Determination of Phosphatidylcholine Content in Zophobas morio L. by High Performance Liqu
_HPLC-ELSD法测定前列地尔注射液中溶血磷脂酰胆碱的方法改进
表 1 梯度洗脱程序表 Tab 1 Gradient elution program
时间（ time） / min 0. 01 3. 00 3. 01 8. 00 8. 01
12. 00 12. 01 15. 00
流动相（ mobile phase）
A/%
B/%
25
75
50
50
100
0
100
0
0
100
本文采用改进后的 HPLC － ELSD 法测定前列 地尔注射液中溶血磷脂酰胆碱含量，采用梯度洗脱 的方法，将溶血磷脂酰胆碱与杂质完全分离，主成分 峰形理想，结果准确，能够有效地评价和控制制剂的 质量。 1 仪器与试药
仪器： Waters 2695 高 效 液 相 色 谱 仪，Waters 2420 蒸发光检测器。
0
100
25
75
25
75
取对照品溶液，进样 5 μL，按原标准方法及本 方法依法测定，色谱图见图 1。由图 1 可见，改进后 的方法获得的色谱图峰形良好，可用于溶血磷脂酰 胆碱的定性、定量分析。
图 1 原标准方法（ A） 与改进方法（ B） 溶血磷脂酰胆碱色谱峰形比较图 Fig 1 Comparison diagram of lysophosphatidylcholine chromatographic peak by the original standard method （ A） and the improved method （ B） 1． 溶血磷脂酰胆碱（ lysophosphatidycholine）
关键词： 前列地尔； 溶血磷脂酰胆碱； 杂质检测； 高效液相色谱； 蒸发光散射检测（ ELSD） ； 梯度洗脱； 标准改进
专业英语翻译作业
专业英语翻译作业Water is usually considered as being a compound of two elements.（我们）通常认为水是由两种元素组成的化合物Produced by electrons are the x-ray, which allow the doctor to took aside a patient’s body.x射线是由电子产生的,它能让医生看到病人的身体It is said that all matters is made up of atoms.（主语从句）据说，所有的物质都是由原子组成的。
Ancient people believed it to be true that the sun turned around the earth.（宾语从句）古代的人们相信太阳围绕地球旋转是千真万确的。
Why don’t we fall off the earth? The answer is t hat gravity keep s us from falling off.（表语）我们为什么不会从地球上掉下来？答案是因为有地球引力。
We are all familiar with the fact that nothing in nature will either start or stop moving of itself.（同位语）我们都熟悉这样一个事实：自然界中没有一个物体会自发开始运动或自发停止运动。
Fluids comprise both liquids and gases, the most common examples of which are water and air.（定语）流体包括液体和气体，最常见的例子是水和空气The body must dissipate heat as fast as it produces it.（状语）人体散发热量必须和产生热量一样快Up to the present time, throughout the eighteenth and nineteenth centuries, this new tendency placed the home in the immediate suburbs, but concentrated manufacturing activity, business relations, government, and pleasure in the centers of the cities.经历了18和19两个世纪直至今日，这种新的趋势是（人们）把住宅安排在城市的近郊，但把生产活动、商业往来、政府部门以及娱乐场所都集中在城市的中心地区。
HPLC_ELSD法测定大豆磷脂中磷脂酰胆碱和溶血磷脂酰胆碱含量_孙浩洋
为节省分析时间，本研究将长硅胶色谱柱和短硅胶色 谱柱分析结果进行比较（图 1 和图 2）。由图 1 和图 2 可看出，采用长硅胶柱时，PC 和 LPC 保留时间分别 为 25~30 min 和 53~70 min；而采用短硅胶色谱柱 AlltimaTM HP Silica（150 mm×4.6 mm，5 μm）时，PC 和 LPC 保留时间分别为 12~18 min 和 27~43 min， 相比长硅胶柱能大大缩短出峰时间，从而提高分析效 率。为进一步验证分析效果，将大豆改性磷脂用正己 烷―异丙醇（1 ∶ 2）溶液配成 10.0 mg/mL 样品溶液， 进行分析，色谱图见图 3。由图 3 可知，大豆改性磷脂 样品中 PC 和 LPC 可有效检出，并有很好分离效果。 故本研究选用短硅胶色谱柱（150 mm×4.6 mm，5 μm） 进行分析。
将 PC 标准品加入正己烷―异丙醇（1 ∶ 2）溶液 配制成浓度为 8.0 mg/mL、4.0 mg/mL、2.0 mg/mL、1.0 mg/mL、0.5 mg/mL 标准溶液。将 LPC 标准品加入正 己烷―异丙醇（1 ∶ 2）溶液配制成浓度为 6.0 mg/mL， 3.0 mg/mL、1.5 mg/mL、0.75 mg/mL、0.38 mg/mL标准 溶液。采用 1.3 中选择较优色谱柱及色谱条件下分析 PC 和 LPC 标准溶液；以峰面积对数为纵坐标，浓度 对数为横坐标，制作标准曲线。 1.4.2 样品 PC 和 LPC 分析测定
中图分类号：TQ645.9＋6
文献标识码：A
文章编号：1008― 9578（2012）05― 0025― 03
高效液相色谱_串联质谱法测定减肥保健食品中酚酞
90高效液相色谱-串联质谱法测定减肥保健食品中酚酞马微1,2，陈冬东3，王海波1，付丽4，代汉慧3，王秀君3，唐英章31.东宁出入境检验检疫局 (东宁 157200)；2.中国农业大学 (北京 100083)；3.中国检验检疫科学研究院 (北京 100123)；4.郑州牧业工程高等学校 (郑州 450011)摘要建立测定减肥保健食品中非法添加药物酚酞的高效液相色谱-串联质谱分析方法。
该方法对不同类型的减肥保健食品经超声波提取15 min，以Waters Atlantis T3柱(150 mm×2.1 mm, 3 μm)分离后，进行HPLC/MS/MS多反应监测模式下的定性及定量分析。
酚酞在(0.5~100) μg/L的范围内相关系数为0.999 4。
回收率71.3%~92.6%；日内精密度均小于8%，日间精密度均小于10%。
在检测的20种样品中，检出含有酚酞的样品9个，检出率为45%。
该方法分析速度快，灵敏度高，重现性好，可用于不同减肥保健食品中非法添加酚酞的快速检测。
关键词高效液相色谱-串联质谱；酚酞；质谱裂解途径；基质效应Determination of Phenolphthalein Added in the Weight-loss Functional Foods by Ultrasonication-high Performance Liquid Chromatography Tandem MassSpectrometryMa Wei 1’2，Chen Dong-dong 3，Wang Hai-bo 1，Fu Li 4，Dai Han-hui 3，Wang Xiu-jun 3，Tang Ying-zhang 31.Dongning Entry-Exist Inspection And Quarantine Bureau (Dongning 157200)；2.China Agriculture University, (Beijing 100083)；3.National Academy of inspection And Quarantine (Beijing 100123)；4.Zhengzhou College ofAnimal Husbandry Engineering (Zhengzhou 450011))Abstract To develop a comprehensive analytical method for the determination of phenolphthalein in the weight-loss functional foods by high performance liquid chromatography tandem mass spectrometry . Various weight-loss functional foods were extracted under ultrasonication in 15 min. Qualitative and quantitative analysis was carried out for the 20 samples under the MRM mode after the chromatographic separation on waters atlantis T3 (150 mm ×2.1 mm, 3 μm) column. Phenolphthalein was linear in the range (0.5~100) μg/L and the linear correlation coef fi cient reached 0.999 4. The mean recoveries were 71.3%~92.6%, with the intra-day precision less than 8% and the inter-day precision less than 10%. Phenolphthalein was measured in nine out of twenty samples. The detected ratio was 45%. The method is reliable, sensitive, reproducible and adapts to the determination of phenolphthalein in different weight-loss functional foods.Keywords high performance liquid chromatography tandem mass spectrometry ；phenolphthalein ；mass spectrum fragmentation pathways ；matrix effect 基金项目: 国家科技支撑计划农业领域课题（2006BAD27B02）酚酞（phenolphthalein）属于接触性轻泻药，口服后在肠内遇胆汁及碱性体液形成可溶性钠盐，刺激结肠黏膜促进肠蠕动，并可阻止肠液被肠壁吸收而起到缓泻作用[1]，所以常被非法添加到标榜为纯天然制剂的减肥保健食品中，使消费者服用后立即产生“少吃多排”现象，从而迅速减轻体重。
大豆磷脂的HLB值测定和乳化力的研究[1]
![大豆磷脂的HLB值测定和乳化力的研究[1]](https://img.taocdn.com/s3/m/201c62c3aa00b52acfc7cadd.png)
大豆磷脂的H L B 值测定和乳化力的研究安 红1,2,侯近龙1,邓启刚1,程侣柏2(1.齐齐哈尔大学化工学院,齐齐哈尔161006; 2.大连理工大学精细化工国家重点实验室)摘 要:建立了测定大豆混合磷脂的H LB 值的方法。
并做出了不同温度下H LB 值的标准曲线。
