical primarily undergoes β-scission to form propene and a hydroxymethyl radical(CH2OH). The hydroxymethyl radical, which is also an intermediate in several n-butanol unimolecular decomposition pathways, undergoes β-scission to create formaldehyde. The C4H9O radical, which is formed primarily via H-atom abstraction from the n-butanol hydroxyl group, undergoes β-scission to form butyraldehyde. The least prominent n-butanol H-atom abstraction pathway leads to the formation of the dC4H8OH radical, which isomerizes to form the aC4H8OH radical. The n-butanol unimolecular dissociation reactions proceed to form radical species, which then undergo β-scission to form stable species such as acetylene, ethylene, and formaldehyde, and a number of radical species.Sensitivity analyses were conducted for n-butanol,propene, and acetylene as these compounds were not always well predicted by the model. n-Butanol was underpredicted above 900 K at both = 1.0 and = 0.5. Figure 5a displays the normalized sensitivity coefficients for the top 12reactions to which the n-butanol concentration is sensitive at T = 1050 K and all equivalence ratios. A positive sensitivity coefficient implies that an incre ase in the reaction’s forward rate will increase the n-butanol concentration at the specified temperature and equivalence ratio. At all equivalence ratios, the n-butanol concentration is very sensitive to the reaction producing OH radicals via the oxidation of H radicals. At = 0.5, the n-butanol concentration is mainlysensitive to elementary reactions between hydrogen and oxygen containing species. However, at = 1.0 and = 2.0, the n-butanol concentration is more sensitive to reactions involvinghydrocarbon radical species. This is because the pool of hydrocarbon radicals becomes more predominant as the fuel concentration in the oxygen–fuel mixture increases. Of all the n-butanol consumption reactions, the n-butanol concentration is most sensitive to those involving H-abstraction by OH radicals from the α and γ carbons.Full-size image (44K)Fig. 5. Sensitivity of n-butanol and propene to select reactions in the JSR at P = 10 atm, τ = 0.7 s.View Within ArticlePropene concentrations were not well predicted at = 1.0 and = 2.0. Figure 5b displays thenormalized sensitivity coefficients for the top 11 reactions to which the propene concentration is sensitive at T = 1000 K and all equivalence ratios. The propene concentration is sensitive to elementary reactions between hydrogen and oxygen containing species, as well as reactions involving small molecular weight hydrocarbon species. In addition, the propene concentration is sensitive to n-butanol consumption reactions involving H-abstraction from the α, β, and γ carbons.A sensitivity analysis on acetylene (not in figure) indicated the acetylene concentration is sensitive to reactions involving the C2H3 radical, and to elementary reactions between hydrogen and oxygen containing species. Adjusting the reaction rates of n-butanol consumption reactions hadlittle effect on the concentration of acetylene.5. ConclusionsNew experimental data for n-butanol oxidation in a JSR at 10 atm and equivalence ratios between 0.5 and 2.0 are compared to a novel chemical kinetic model for n-butanol oxidation. The most abundant measured product species were carbon monoxide, carbon dioxide, water, hydrogen, methane, formaldehyde, ethylene, and propene. Measured in lesser amounts were butyraldehyde, 1-butene, acetaldehyde, ethane, and acetylene. The model proposed herein provides good overall agreement with the experimental data obtained across various temperatures and equivalence ratios. It is shown that H-abstraction is the major pathway of n-butanol consumption in the JSR, while unimolecular decomposition is relatively insignificant. Further model validations are still needed; they are awaiting the availability of ongoing flame measurements.AcknowledgmentThis research acknowledges funding from NSERC.References[1] D.T. Jones and D.R. Woods, Microbiol. Rev. 50 (4) (1986), pp. 484–524. View Record in Scopus | Cited By in Scopus (326)[2] T.C. Ezeji, N. Qureshi and H.P. Blaschek, Curr. Opin. Biotechnol. 