利用此曲线测定了不同PC 含量的磷脂的H LB 值,PC 含量为25.6%～69.9%,其H LB 值在7～10之间。
研究了不同PC 含量的大豆混合磷脂的乳化能力。
关键词:磷脂;H LB 值;磷脂酰胆碱;乳化力;水数法中图分类号:T Q545.1 文献标识码:A 文章编号:100124020(2003)1220716202ST UDY ON THE DETERMI NATI ON OF H LB VA LUE OF S OY BE ANPHOSPHATI DE AND THE E MU LSIFI ABI LITYAN H ong 1,2,H OU Jin 2long 1,DENG Q i 2gang 1,CHENG L übai 2(1.Institute o f Chemical Engineering ,Qiqihaer Univer sity ,Qiqihaer 161006,China2.State K ey Laboratory o f Fine Chemicals ,Dalian Univer sity o f Science and Engineering )Abstract :A feasible method (i.e the water titration method )for the determination of hydrophilic 2lipophilic balance(H LB )value of s oybean phosphatide was proposed.A calibration curve was prepared by titrating standard sam ples with different H LB values with water in a mixed s olvent of N ,N 2dimethylformamide (DMF )and benzene (DMF ∶C 6H 6=100∶15)s olution until a slight opaqueness is observed.In the preparation of calibration curve ,the standard H LB values were taken as the abscissa and the corresponding v olume (in ml )of the water titrated as the ordinate.In the analysis of unknown sam ple ,the H LB value was found from the calibration curve according to the v olume (in ml )of water used in the titration.The effect of tem perature during titration (20℃,25℃and 30℃)on the v olume of water used in the titration was als o studied in DMF 2C 6H 6s olutions containing different kinds of emulsifiers.Based on these findings ,it was found that the magnitude of H LB values of mixed phosphatides were related to the am ount of phosphatidylcholine (PC )in the mixture and to the tem perature of measurement.I t was als o found that when the PC contents were in the range of 25.6%～69.9%,the H LB values were varied in the range from 7to 10.Furtherm ore ,the emulsifi 2ability of mixed s oybean phosphatides containing different am ounts of PC was als o studied.K eyw ords :Water titration method ;Phosphatide ;H LB value ;Lecithin ;Emulsifiability H LB 值是表面活性剂的一个重要指标,表现了表面活性剂的亲油亲水平衡性。
HPLC法测定眼保_号胶囊中芍药苷的含量_谭家华
图22种化合物的清除DPPH自由基清除能力3讨论TBHQ是现今所用的合成抗氧剂中能力较强的一种，为了衡量芦荟药材中蒽醌类成分的抗氧化能力，将其清除自由基的作用于TBHQ清除自由基的作用进行比较，同时与常见的具有抗氧化能力的维生素E和维生素C进行了比较。
在浓度0 1mg/mL范围内，化合物的抗氧化性从大到小依次为羟基芦荟大黄素﹑维生素E﹑原儿茶酸﹑芦荟色苷G，其中在0 0.15mg/ mL范围内维生素C和TBHQ显示出强抗氧化性。
芦荟蒽醌类成分主要为芦荟大黄素苷及其苷元、芦荟大收稿日期：2012－01－05基金项目：贵州省中医药管理局中医药、民族医药科学技术课题研究项目（QZYY2010－28）作者简介：谭家华（1962－），女，贵州贵阳人，学士，副主任药师，主要从事中药质量控制与新药研究。
通讯作者：罗君（1980－），男，贵州遵义人，药师，硕士，主要从事中药质量控制与新药研究，E－mail：luojun031009@163．com。
黄素、芦荟苷、芦荟苦素、异芦荟苦素、6－羟基芦荟大黄素等。
蒽醌类化合物具有抗氧化和抗过氧化活性［4］，主要与蒽醌环上邻位或间位－OH有关。
对化合物8－C－glu－7－O－methyl－（S）–Al-oesol的鉴定工作还在进行中，还需做相应的图谱判断是否为新化合物。
本文通过制备芦荟中有代表性的化合物，对其进行简单的抗氧化能力的研究。
确定其应用价值。
其中，在DPPH自由基的抑制试验中，与常见的具抗氧化性的TBHQ﹑维生素C和维生素E进行比较，分析芦荟蒽醌类化合物可能在实际中的应用。
实验结果得出，在浓度范围0 1mg/mL内，羟基芦荟大黄素的抗氧化性强于原儿茶酸和芦荟色苷G，芦荟大黄素和8－C－glu－7－O－methyl－（S）－Aloesol不存在抗氧化性。
参考文献［1］邓军文．芦荟的化学成分及其药理作用［J］．佛山科学技术学院学报（自然科学版），2000，18（2）：76．［2］陈国和，刘玉鑫，张新申，等．芦荟的化学成分及其分离和分析［J］．化学研究与应用，2002，14（2）：46－48．［3］程旺兴．芦荟的化学成分及临床应用研究［J］．安徽医药，2001，5（1）：71．［4］袁阿兴，康书华，覃凌，等．斑纹芦荟的化学成分研究［J］．中草药，1994，25（7）：339－341．［5］王振月，宁洪斌．毛脉酸模根中大黄酚大黄素的分离鉴定［J］．中医药学报，1996，（2）：54．［6］陈屏，杨峻山．蒲葵籽化学成分的研究［J］．中国药学杂志，2008，43（21）：1669－1670．［7］肖志艳，陈迪华，斯建勇，等．库拉索芦荟化学成分的研究［J］．药学学报，2000，35（2）：120－123．HPLC法测定眼保Ⅰ号胶囊中芍药苷的含量谭家华，罗君，贺祝英，张建玲（贵阳中医学院第一附属医院，贵州贵阳550001）摘要：目的：建立HPLC测定眼保Ⅰ号胶囊中芍药苷的含量测定方法。
硫酸铵梯度法
挤压制备小粒径脂质体
• 通过挤压使脂质体通过固定孔径的滤膜，脂质体粒 径变小。该方法需要很低的压力，689kPa即可。
• 聚碳酸酯膜（polycarbonate membrane），该膜的通 道是直的并且大小相同，脂质体容易通过，即使脂 质体直径略大于孔径也能通过。一般将脂质体原液 稀释至12μmol/ml后再过膜，脂质体易通过孔径。
四、脂质体的表征
Characterization of liposomes
1．形态、粒径及其分布 2．包封率和载药量的测定
3．释放度与渗漏率的测定
4．药物体内分布的测定
思考题
1. 脂质体的研究与应用进展
2. 简述脂质体的概念、特点与用途。
3. 简述制备脂质体的常用的材料。
4. 简述pH梯度法制备载药脂质体的原理和方法。
2．负电荷磷脂 负电荷磷脂又称为酸性磷脂 • 磷脂酸 phosphatidic acid, PA
• 磷脂酰甘油 phosphatidyl glycerol, PG
• 磷脂酰肌醇 phosphatidylinositol, PI
• 磷脂酰丝氨酸 phosphatidyl serine,PS
3．正电荷脂质 均为人工合成产品
• 脂质体加压通过孔时，其结构发生变化，根据所需 脂质体的大小选择膜的孔径。
3．主动包封法 Remote loading method
• 从根本上改变了难以制备高包封率脂质体的局面。
• 主动包封技术的应用与药物的结构密切相关，不能 推广到任意结构的药物，因而受到了限制。
• 弱碱性的药物
采用pH梯度法、硫酸铵梯度法
2．逆相蒸发法 Reverse-phase evaporation
由Szoka提出
脂质体电动色谱用于评价药物透皮吸收
史上最快最全的网络文档批量下载批量上传，尽在：/item.htm?id=9176907081 史上最快最全的网络文档批量下载批量上传，尽在：/item.htm?id=9176907081脂质体电动色谱用于评价药物透皮吸收王永军，刘洪卓，孙进，孙英华，刘晓红，何仲贵※(沈阳药科大学药学院生物药剂学研究室，辽宁沈阳，110016)摘要：目的采用脂质体电动色谱法对药物透皮吸收进行评估。
方法测定22个不同种类化合物的保留因子(k)，采用逐步回归法在药物透皮参数(logKp)和log k及其它简单理化参数之间建立定量保留活性关系(quantitative retention-activity relationships, QRARs)。
结果建立了药物透皮参数(logKp)和log k、分子量、化合物OH、NH键数目之和之间的相关方程，相关系数R2 = 0.902。
结论脂质体电动色谱在预测药物透皮系数方面展现了良好的应用前景。
关键词：脂质体；电动色谱；透皮参数；逐步回归Predicting skin permeability using liposome electrokinetic chromatogrpphyYongjun Wang, Hongzhuo Liu, Jin Sun, Yinghua Sun, Xiaohong Liu, Zhonggui He※ABSTRACT: OBJECTIVE To estimate drug penetration through skin using liposome electrokinetic chromatography. METHODS The retention factor of twenty-two structure diversely compounds were determined, quantitative retention-activity relationships were constructed between skin permeation coefficients and logk and other physicochemical parameters by stepwise regression method. RESULTS a quantitative retention-activity relationships (QRARs) was constructed between compound skin permeability coefficient (logKp) and log k, molecular weight, the counts of OH and NH of molecule by stepwise regression method (R2= 0.902). CONCLUSIONS In a word, LEKC is a promising rapid tool to predict drug penetration throughskin.课题获国家自然科学基金No. 30400563和辽宁省自然科学基金No. 20052058资助。