18 (2007), pp. 220–227.Article | PDF (327 K) | View Record in Scopus | Cited By in Scopus (79)[3] D. Ramey, S. Yang, Production of Butyric Acid and Butanol from Biomass, Report No. DE-F-G02-00ER86106. US Department of Energy, Morgantown, Washington, 2004.[4] G. Hess, Chem. Eng. News 84 (26) (2006), p. 9. Full Text via CrossRef | View Record in Scopus | Cited By in Scopus (7)[5] G. Hess, Chem. Eng. News 85 (27) (2007), p. 8. Full Text via CrossRef | View Record in Scopus | Cited By in Scopus (1)[6] Y. Yacoub, R. Bata and M. Gautam, Proc. Inst. Mech. Eng. 212 (1998), pp. 363–379. Full Text via CrossRef | View Record in Scopus | Cited By in Scopus (15)[7] G.L. Miller, J.L. Smith and J.P. Workman, Trans. ASAE 24 (1981), pp. 538–540.[8] T.K. Bhattacharya, S. Chatterjee and T.N. Mishra, Appl. Eng. Agric. 20 (3) (2004), pp. 253–257. View Record in Scopus | Cited By in Scopus (10)[9] J.A. Barnard, Trans. Faraday Soc. 53 (11) (1957), pp. 1423–1430. Full Text via CrossRef | View Record in Scopus | Cited By in Scopus (11)[10] J.A. Barnard, Trans. Faraday Soc. 55 (6) (1959), pp. 947–951. Full Text via CrossRef | View Record in Scopus | Cited By in Scopus (4)[11] A.F. Roberts, J. Imp. Coll. Chem. Eng. Soc. 12 (1959), pp. 58–73.[12] C.S. McEnally and L.D. Pfefferle, Proc. Combust. Inst. 30 (2005), pp. 1363–1370. Article |PDF (389 K) | View Record in Scopus | Cited By in Scopus (41)[13] B. Yang, P. Oswald and Y. Li et al., Combust. Flame 148 (2007), pp. 198–209. Article | PDF (1138 K) | View Record in Scopus | Cited By in Scopus (54)[14] P. Dagaut and M. Cathonnet, Prog. Energy Combust. Sci. 32 (2006), pp. 48–92. Article |PDF (1414 K) | View Record in Scopus | Cited By in Scopus (77)[15] P. Dagaut and S. Gail, J. Phys. Chem. A 111 (19) (2007), pp. 3992–4000. Full Text via CrossRef | View Record in Scopus | Cited By in Scopus (27)[16] P. Glarborg, R.J. Kee, J.F. Grcar, J.A. Miller, PSR: A FORTRAN Program for Modeling Well-Stirred Reactors, Report No. SAND86-8209, Sandia National Laboratories, Livermore, CA, 1986.[17] T. Le Cong, P. Dagaut, Kinetics of natural gas, natural gas/syngas mixtures oxidation and effect of burnt gas recirculation: experimental and detailed modeling, in: Proceedings of the ASME Turbo Expo 2007: Power for Land, Sea and Air, Montréal, Canada, May 14–17, 2007, GT2007-27146, pp. 1–9.[18] P. Dagaut, Phys. Chem. Chem. Phys. 4 (2002), pp. 2079–2094. Full Text via CrossRef | View Record in Scopus | Cited By in Scopus (100)[19] F. Battin-Leclerc, R. Bounaceur, G.M. Côme, et al., EXGAS-ALKANES, a software for the automatic generation of mechanisms for the oxidation of alkanes, 2004, CNRS-DCPR.[20] Y. Tan, P. Dagaut, M. Cathonnet and J.C. Boettner, Combust. Sci. Technol. 102 (1994), pp. 21–55. Full Text via CrossRef[21] C. Muller, V. Michel, G. Scacchi and G.M. Côme, J. Chim. Phys. Phys.-Chim. Biol. 92 (5) (1995), pp. 1154–1178.Appendix A. Supplementary dataDownload this File (85 K)Supplementary data. The proposed n-butanol chemical kinetic mechanism in CHEMKIN format View Within ArticleDownload this File (160 K)Supplementary data. The proposed n-butanol thermochemical data in CHEMKIN formatView Within ArticleDownload this File (515 K)Supplementary data. An MS Excel data file with all the experimental and model dataView Within ArticleCorresponding author. Fax: +1 416 978 7753.Proceedings of the Combustion InstituteV olume 32, Issue 1, 2009, Pages 229-237。