气相色谱-质谱联用法测定植物油中脂肪酸组成
气相色谱-质谱联用法测定植物油中脂肪酸组成邓莉!，何静仁，何毅，舒静（武汉轻工大学食品科学与工程学院，湖北省农产品加工与转化重点实验室，武汉430023）摘要：建立气相色谱-质谱联用法测定植物油中脂肪酸组成的方法。
选用硼化氟-甲醇体系对样品进行甲酯化处理，再通过气相色谱-质谱技术对脂肪酸组成进行分析和鉴定$共分离出11种脂肪酸，结果表明：植物油中的脂肪酸以油酸、亚油酸、棕桐酸、硬脂酸为主，其中油酸和亚油酸含量均达70%以上，不饱和脂肪酸含量与饱和脂肪酸含量的比值：玉米油5.9%，菜籽油17.6%，葵花籽油11.7%，花生油3%，调和油5%$植物油中不饱和脂肪酸含量远远高于饱和脂肪酸含量$关键词：气相色谱-质谱联用；植物油；脂肪酸中图分类号:TS225.1文献标志码：A doi：10.3969/j.issn.1000-9973.2019.06.036文章编号：1000-9973(2019)06-0157-03Determination of Fa#y Acid Composition in Vegetable Oilby Gas Chromatography-Mass SpectrometryDENG Li*,HE Jingren,HE Yi,SHU Jing(Key Laboratory of Processing and Conversion of Agricultural Products in Hubei Province,College of Food Science and Engineering,Wuhan Polytechnic University,Wuhan430023,China) Abstract：A method for determination of fatty acids in vegetable oil is developed by gas chromatography-mass spectrometry(GC-MS).Boride fluorine-methanol system is used to methylate the samples,and then the fatty acid composition is analyzed and identified by GC-MS.11kinds of fatty acids are identified. The results show that the main fatty acids are oleic acid,linoleic acid,palmitic acid,stearic acid,and the content of oleic acid and linoleic acid is more than70%,the ratio of unsaturated fatty acid content to saturated fatty acid content is 5.9%corn oil,17.6/rapeseed oil,11.7/sunflower oil,3/ peanut4iland5%blended4il.Thec4ntent4funsaturatedfa t yacidinvegetable4ilis muchhigher than that of saturated fatty acid.Key words：gas chromatography-mass spectrometry(GC-MS)；vegetable oil；fatty acid植物油是从植物的果实、胚芽和种子中提取得到的一种可食用油脂,是人们日常饮食的重要组成部分（12）。
电子自旋标记于两种共存的磷脂质相态中的不对称分布
电子自旋标记于两种共存的磷脂质相态中的不对称分布赖奕丞;江昀纬【摘要】该文探讨自旋标记于不同磷脂质相中不对称分布的分子机制.透过电子自旋共振(ESR)波谱技术以及理论波谱分析, 探究两种在磷脂质不同烷基位点上的自旋标记(16-PC & 5-PC)于两种明显不同物理性质的磷脂质 (DPPC & DLPC) 所构成的细胞膜两相共存区域具有不对称分布情形.根据实验结果, 如果自旋探针平均分布在两相之间, 则其波谱将能根据杠杆法则(lever rule) 来拟合波谱的结果, 而自旋探针的分布反映了该探针周围环境的脂质所给予的立体障碍, 而这个不对称的情形可以合理的解释成自旋标记的表面自由面积所造成.根据结果, 16-PC会平均分布在两个不同的相区域, 而5-PC则会有不对称分布的情形.【期刊名称】《波谱学杂志》【年(卷),期】2010(027)003【总页数】15页(P470-484)【关键词】电子自旋共振;分子有续参数;杠杆法则;相共存【作者】赖奕丞;江昀纬【作者单位】清华大学,化学系,台湾,新竹,30013;清华大学,化学系,台湾,新竹,30013【正文语种】中文【中图分类】O482.53IntroductionNitroxide spin label has been extensively used to determine the phase separations in lipid mixtures[1-4]. These diagrams indicated lateral phase separations of lipids into do mains being different compositions, molecular ordering, and fluidity, based on the quantitative descriptions derived from the spectral analysis of electron spin resonance (ESR) data. Thus far, however, the asymmetric partitioning of spin label has been rarely discussed.In membrane biophysics, partitioning of probe molecules into the various thermodynamic phases has been an important issue, because membrane is characterized by complex structures, which are largely heterogeneous[5]. Ge et al.[6] used five spin labels (5-PC, 7-PC, 10-PC, 16-PC and spin-labeled cholestane) to study the phase coexistence of plasma membrane vesicles prepared from RBL-2H3 mast cells. Moreover, spin label approach has also been used to investigate the coexistence of membrane phases in five different cell lines[7]. It was found that in the coexistence phases of liquid-ordered and liquid-disordered the partitioning of different spin label varies with respect to the attached position of spin label on the lipid acyl chains. Since lipids consist of acyl-chains and polar head moieties, their physico-chemical properties are dependent upon the length and degree of unsaturations of acyl-chains, and a variety of polar heads[8, 9]. Similar results were also observed in 250 GHz high-field ESR application[10]. Thus selection of probe molecule is the most crucial matter in the dynamicstudies of biomembranes. The phase fraction in a multi-phase coexisting region would never be correctly studied unless the partitioning coefficient of spin label is adequately evaluated in advance.Interactions between membranes and polypeptides or any solute molecule inserting into membranes are a central issue regarding the insertion and folding of peptides in membrane. Although the mechanism of protein-membrane interactions involves numerous processes, they all must undergo, at an early stage, solutes partitioning between water and lipids and among coexistence phases in membranes. Some rigorous discussions on chain packing theories for micelles and bilayers can be found in early 1980s[11, 12]. Since then, many concerns have been raised that the lipid bilayers and vesicles have been commonly, but incorrectly, viewed as a slab of bulk hydrocarbon liquid and treated with solubility-diffusion theory[13-16]. It was found that the partitioning of solutes between water and lipid membranes is affected by the free-surface-area at which solutes reside in the lipid membranes, as well as the size differences of solutes and membrane molecules. In studying various model phospholipids (DPPC, DLPC, DMPC and n-alkanes) with molecular dynamics simulations[14, 17-21] and experimental measurements[15, 16, 21, 22], it was demonstrated that the partitioning of solutes into membranes is molecule-size dependent requiring the Flory-Huggins theory to correct the