药剂词汇英汉对照

药剂词汇英汉对照AAbsolute bioavailability 绝对生物利用度Absorption 吸收Acacia 阿拉伯胶Accelerated stability 加速稳定性实验Accumulation factor蓄积因子Additive 附加剂Adjuvant 辅料Administration 给药，用法Aerosil 微粉硅胶Aerosol 气雾剂Agar 琼胶Aggregation 聚集Albumin 白蛋白Alginate(alginate acid)海藻酸盐（海藻酸）Amorphous form 无定型Amphiphilic 两亲性的Angle of repose 休止角Angle of friction 摩擦角Ampoule 安瓿Anion exchange resin 阴离子交换树脂Antiseptics 防腐剂；消毒剂Antioxidant 抗氧化剂Apparent first-order absorption(elimination)表观一级吸收（消除）Apparent volume of distribution 表观分布容积Rate constant速度，速率Aromatic waters 芳香水剂Aseptic manipulation 无菌操作Autoclave 高压灭菌器Autoxidation 自动氧化Azone 氮酮BBentonite 硅皂土Binder 粘合剂Bioadhesive tablets 生物粘附片Biocompatibility 生物相容性Bioequivalence 生物等效性Biologic half-life 生物半衰期Biotransformation 生物转化Buccal tablets 口含片Buccal administration 颊给药Bulk density 堆密度CCapacity—limited and nonlinear process 能力有限和非线性过程Capillary action 毛细管作用Capping 顶裂Carboxypolymethylene 聚羧乙烯Capsul 胶囊Carnauba wax 巴西棕榈蜡Carbopol 卡波普Carrier-mediated transfer system 载体转运系统Cation exchange resin 阳离子交换树脂Cellulose acetate phthalate(CAP) 邻苯二甲酸醋酸纤维素Cetyl alcohol 鲸蜡醇Chelating agent 螯合剂Chewable tablets 咀嚼片Chipping 裂片Chitin 壳聚糖Chitosan 脱乙酰壳聚糖Chronopharmacokinetics 时辰药物动力学Clarity(test) 澄明度（检查）Cloud point 浊点Co-solvency 潜溶Coagulation 聚沉Coating 包衣Cocoa butter 可可豆脂Cohesiveness 粘着性Colloid mill 胶体磨Comminution 粉碎Comples coacervation method 复凝聚法Content uniformity test 含量均匀试验Controlled release preparation 控释制剂Coprecipitate 共沉淀物Coulter counter 库氏计数器Cracking 松片Creaming 乳析Creatinine clearance 肌酐清除率Critical micelle concentration(cmc) 临界胶束浓度Croscarmellose sodium(ccna)交联羧甲基纤维素钠Crospovidone 交联聚维酮Cyclodextrin inclusion compound 环糊精包合物DDecoction 煎煮法,汤剂Deflocculating agent 反絮凝剂Delayed release 延缓释放Dextrin 糊精Die 模圈，冲磨Differential scanning calorimetry(DSC) 差示扫描量热法Differential thermal analysis(DTA) 差示热分析法Diffusion coefficient 扩散系数Diluents 稀释剂Dimethyl acetamide(DMA) 二甲基乙酰胺Dimethyl formamide(DMF) 二甲基乙酰胺Dimethyl sulfoxide(DMSO) 二甲基亚砜Disinfectant 消毒剂，灭菌剂Disintegrant 崩解剂Dispersible tablets 分散片Displacement value 置换价Dissolution 溶出Distribution 分布Dosage form 剂型Dosage regimen 给药方案Dosing interval 给药间隔Drug compatibility 药物配伍Drug delivery system 药物传递系统Drug disposition 药物处置Duration 作用持续性Dustibility 松散性EEffervescent tablets 泡腾片Elimination 消除Elixirs酏剂Emulsions 乳剂Emulsification 乳化作用Encapsulation coefficency 包封率Endocytosis内吞作用Enemas 灌肠剂Enteric coated tablets 肠衣片Enterohepatic cycle肝肠循环Equivalent diameter 等价径，当量直径Drodible matrix 溶蚀型骨架Ethylene-vinyl acetate copolymer（EVAC）乙烯-醋酸乙烯共聚物Ethyl cellulose（EC）乙基纤维素Eudragit 丙烯酸树脂Eutectic mixture 低共熔混合物Excipient 赋形剂，辅料Extended release持续释放Extract preparation 浸出制剂Extracts 浸膏剂Extravascular administration 血管外给药FFacilitated diffusion 促进扩散Fast 禁食Feed shoe 饲粉靴Fillers 填充剂Film coated tablets 薄膜衣片Filter candle(disk) 滤棒First pass effect 首过效应Flavoring agent 矫味剂Flocculant 絮凝剂Fluctuation 波动性Fluid bed coating(granulation) 流化床包衣制粒Fluid energy mill 流能磨Fluid extracts 流浸膏剂Fluid mosaic model 液体相嵌模型Food and drug adiministration(FDA)食品药品管理局Fomulation 处方Friability 脆碎度Fusion 融合作用GGargles 漱口剂Gelatin 明胶Gelatin glycerin 甘油明胶Gildants 助流剂Glomerular filtration 肾小球滤过Glycerite 甘油剂Glyceryl monostearate 单硬脂酸甘油酯Glycoprotein 蛋白糖Good manufacturing practice(GMP)药品生产质量管理规范Granule density 粒密度Greasing bases 油脂性基质Guest molecule 客分子HHealthy volunteer 健康受试者High-efficiency particle air filter 高效空气过滤嚣Horizontal laminar flow 水平层流Hopper 加料斗Host molecule主分子Hydrocarbon base烃类基质Hydrogel matrix水凝胶骨架Hydrogenated vegetable oil氢化植物油Hydrophile-lipophile balance(HLB)亲水亲油平衡值Hydrophilic metrix亲水性骨架Hydroscopicity吸湿性Hydrotropic agent助溶剂Hydroxypropylmethyl cellulose(HPMC)羟丙基甲基纤维素Hydroxypropylstarch羟丙基淀粉IImplant植入剂In vitro体外In vivo体内Inclusion compound包合物Imcompatibility配合物Imdustrial pharmacy工业药剂学Infusion浸渍，浸剂，输注Inhalation aerosols吸入气雾剂Injection注射剂Instantraneous rate瞬时速率Interfacial polymerization界面缩聚法Intra uterine device(IUD)宫内给药嚣Intra vaginal ring(IVR)阴道环Intravenous infusion静脉滴注Inulin菊粉Iontophoresis离子等渗Irrigations灌洗剂Iso-osmotic solution等渗溶液Isotonic solution等张溶液Isotonicity等张性JThe Parmacopoeia of japan日本药局方KKidney