value of transfer free energies of solutes, which is originally calculated from general solubility-diffusion model. In other words, the partitioning of solutes is affected by the local environment where it resides at, and the molecularsize differences of solutes and solvents. This issue has not been discussed in the field of the spin-label ESR.In the present report, we describe the asymmetric partitioning of the spin labels, 5-PC and 16-PC, found in lipid phases. This finding is to be manifested by ESR spectral simulations and further confirmed by linear combination of ESR experimental spectra, which is based on the lever rule, a thermodynamics principle. Relevant data for the asymmetric partitioning of 16-PC was previously reported[4, 7]. However, the mechanism of the spin labels partitioning into lipid membranes has never been discussed. Our aims therefore are to i) report the asymmetric partitioning of 5-PC and 16-PC spin labels in the DPPC-rich (Lβ) and DLPC-rich (Lα) coexistence phases and ii) examine the feasibility of applying the existing theories, which were originally used to study solutes partitioning between water and lipid bilayers, to our current system of two-phase coexistence.1 Experimental section1.1 MaterialsPhospholipids and spin labels, 1-palmitoyl-2- (16-doxyl stearoyl) phosphatidylcholine (16-PC) and 1-palmitoyl-2-(5-doxyl stearoyl) phosphatidylcholine (5-PC) were purchased from Avanti Polar Lipids, Inc. (Alabaster, AL). All materials were used without further purification.1.2 Sample preparation and ESR spectroscopyThe sample preparation was previously described in detail. In particular, the spectra of 16-PC were investigated previously to determine the phase diagram of ternary phase diagram[4]. It is briefly summarized below. Asystem of binary lipid mixtures, DPPC and DLPC, was studied. Measured stock solutions of the lipids, and 16-PC and 5-PC in chloroform were mixed in a glass tube. The concentration of spin label was 0.25 mol% of the lipids. The ratio of lipid composition was varied in the range of χDPPC=0.0～1.0. Multibilayer samples were prepared by the method of Rapid Solvent Exchange[23] in a buffer of Pipes/KCl/EDTA=5 mmolL-1/200 mmolL-1/1 mmolL-1 at pH 7.0. Samples were then pelleted using a desktop centrifuge, and transferred to microcapillaries with excess buffer and sealed with paraffin.ESR spectra were obtained on a Bruker Instruments EMX ESR spectrometer at a frequency of 9.55 GHz at a room temperature. In the present study, the recorded ESR spectra were normalized with the double integration of a first-derivative spectrum method and aligned with central peak in prior to the fitting procedure.1.3 ESR spectral simulationsNonlinear least-squares (NLLS) fitting program based on the stochastic Liouville equation (SLE) was used to analyze the spectra from spin labels attached to the acyl chains of the lipids[24]. The dynamic parameters used in the fitting program are rotational diffusion rates (R⊥) and the order parameter (S0) whose molecular axis systems and definitions have been well defined for spectral simulations of membranes[9]. The MOMD model, which stands for microscopic order and macroscopic disorder, was used in the simulation[25] .1.4 Determination of partition coefficient by linear combination of ESRspectraThe bottom line (χChol=0%) of the DPPC-DLPC-Cholesterol ternary phase diagram is known as a tie line[4, 26]. The compositions of the components along the tie line can be predicted from lever rule, which is equivalently, to construct the spectra of the samples in the two-phase coexistence phase by linearly combining two basis spectra on the boundaries[26]. The underlying idea of using linear combination of ESR experimental spectra to determine the partition coefficient Kp is based on Eq. (1) and (2), where vectors A and B represent the two basis set of spectra at a boundary.Here γ represents the component ratio that best fit the samples in the two-phase coexistence region. The spectrum, which is simulated by linear combining the two boundary spectra, is represented as the vector Φ, shown in Eq. (1). The partition coefficient Kp is given by Eq. (2), where μ is a prediction ratio of A component by the lever rule, and γ is the best-fit ratio of A component. The γ value can be determined either by spectral simulation or linear combination of experimental spectra, providing a comparison for the simulations and experiments. Details are given in Result. In other word, the first part of Eq. (2) is the mole percentage ratio of the phases, which is predicted by the lever rule. The second part is the mole faction ratio of the spin label being in each phase. If the spin label partitions into each coexisting phase equally Kp yields unity; whereas, if Kp value is greater than unity it indicates spin label favors of the phase of A component. In principle, Kp values obtained from fitting samples along atie line must be an invariant. Based on this criterion, we may combine Eq.(1) and (2) to obtain Eq. (3).