function肾功能Kinetics parameter动力学参数LL-hydroxypropyl cellulose(L-HPC)低取代羟丙基纤维素Lactose乳糖Lag time时滞Lake色淀Laminar flow clean bench层流洁净工作台Lamination层裂Leak test(testing)漏气试验Lecithin(e)卵磷脂Linuis test鲎试验Liniment涂抹油，擦剂Liposome脂质体Loading dose负荷剂量Lotion洗剂Lubricants润滑剂Lyophilic亲液的Lyophilization冷冻干燥Lyophobic疏液的MMaceration浸渍Macromolecular solution高分子溶液Macrovesicle liposome大多孔脂质体Magnetic microspheres磁性微球Maintenance dose维持剂量Matrix tablets骨架片Mean residence time(MRT)平均滞留时间Medicinal liquor洒剂Membrane evaporator薄膜浓缩器Metastable亚稳的Methyl cellulose(MC)甲基纤维素Micelle胶团Michaelis-menten kinetics MM动力学Microcapsules微囊Microspheres 微球Microcrystalline cellulose微晶纤维素Microemulsion微乳Microencapsulation微型包囊技术Micronise微粉化Microporous membrane微孔膜Microvilli微绒毛Migration迁移Minimun effective concentration(level)最小有效浓度Minimun toxic concentration最小中毒浓度Mixer混合机Mixtures合剂Moistening agent润湿剂Monoclonal antibody单克隆抗体Mottling色斑Mucilages胶浆剂Multilamellar,multiple compartment liposome多室脂质体Multiple compartment model多室脂质体Multiple dosing多剂量给药Multiple emulsion复合型乳剂NNanocapsule毫微胶囊Nanoparticles毫微粒Nanospheres毫微球Naris drops滴鼻剂Nasal administration鼻腔给药Nonlinear pharmacokinetics非线性药物动力学National formulary美国药局方Nipagin(parabens)尼泊金OOintment软膏剂Oleaginous base油性基质One compartment model单室模型Onset起效时间Ophthalmic preparation眼用制剂Optimization technique优化技术Oral administration口服给药Oral osmotic pump口服渗透泵PParaffin石蜡Partition coefficient分配系数Passive diffusion被动扩散Passive target preparation被动靶向制剂Pastes 糊剂Peeling剥落Pellet小丸Penetration enhancer穿透促进剂Percolation渗漉Percutaneous administration经皮给药Permeability coefficient穿透系数Perogol O平平加OPH-partition theory PH-分配学说Pharmaceutics药剂学Pharmacokinetics药物动力学Pharmacopoeia药典Pharmacy药学Phase separation-coacervation相分享凝聚法Phase transition temperature相变温度Picking粘连Pill丸剂Plaster硬膏剂Plastic flow塑性流动plasticizer增塑剂Plateau concentration(level)坪浓度Pluronic F-68普朗尼克F-68Polishing,smoothing打光Poloxamer泊洛沙姆Polyacrylamide聚丙烯酰胺Polyacrylic acid聚丙烯酸Polyethylene glycols(PEC)聚乳酸Polymerization聚合Polymorphism多晶型Polymethyl methacrylate聚甲基丙烯酸甲酯Polyvinyl alcohol(PVA)聚乙烯醇Polyvinylpyrrolidone(PVP)(povidone)聚维酮Porosity孔隙率Powders散剂Preformulation处方前工作Preagelatinized starch预胶化淀粉Prescription处方Preservative防腐的Prodrug前体药物Propellants抛射剂Pseudo plastic flow伪一级反应Pulsed and regulated drug deliver system伪塑性流动Pulverization脉冲式和自调式释药系统Punch粉碎Pyrogen冲头RRate method 冲率法Rate-limiting step限速步骤Rectal dosage form直肠给药剂型Relative bioavailability相对生物利用度Residuals method殘数法Rheology流动学SSedimentation沉降Semipermeable membrane半透膜Shellac虫胶Sieve筛（子），筛（分）Sigma-minus method 总量减去法Silicone硅酮Simple coacervation method单凝聚法Single-punch press单冲压片机Sintered glass filter垂熔玻璃滤品Sodium carboxyl methyl cellulose (CMC-Na)羧甲基纤维素钠Sodium carboxyl methyl starch(CMS-Na)羧甲基淀粉纳Sodium lauryl sulfate十二烷基硫酸钠Solid dispersion固体分散体Solidification固化Solubilization增溶作用Solvate溶剂化物Sonication dispersion method超声分散法Spans司盘类Specific acid-base catalysis特殊酸碱催化Specification规格，说明书Spermacete wax鲸蜡Spirits醑剂Spray喷雾剂Stability稳定性Starch淀粉Statistical moment统计矩Steady-state稳态Stearyl alcohol十八醇Sterilization灭菌Sterilizer消毒器Sticking粘冲Sublingual tablet舌下片Sugar coated tablet糖衣片Superdisintegrants超级崩解剂Suppositories栓剂Surfactant表面活性剂Suspending agent助悬剂Suspensions混悬剂Sustained release preparation缓释制剂Sweeting agent甜味剂Syrups 糖浆剂Systemic clearance总清除率TTablets片剂Talc滑石粉Targeted drug delivery system(TDDS)靶向给药系统Terylene涤纶薄膜The area under the plasma concentration-time curve血浓时间曲线下面积The extra pharmacopoeia药学大全The international pharmacopoeia国际药典The membrane of epithelial cell上皮细胞膜The rate of gastric emptying胃排空速率Theobrome oil可可豆油Therapeutic concentration range治疗浓度范围Therapeutic equivalence治疗等效性Thixotropy触变性Time course经时过程Tincture酊剂Tragacanth西黄芪胶Transdermal therapeutic system(TTS)透皮治疗系统Transport转运Trapezoidal rule 梯形法则True density真密度Tubular reabsorption肾小管重吸收Tubular secretion肾小管分泌Tweens吐温类UUntrafiltration超滤Unilamellar,single compartment liposome单室脂质体Urinary excretion尿排泄The pharmacopeia of united states(usp)美国药典VVaseline(e)凡士林Vertical laminar flow垂直层流WWater for injection注射用水Wettability可湿性Wetting agent润湿剂ZZein 玉米朊--。

单硬脂酸甘油酯对邻苯二甲酸酯类暴露致雄性大鼠肝、肾功能损伤的影响

中图分类号:R155文献标识码：A文章编号:1002-3127(2020)06-0481-05•实验研究•单硬脂酸甘油酯对邻苯二甲酸酯类暴露致雄性大鼠肝、肾功能损伤的影响戴绮梦，杨柳，李安琪，孙悦，史弘毅，金思伊，夏玲姿，高海涛(温州医科大学公共卫生与管理学院预防医学系，浙江温州325035)【摘要】目的研究单硬脂酸甘油酯(GMS)对邻苯二甲酸酯类混合物(MIXPs)暴露致雄性大鼠肝、肾功能损伤的影响。

方法30只雄性大鼠随机分对照组.MIXPs组和MIXPs+GMS组；MIXPs组大鼠经口给予160mg/kg■bw■dMIXPs,MIXPs+GMS组大鼠经口给予160mg/kg-bw-d MIXPs和20mg/kg-bwd GMS O连续干预5周。

解剖，取肝、肾等组织，称重后取适量脏器组织，苏木素-伊红(HE)染色。

试剂盒测定血清丙氨酸转氨酶(ALT)、天冬氨酸转氨酶(AST)、尿素氮(BUN)、肌Bf(Scr)和尿酸(UA)等。

结果MIXPs组和MIXPs+GMS组大鼠体质量、肝和肾质量依次降低，肝、肾组织结构损伤依次加剧，主要表现为MIXPs组肝组织有轻微脂肪变、肾小管上皮细胞凋亡、坏死,MIXPs+GMS组肝组织贮脂细胞增生，肾小管空化等；MIXPs+GMS组AST.BUN和UA明显升高。

结论GMS加剧了MIXPs对雄性大鼠肝、肾功能的损伤。

【关键词】邻苯二甲酸酯类；单硬脂酸甘油酯；乳化剂；肝功能；肾功能Glycerin monostearate exacerbates phthalates5toxicity to liver function andrenal function in male ratsDAI Qi-meng,YANG Liu,LI An-qi,SUN Yue,SHI Hong-yi,JIN Si-yi,XIA Ling-zi,GAO Hai-tao (Department of Preventive Medicine,School of Public Health and Management,Wenzhou Medical University,Wenzhou Zhejiang325035,China)[Abstract]Objective To explore the effects of food emulsifier Glycerin monostearate(GMS)on the mixture of six priority controlled phthalates(MIXPs)toxicity to liver function and renal function in male rats.Methods Thirty male rats were divided randomly into three groups,normal control(NC)group,MIXPs group,MIXPs+GMS group,with10rats in each group.Rats in NC group were treated normally,while rats in MIXPs group and MIXPs+GMS group were treated with160mg/kg•bw•d MIXPs with or without20mg/kg*bwd GMS. The intervention lasted five weeks,then dissecting the rats,collecting the liver,kidney and blood.Taking appropriate amount of liver tissue and kidney tissue for pathological examination(HE staining).Serum AST,ALT,BUN,Scr and UA were measured with commercial kits. Results Compared with control,the weight of body,liver and kidney successively decreased in MIXPs group and MIXPs+GMS group,with significant decrease in MIXPs+GMS group(P<0.05or P<0.01).However,serum AST,BUN,UA increased significantly(P<0.05or P< 0.01).Pathological examination showed that hepatic steatosis,necrosis and apoptosis of renal tubular epithelial cell were found in MIXPs group,while fat-storing cells proliferation and renal tubular cavitation were found in MIXPs+GMS group.Conclusion GMS exacerbated MIXPs'toxicity to liver function and renal function in male rats.[Key words]Phthalates；Glycerin monostearate；Food emulsifier；Liver function；Renal function邻苯二甲酸酯类(phthalates,PEs)是公认的环境内分泌干扰物，可经膳食、呼吸和皮肤等途径进入人体；我国人群PEs的总暴露水平为23~159p.g/kg-bwd'"。

POLYETHER-POLYESTER-TYPE POLYPHENOL AND ITS PREPAR

专利名称：POLYETHER-POLYESTER-TYPEPOLYPHENOL AND ITS PREPARATION 发明人：NAKATSUKA RIYUUZOU,TSUGE MORIO 申请号：JP20761282申请日：19821129公开号：JPS5998128A公开日：19840606专利内容由知识产权出版社提供摘要：PURPOSE:To prepare a novel polyphenol having excellent compatibility with resins, etc., and applicable to a wide fields such as raw material of thermosetting resin, etc., by reacting an aliphatic polyol with p-hydroxybenzoic acid in a solvent successively raising the temperature of the system while removing produced water therefrom. CONSTITUTION:The objective polyphenol [e.g. a mixture of the compounds of formula (p and q are 0-5)] is prepared by reacting (A) an aliphatic polyol having >=3 valencies (e.g. glycerol, ditrimethylol propane, etc.) with (B) p-hydroxybenzoic acid using a higher ketone (e.g. cyclohexanone, acetophenone, etc.) as a solvent, raising the temperature of the system successively (preferably to the final temperature of <=200 deg.C) while removing produced water from the system. The polyphenol is composed of said polyol residues and benzoic acid residues bonded with each other through ether bonds and ester bonds, and has a number- average molecular weight of >=350 and an average ratio of the phenolic OH group to molecule of >=2.5.申请人：SUMITOMO BAKELITE KK更多信息请下载全文后查看。

PHOSPHATEBUFFER,pH72（7380）：磷酸盐缓冲液，pH值72..