(3)We then performed the least squares fitting procedure to determine Kp values from ESR experimental spectra. Since the vector Ф is the combination of spectra A and B, the problem is then to find the Kp that best fits Φ(Kp) with the experimental ESR spectra according to the minimum in the squared differences or χ2 values between the data. Note that the underlying principle of the procedure can be further applied to determine tie-lines from trail lines[26].2 ResultsFig.1 shows the phase diagram of a lipid mixture of DPPC and DLPC as determined by fluorescence spectroscopy[5] and ESR measurements[4] that t here are two phase boundaries locating at χDPPC=0.3 and 0.85, dividing the system into three regions, which are DLPC-rich liquid-disordered phase (Lα) region, DPPC-rich gel phase (Lβ) region, and a region where DLPC-rich and DPPC-rich phases coexist. To reveal the partitioning parameter of a spin label in the two-phase coexisting region, we first fit the spectra in one phase region to obtain two sets of parameters locating in the nearest neighborhood of two boundaries, i.e., the samples at χDPPC=0.3 and 0.85. Based on the thermodynamic principle that the physical property of each component is an invariant along a tie line, dynamic parameters of each boundary component werefixed while the component ratio was varying in the simulations for the spectra within two-phase region.Phase diagram of DPPC-DLPC lipid mixturesFig.1 The phase diagram of DPPC and DLPC mixtures (χDPPC+χDLPC=1.0). The region χDPPC=0～0.3 is Lα phase, which is the DLPC-rich fluid lamellar phase, and the region χDPPC=0.85～1.0 is Lβ phase, whi ch is the DPPC-rich ordered phase. These two phases (Lα and Lβ) coexist in the regionχDPPC=0.35～0.82.1 Dynamic parameters in the Lα and Lβ coexistence phaseThe ESR spectral simulations of 5-PC and 16-PC from the well-defined coexistence phase, DPPC-rich and DLPC-rich phases, were performed using the NLLS fitting program. We first analyze the boundary spectra using NLLS. The 5-PC spin label intrinsically has more constraint on its motion than does the 16-PC. This was manifested by our simulation results, which show that the axial rotational diffusion rate R⊥ of 16-PC is greater than that of 5-PC while the S0 of 16-PC is less than that of 5-PC. The R⊥ and S0 for 16-PC in the DPPC-rich component are 7×107 s-1 and 0.15, respectively, while those for 5-PC in the same phase component are4.1×107 s-1 and 0.38, respectively. This is consistent with the general consensus that the 16-PC has less constraint on the local environment than 5-PC due to the different position on the acyl chain. Same observations were obtained for the other coexisting component, DLPC-rich, that the R⊥ and S0 for 16-PC are 17.7×107 s-1 and 0.02, respectively; while those for 5-PC are5.9×107 s-1 and 0.29. These dynamic parametersare consistent with the general assignments of Lα and Lβ phase s.Fig.2(a) shows the fitting results for the ESR spectra from 16-PC in the DPPC-rich and DLPC-rich coexistence region, where the solid and dashed lines represent the experimental and simulated spectra, respectively. The spectra in the region were simulated to fit the experimental data by two seed spectra of lipid vesicles right outside the two-phase coexistence region. The component ratio was varied to fit the experimental spectra, while the dynamic parameters were fixed in the two-phase coexistence region. NLLS fitting program returns the best fitting spectra with minimal chi squared value, χ2. Fig.2(b) shows the fitting results of 5-PC in the DPPC-rich and DLPC-rich coexistence region. All fitting procedures are the same as those of 16-PC.Fig.2 ESR spectra from (a) 16-PC and (b) 5-PC in multilamellar vesicle of DPPC and DLPC mixtures at room temperature. The mole fraction of DPPC (χDPPC) is indicated. Solid and dashed lines represent experimental data and simulations, respectively2.2 Partition coefficients by ESR spectral simulationsTo estimate the partitioning condition of different spin label in the two-phase coexistence region, the partition coefficient Kp, defined in Eq. (2), is used to reveal the ratio of the two coexisting phases (Lβ & Lα). The uni ty of Kp indicates the equal partition of the applied spin label. In the present report, Kp>1 denotes that spin label favors of Lα phase. The estimated values of Kp for 5-PC and 16-PC in the two-phase coexistence region are summarized in Table 1. The first column represents the componentpercentage of DPPC in the mixture of DPPC and DLPC. The second column indicates the phase percentage of Lα, predicted from lever rule, in the Lα and Lβ coexistence region. The simulation results are shown in the succeeding columns. The average Kp values for 5-PC and 16-PC are 3.65 and 0.88, respectively; the standard deviations (σ) of which are ca. 1.63 and 0.24.Fig.3 shows correlation plots for 16-PC and 5-PC in the DPPC-rich and DLPC-rich coexistence region. According to the lever rule, the theoretical correlation plot yields a linear diagonal line (solid line with asterisk points), assuming the symmetric partition of the probe molecule. The simulation data derived from the 16-PC spectra (open square) were distributed in the vicinity of the line predicted by the lever rule. The average Kp accounts for more exactly that the 16-PC partitions in DPPC-rich and DLPC-rich phases with a ratio of 1∶0.9. The asymmetric partitioning of 5-PC in the coexistence region