PHOSPHATE BUFFER, pH 7.2 (7380)Intended UsePhosphate Buffer, pH 7.2 is used for the preparation of microbiological dilution blanks.Product Summary and ExplanationThe formula for phosphate buffer is specified by the American Public Health Association (APHA) for use in diluting test samples. Phosphate Buffer, pH 7.2 is specified for use in diluting water, dairy products and food for microbiological methods. In the examination of water1and dairy products2 the addition of magnesium chloride is recommended.This buffer is also referred to as Butterfield’s Buffered Phosphate Diluent and recommended for examination of food.2 Phosphate Buffer, pH 7.2 stabilizes the pH of water used for dilutions.Principles of the ProcedurePhosphate Buffer, pH 7.2 is used in the preparation of dilution blanks for use in microbiological testing. Phosphate Buffer is used rather than unbuffered water in order to standardize this potential variable due to the wide variation in the pH of distilled water from multiple sources.Formula / Liter SupplementPotassium Dihydrogen Phosphate ..................................... 26.22 g Magnesium Chloride, 5 mL Sodium Carbonate ............................................................... 7.78 g*Final pH: 7.2 ± 0.2 at 25°CFormula may be adjusted and/or supplemented as required to meet performance specifications.Precautions1. For Laboratory Use.2. IRRITANT. Irritating to eyes, respiratory system, and skin.DirectionsStock Solution1. Dissolve 34 g of the medium in one liter of purified water.2. Mix thoroughly.3. Autoclave at 121°C for 15 minutes. Store under refrigeration.4. *Final pH is taken after Phosphate Buffer, pH 7.2 has been autoclaved and cooled to 25°C (roomtemperature).Working Solution1. Add 1.25 mL of Stock Solution and 5 mL of a Magnesium Chloride solution (81.1 g MgCl2 6H2O per literof purified water) to purified water and make up to one liter.2. Dispense into bottles or tubes to provide 99 ± 2.0 mL, 9 ± 0.2 mL or other appropriate quantities.3. Autoclave at 121°C for 15 minutes.4. *Final pH is taken after Phosphate Buffer, pH 7.2 has been autoclaved and cooled to 25°C (roomtemperature).Quality Control SpecificationsDehydrated Appearance: Powder is homogeneous, free flowing, and white to off-white.Prepared Appearance: Prepared buffer is clear to slightly hazy / opalescent with or without trace to light precipitate and colorless.Test ProcedureRefer to appropriate references for a complete discussion and use of Phosphate Buffer, pH 7.2.ResultsRefer to appropriate references for results following test procedures.StorageStore sealed bottle containing the dehydrated medium at 2 - 30 C. Once opened and recapped, place container in a low humidity environment at the same storage temperature. Protect from moisture and light by keeping container tightly closed.ExpirationRefer to expiration date stamped on the container. The dehydrated medium should be discarded if not free flowing, or if appearance has changed from the original color. Expiry applies to medium in its intact container when stored as directed.PackagingPhosphate Buffer, pH 7.2 Code No. 7380A 500 g7380B 2 kg7380C 10 kgReferences1. Greenberg, Trussell, and Clesceri (eds.). 1985. Standard methods for the examination of water and wastewater, 16th ed.American Public Health Association, Washington, D.C.2. Richardson. (ed.). 1985. Standard methods for the examination of dairy products, 15th ed. American Public Health Association,Washington, D.C.3. Bacteriological Analytical Manual. 1995. 8th ed. AOAC International, Gaithersburg, MD.Technical InformationContact Acumedia Manufacturers, Inc. for Technical Service or questions involving dehydrated culture media preparation or performance at (517)372-9200 or fax us at (517)372-2006.。

1、下载文档前请自行甄别文档内容的完整性，平台不提供额外的编辑、内容补充、找答案等附加服务。
2、"仅部分预览"的文档,不可在线预览部分如存在完整性等问题,可反馈申请退款(可完整预览的文档不适用该条件!)。
3、如文档侵犯您的权益，请联系客服反馈,我们会尽快为您处理(人工客服工作时间：9:00-18:30)。