is clearly displayed in the correlation plot (Fig.3). The data of 5-PC distributed far from the line of equal partition and yielded the partitioning ratio of DPPC-rich to DLPC-rich by 1∶3.6 ratio.Table 1 The partition coefficients (Kp), determined from ESR simulations, for 5-PC and 16-PC as incorporated in DPPC/DLPC multilamellar vesiclesχDPPCDPPC-rich on the Tie-line (%)5-PC16-PCDPPC-rich (%) (simulation)KpDPPC-rich (%)(simulation)Kp0.890.973.13.7093.30.720.7581.857.63.3386.20.730.772.749.7 2.7074.40.920.6563.634.63.3364.60.960.654.515.16.6762.50.720.5545.5--47.30.930.536.411.14.5440.80.830.4527.38.74.0020.51.450.418.219.40.93--0.359.1--13.60.64Avg:3.65σ:1.63Avg:0.88σ:0.24Fig.3 The correlation plot between the partition ratios for 16-PC (open square) and 5-PC (open circle) in the DPPC-rich and DLPC-rich coexisting region. Sold line with asterisk represents the lever rule predictions with an equal partitioning of the spin labels. The respective average partition coefficients for 16-PC and 5-PC are ca. 0.88 and 3.652.3 Partition coefficients determined by the spectral linear combination methodThe analysis results of the partition coefficients of 16-PC and 5-PC using linear combination of experimental spectra are summarized below. Each spectrum in the two-phase region was fit with a Kp. Thus, the Kp and the standard deviation σKp is shown in average sense instead of using one single Kp to fit all spectra along a tie line. Theoretically, Kp must remain invariant along a tie line. This is the original idea proposed for the determination of partition coefficients using spectral combination. However, we found that fitting all spectra with a single Kp is impractical since the inaccuracy is always promoted by un-anticipated experimental errors. In other words, tiny experimental inac curacy will blur the χ2 values of hypothetical lines and increase ambiguity in χ2 values. Therefore, by fitting each spectrum with one single Kp and observing the result of σKp, we can take the error bar for the Kp values into consideration and exclude some spectra, which result in abnormal values of Kp, from the analysis. The partition coefficients for 16-PC and 5-PC were, therefore, finally determined to be 0.92 and 3.66, respectively. The respective standarddeviations are 0.35 and 1.28. Two abnormal values were observed(Kp<0.01 for χDPPC=0.8 in the 16-PC series and Kp>15 for χDPPC=0.6 in the 5-PC series) and, therefore, excluded from the analysis. These abnormal deviations of Kp were mainly caused by the fluctuations in experimental preparations. The partition coefficients determined by spectral combination (Kp,16-PC=0.92 and Kp, 5-PC=3.66) are well consistent with those obtained from NLLS spectral simulations (Kp, 16-PC=0.88 and Kp, 5-PC=3.65, shown in Table 1).3 Discussion3.1 Fluorescent spectroscopic evidences of asymmetric partitioning Asymmetric partitioning has been reported for various fluorescent probes[27]. Fluorescence quenching measurements on indolyl-labeled lipids have shown that indole ring as attached to 16 acyl-chain cannot partition into the lipid membranes. However, the 18 acyl-chain with indole can partition into the lipid membranes[28, 29]. Using the fluorescence resonance energy transfer (FRET) method, an asymmetric partitioning has been found in the fluorescent probes located in the gel, fluid phases, and the gel/fluid interphase[30]. It was found that a fluorescent dye molecule with acyl chains of 12 carbons prefers a fluid phase, i.e., Lα phase, whereas a dye molecule with 18 carbon acyl chains mainly partitions to the Lβ gel-phase. The study of solubility of two fluorescent lipid amphiphiles with comparable apolar structures and different polar head groups, NBDhexadecylamine and RG-tetradecylamine (or -octadecylamine), shows that short, fully saturated chains (C12:0) and long chains with a c/s-unsaturation partitioned preferentially into disordered phase domains; whereas long fully saturated chains showed no phase preference[31]. The periphery of acyl chains is more randomly distributed in different phases. These findings show some relevance with the asymmetric partitioning of 5-PC in the present report. It infers that asymmetry may exist in the partition coefficients of lipid spin labels.3.2 Molecular basis of the asymmetric partitioningOur spectral simulation and experimental data combination results indicated that 5-PC prefers to DLPC-rich phase, which is characterized by higher fluidity in acyl chains than in DPPC-rich phase, with a ration of3.66∶1. On the other hand, 16-PC has (almost) no preference between the two coexistence phases. The hydrocarbon chains in such amphiphilic mesophases, such as lipid bilayers or multilamellar membranes vesicle, are highly ordered near the interfacial plane of the polar head groups. The order of acyl chains decreases with distance from the interface. This is the so-called gradient of disorder and has been confirmed by studying the packing of short-chain (15-carbon fatty acid chain) molecules in monolayers and bilayer membranes using the theory of lattice model[18] and the theoretical simulations of molecular dynamics and statistical mechanics for lipid membranes[32, 33]. The lattice model was further used to show that the configurations of the hydrocarbon chains in micelles and vesicles are severely constrained by the space-filling requirements of the chain segments and by the continuity of the chains[19]. The gradient of disordering was thus rationalized in terms of surface density and chainlength of the phospholipids. A question raised from these theoretical studies was whether the solubility-diffusion free energy (ΔG=-RTlnKm, where Km is the mole fraction ratio of solute in bulk phase), used for studying solute partitioning between bulk phases, could possibly fail in predicting solutes partitioning among lipid membrane phases. This issue was not clarified until De Young and Dill measured the membrane/water partition coefficients of benzene into lipid bilayers (DLPC, DPPC and DMPC) as a function of the surface density of the phospholipid chains using 2H NMR. It showed that i) increasing surface density of phospholipids chains dramatically leads to solute exclusion; ii) the partitioning of solutes into bilayer membranes is of a fundamentally different nature from partitioning into bulk phases[15]. These findings are consistent with the ESR studies showing that the ordering of spin labels decreases with increased chain length of attached spin labels[9, 10]. Thus the solute uptake increases significantly with the increased chain length of lipid bilayers. In order to determine the relative contributions to partitioning from chain ordering and surface density, the partition coefficients for benzene between an aqueous solvent and phospholipids bilayer membranes were measured as a function of the surface density of the bilayer chains. It has been found that the partition coefficients for benzene between water and n-alkanes (from C8 to C16) were independent of alkane length only if Flory-Huggins (FH) theory, which is more appropriate for a polymer-like system, was used to correct for the molecular size differences of solutes and solvents[16]. This provided supporting evidence for that i) the molecular size (solventand solute) affects mole-fraction-based solubility-diffusion free energies between chain hydrocarbons and water; ii) with the application of FH theory that corrects free energy of solute with solute volume unit, the free energy that determines the partitioning would become independent of chain length. In addition of experimental measurements, there have also been a several theoretical studies. Based upon mean-field model and Monte Carlo simulations, FH theory has been proven useful for evaluating the partitioning for solutes of arbitrary shapes, as long as the solvent is chain-like[17, 20, 34].The same conclusion, that the permeability of solutes across lipid bilayers is dependent on lipid chain packing, was also drawn by studying the Lα and the Lβ phases of DPPC bilayers (with cholesterol variation). This was confirmed by NMR and dynamic light scattering studies[22, 35]. These studies addressed that a scaling factor must be applied to the predictions from solubility-diffusion theory to correct for chain ordering from NMR data. Based on experiments of a variety of membranes, including DLPC, DMPC, DPPC and DSPC, with varying cholesterol concentration and temperature using seven short-chain monocarboxylic acids as permeants, they have satisfactorily explained the chain-length-dependent partitioning of solutes with a free-surface-area theory using FH theory.The most commonly employed model, solubility-diffusions model, for describing the passive transport of permeants across different bulk phases is thus inappropriate for manifesting the chain-length-dependent partitioning of solutes found in the lipid membranes studies. The FHtheory corrects the transfer free energy calculated from solubility-diffusions model by introducing a molecular-size-based volume ratio, so that the volume-ratio-based free energy is independent of chain length after the correction. Similarly, the free-surface-area theory is to account for the decrease in permeability coefficient, which is predicted from a bulk solubility-diffusion model, by introducing a scaling factor for the permeability decrement term. The FH theory and the free-surface-area theory provided the most reliable rationale for explaining the results of the present report that i) the portioning of the spin labels into different phases is dependent on the local free-surface-area where the spin label is attached on the hydrocarbon chains; ii) the partitioning dramatically decreases with decreased free-surface-area of the local environment, which is quantitatively consistent with the trend of the order parameter variation extracted from ESR spectral simulations.3.3 Relation between the free-surface-area and the order parameterMost of the previous studies of investigating solutes partitioning into membranes were based on observation of the surface density variations extracted from 2H NMR studies. The surface density is defined as A0/A, where A is the area occupied per phospholipids molecule and A0 is the area in the crystal. This surface density varies from 0 to 1; the value of 1 represents the completely ordered crystalline state. In the experiments of hexane partitioning into n-alkanes solution, they