Phosphoenolpyruvate Availability and the Biosynthesis of Shikimic AcidSunil S.Chandran,†Jian Yi,†K.M.Draths,†Ralph von Daeniken,‡Wolfgang Weber,‡and J.W.Frost*,†Department of Chemistry,Michigan State University,East Lansing,Michigan48824-1322,andF.Hoffmann-La Roche,Ltd.,Pharmaceutical Research,CH-4070Basel,SwitzerlandThe impact of increased availability of phosphoenolpyruvate during shikimic acidbiosynthesis has been examined in Escherichia coli K-12constructs carrying plasmid-localized aroF FBR and tktA inserts encoding,respectively,feedback-insensitive3-deoxy-D-arabino-heptulosonic acid7-phosphate synthase and transketolase.Strategies forincreasing the availability of phosphoenolpyruvate were based on amplified expressionof E.coli ppsA-encoded phosphoenolpyruvate synthase or heterologous expression ofthe Zymomonas mobilis glf-encoded glucose facilitator.The highest titers and yieldsof shikimic acid biosynthesized from glucose in1L fermentor runs were achieved usingE.coli SP1.l pts/pSC6.090B,which expressed both Z.mobilis glf-encoded glucosefacilitator protein and Z.mobilis glk-encoded glucose kinase in a host deficient in thephosphoenolpyruvate:carbohydrate phosphotransferase system.At10L scale withyeast extract supplementation,E.coli SP1.l pts/pSC6.090B synthesized87g/L ofshikimic acid in36%(mol/mol)yield with a maximum productivity of5.2g/L/h forshikimic acid synthesized during the exponential phase of growth.Shikimic acid(Figure1)is a naturally occurringhydroaromatic possessing a six-membered carbocyclicring arrayed with synthetically useful organic functional-ity and chirality.The utility of shikimic acid in chemicalsynthesis is reflected in its use as the starting materialfor the manufacture of Tamiflu,an orally effectiveantiinfluenza agent(1-4).Shikimic acid is also employedas a core scaffold in combinatorial library synthesis(5,6).More recently,reaction of shikimic acid in near-criticalwater was observed to produce phenol,thereby establish-ing a route for the synthesis of this large volumecommodity chemical from renewable feedstocks(7).Mi-crobe-catalyzed synthesis of shikimic acid from inexpen-sive glucose derived from abundant corn starch consti-tutes an alternative to the isolation of this hydroaromaticfrom relatively scarce Illicium plants(8,9).In thisaccount,the impact of increasing the availability ofphosphoenolpyruvate on the microbial synthesis ofshikimic acid and the scale-up of this synthesis from1to10L scale is examined.Condensation of phosphoenolpyruvate with D-erythrose4-phosphate catalyzed by3-deoxy-D-arabino-heptulosonicacid7-phosphate(DAHP)synthase(Figure1)is the firststep in the common pathway of aromatic amino acid biosynthesis that leads to shikimic acid.The in vivo availability of D-erythrose4-phosphate and phospho-enolpyruvate limit the catalytic activity of overexpressed, feedback-insensitive DAHP synthase in Escherichia coli K-12.Access to D-erythrose4-phosphate is increased upon overexpression of tktA-encoding transketolase(10,11). With improved D-erythrose4-phosphate availability, metabolic competition for phosphoenolpyruvate becomes a limiting factor in shikimic acid biosynthesis.The phosphoenolpyruvate:carbohydrate phosphotrans-ferase(PTS)system competes with DAHP synthase for in vivo supplies of phosphoenolpyruvate.PTS-catalyzed phosphoryl group transfer from phosphoenolpyruvate drives(Figure2)the transport of glucose into the microbial cytoplasm and phosphorylates glucose to form the glucose6-phosphate required for glycolysis(12).One molecule of phosphoenolpyruvate is converted into pyru-vic acid for each molecule of glucose transported into the cytoplasm.To evaluate the impact of increased phospho-enolpyruvate availability on shikimic acid biosynthesis, this study examined two basic strategies to contend with expenditure of phosphoenolpyruvate by the PTS system†Michigan State University.‡F.Hoffmann-La Roche,Ltd.Figure1.Biosynthesis of shikimic acid(SA).Intermediates and byproducts:phosphoenolpyruvate(PEP),D-erythrose4-phos-phate(E4P),3-deoxy-D-arabino-heptulosonic acid(DAH)7-phos-phate(DAHP),3-dehydroquinic acid(DHQ),3-dehydroshikimic acid(DHS),quinic acid(QA),inorganic phosphate(P i),nicotin-amide adenine dinucleotide phosphate(NADP).Enzymes: AroF FBR,DAHP synthase;AroB,DHQ synthase;AroD,DHQ dehydratase;AroE,shikimate dehydrogenase.808Biotechnol.Prog.2003,19,808−81410.1021/bp025769p CCC:$25.00©2003American Chemical Society and American Institute of Chemical EngineersPublished on Web04/04/2003in constructs cultured under fed-batch fermentor condi-tions at 1L scale.Cultivation of the construct synthesiz-ing shikimic acid in the highest concentration and yield was then examined at 10L scale.In one approach,pyruvic acid generated by PTS-mediated glucose transport was recycled to phospho-enolpyruvate (13,14)using amplified,ppsA -encoded phosphoenolpyruvate synthase (Figure 2).Phospho-enolpyruvate synthase converts pyruvic acid to phospho-enolpyruvate along with conversion of ATP to AMP and one molecule of inorganic phosphate (Figure 2).Expen-diture of phosphoenolpyruvate during glucose transport was avoided in a second approach by supplanting or augmenting PTS-mediated glucose transport with het-erologous expression of the Zymomonas mobilis glf -encoded (15-17)glucose facilitator (Figure 2).Transport of glucose into the cytoplasm by Glf-mediated facilitated diffusion does not expend either phosphoenolpyruvate or ATP.However,ATP is expended during the subsequent conversion of cytoplasmic glucose into glucose 6-phos-phate catalyzed by glk -encoded glucokinase (Figure 2).All shikimate-synthesizing E.coli constructs carried plasmid inserts that included aroF FBR -encoded,feedback-insensitive DAHP synthase and tktA -encoded trans-ketolase.Materials and MethodsGeneral.Culture medium and fermentation medium were prepared as previously described (9).Bacto yeast extract (15g/L)was added to the fermentation medium where indicated prior to adjustment to pH 7.0.All strains and plasmids used in this study are summarized in Table 1.Plasmid constructions were carried out in E.coli DH5R .Standard protocols were used for construction,purification,and analysis of plasmid DNA (18).T4DNA Ligase and Large Fragment of DNA Polymerase I were purchased from Invitrogen.Calf intestinal alkaline phos-phatase was purchased from Roche Molecular Biochemi-cals.PCR amplifications were carried out as described previously (18).Fed-batch fermentations at 1L scale were run under glucose-rich conditions and analyzed as previously de-scribed (9).A B.Braun Biostat ED fermentor (15L)connected to a DCU-2system was used for the fed-batch fermentations at 10L scale.The 10L fed-batch fermen-tations were run at 33°C.An impeller speed of no less than 300rpm was used to maintain dissolved oxygen (D.O.)levels at 30%air saturation.Addition of glucose (60%w/v)was initiated after the initial airflow rate of 0.2L/L/min was increased to 0.5L/L/min to reach the D.O.concentration of 30%of air saturation.To maintain a constant glucose concentration in the medium,the glucose feed rate was approximately equal to the glucose consumption rate as determined by either on-line mea-surements (GlucoTrace,Trace Biotech AG,Braunsch-weig,Germany),off-line measurements,or from the exhaust CO 2values based on an assumed 50%molar yield of CO 2per glucose added.SP1.1pts .P1phage-mediated transduction was em-ployed to transfer ∆(ptsH ptsI crr )::Km R from E.coli TP2811(20)into E.coli SP1.1(9).P1phage was propagated from TP2811,and transductions were carried out as described by Miller (21).Following the transduc-tion,cells selected from growth on LB medium containing Km R were screened for resistance to Tc and Cm.SP1.1pts possessed the following growth characteristics:growth on LB containing kanamycin (Km),tetracycline (Tc),and chloramphenicol (Cm);no growth on M9(9)medium supplemented with aromatic amino acids,aromatic vi-tamins,and L -serine;and light pink colonies on Mac-Conkey agar supplemented with 1%glucose.Transfor-mation of SP1.1pts with plasmid pSC6.090B afforded colonies that grew on M9medium supplemented with aromatic amino acids,aromatic vitamins and L -serine.pKD15.071B.This 11.9-kb plasmid was assembled by replacing the 1.0-kb -lac gene of pKD12.138(9)with the ppsA fragment from pKL1.87B.Plasmid pKL1.87B was digested with Bam HI and Hind III followed by treatment of the resulting 3.0-kb ppsA fragment with Klenow fragment.Following digestion of pKD12.138A with Nco I,the 8.9-kb fragment was modified to blunt ends using Klenow fragment.Ligation of these two purified frag-ments yielded pKD15.071B.pSC5.112B.To assemble this 11.1-kb plasmid,the 1.0-kb -lac gene of pKD12.138(9)was replaced with a glf -containing fragment obtained from pTC325(16).A 2.2-kb Bam HI/Hind III fragment containing the glf locus behind P tac was excised from pTC325and treated with Klenow fragment.Plasmid pKD12.138A was digested with Nco I and the resulting 8.9-kb fragment was treatedTable 1.Bacterial Strains and Plasmidsstrain/plasmid relevant characteristicsreference/source StrainDH5R lacZ ∆M15hsdR recAInvitrogen SP1.1RB791serA ::aroB aroL478::Tn 10aroK17::Cm R ref 9TP2811TP2111,∆(ptsH ptsI crr )::Km R ref 20SP1.1pts SP1.1∆(ptsH ptsI crr )PlasmidpSU18Cm R ,P lac lacZ ′,p15A repliconref 19pKD12.138Ap R ,aroF FBR ,tktA ,P tac aroE ,serA ref 9pTC325P tac glf glk ,lacI qref 16pKL1.87B Cm R ,ppsA in pSU18lab plasmid pKD15.071B ppsA ,aroF FBR ,tktA ,P tac aroE ,serA this study pSC5.112B P tac glf ,aroF FBR ,tktA ,P tac aroE ,serA this study pSC6.090BP tac glf glk ,aroF FBR ,tktA ,P tac aroE ,serAthis studyFigure 2.Transport of glucose into the E .coli cytoplasm.Enzymes:Glf,glucose facilitator;Glk,glucokinase;PTS,phos-phoenolpyruvate(PEP):carbohydrate phosphotransferase;PpsA,phosphoenolpyruvate (PEP)synthase.Biotechnol.Prog.,2003,Vol.19,No.3809本页已使用福昕阅读器进行编辑。