have observed hexane partitioning decreases by a factor of 9 as the surface density of the bilayer chains increase from 0.5 to 0.9[16]. Similar results have been found inmany studies of membrane systems[15, 22, 35]. They have shown that partitioning increases dramatically in the range from 0.5 to 0.9 of surface density; whereas partitioning is almost independent of solute molecular size and local free-surface-area of acyl chains as the surface density decreases below 0.5[15, 16, 22]. These results are consistent with the present study in terms of order parameter; i.e. the partition coefficient for 5-PC is over 3 (ratio of Lα to Lβ) as order parameter of Lα and Lβ phases is 0.29 and 0.38 respectively; whereas, the partition coefficient of 16-PC is about unity although the order parameter is 0.02 in Lα phase and 0.15 in Lβ phase. This indicated that the local free-surface-area of 16-PC either in Lα or Lβ phases has been large enough to accommodate a nitroxide spin label independently of the solute molecule size, while the local environment of 5-PC, however, is still being in high surface density (high ordering) and has great influence on partitioning of spin labels. This factor plays an important role in preventing 5-PC from partitioning into Lβ phase, resulting in the partition co efficient of 3.66 with favoring of Lα phase. Fig.4 Order parameters Smol (for 2H NMR) and S0 (for ESR) as a function of the labeled position on lipid acyl chain. Open triangle (△) and open circle (○) are 2H NMR data of Seelig[36], and represents POPC and D PPC being in Lα phase respectively. Square represents S0 obtained from ESR simulation in Lα phase (open square) and in Lβ phase (shaded square) respectively. The surface density converted from S0 is shown in parenthesesA comparison between the order parameters of various positions on acylchain obtained from 2H NMR (Smol) and CW-ESR (S0) is shown in Fig.4. The open triangle and open circle are order parameters obtained from 2H NMR[36], and represent POPC and DPPC in Lα phase, respectively. The open and shaded square boxes are order parameters obtained in the present study, and represent spin labels being in Lα and Lβ phases, respectively. The trends of order parameter variations from NMR and ESR data are consistent with each other. The numbers shown in parentheses in Fig.4 are the surface density (σS) converted from ESR order parameters using Eq.(4), which was originally derived in NMR[15]. The partitioning is dramatically decreased by a factor of 3.66 as the surface density is varied from 0.53 in Lα phase to 0.59 in Lβ phase, while it is hardly changed as the surface density is below 0.5 either in Lα phase or Lβ phase. This shows the partitioning of spin label is influenced by surface density as the same way as that for 2H NMR studies. Note that there is no direct conversion between the order parameters obtained from 2H NMR and ESR data, although they possess the same physical meaning in terms of angular fluctuations of chain segments. The main reason for the difference is due to operating frequencies in NMR and ESR experiments, resulting in detecting different time windows of the molecular dynamic.(4)3.4 ESR simulation method for the determination of the asymmetric partitionThe simulations were carried out to evaluate dynamic parameters, R⊥ and。
吩噻嗪类衍生物
1. Introduction Phenothiazines are important psychotropic compounds, but they also have further biological activities.1–3 For example, phenothiazines have recently been considered as potential drugs in the management of CreutzfeldtJacob disease.4 Metabolism of phenothiazine-based drugs often results in the formation of 7-hydroxylated derivatives or 5-sulfoxides.5–7 Because oxidation of asymmetrically substituted phenothiazines at the S(5) position introduces a new stereogenic center, these 5-sulfoxides are chiral. Although chiral 5-sulfoxide metabolites of the phenothiazine drug thioridazine in human plasma were separated by HPLC,8 to date optically active phenothiazine 5-oxides have not been obtained on preparative scale. Hence, stereoselective methods for the synthesis of optically active phenothiazine-5-oxides would extend the possibilities for investigation of the S -oxide metabolites of phenothiazine-based drugs.
211018619_HPLC_法测定前列地尔注射液中溶血磷脂酰胆碱和溶血磷脂酰乙醇胺的含量
第52卷第3期 辽 宁 化 工 Vol.52，No. 3 2023年3月 Liaoning Chemical Industry March，2023收稿日期： 2022-08-04 作者简介： 张硕（1987-），女，满族，辽宁省沈阳市人，主管药师，2011年毕业于沈阳药科大学药学专业，研究方向：药品研发。
HPLC 法测定前列地尔注射液中溶血磷脂酰胆碱和溶血磷脂酰乙醇胺的含量张硕1，付永慧2，武立华1，孙银玲1，陈阳1（1. 远大生命科学（辽宁）有限公司，辽宁 沈阳 110171; 2. 蓬莱诺康药业有限公司，山东 蓬莱 265600）摘 要：研究了HPLC -ELSD 法测定前列地尔注射液中溶血磷脂酰胆碱和溶血磷脂酰乙醇胺的含量。
采用低温型蒸发光散射检测器（雾化气为氮气，雾化气压力为 25 psi，漂移管温度为40 ℃），Ultimate Diol 色谱柱（4.6 mm×250 mm, 5 μm ）；以甲醇-水-冰醋酸-三乙胺（85∶15∶0.5∶0.05）为流动相A，以正己烷-异丙醇-流动相A（20∶48∶32）为流动相B，柱温为40 ℃，梯度洗脱。
结果表明：溶血磷脂酰胆碱在0.02～0.20 mg ·mL -1（R =0.999 9，n =6），溶血磷脂酰乙醇胺在0.01～0. 1 mg ·mL -1 （r =0.999 8，n =6）成良好线性关系；定量限分别0.02 mg ·mL -1和0.01 mg ·mL -1，检测限分别为0.006 mg ·mL -1和 0.003 mg ·mL -1；平均回收率（n =9）分别为101.4%（RSD =2.4%）和98.8%（RSD =1.9%）。
所建立的HPLC 方法可用于前列地尔注射液中溶血磷脂酰胆碱和溶血磷脂酰乙醇胺含量的测定。
关 键 词：HPLC；前列地尔注射液；溶血磷脂酰胆碱；溶血磷脂酰乙醇胺；含量 中图分类号：O657 文献标识码： A 文章编号： 1004-0935（2023）03-0457-04前列地尔，又叫前列腺素E1，化学名 (1R, 2R, 3R)-3-羟基-2(E)-(3S)-3-羟基-1-辛烯基-5-氧代环戊烷庚酸，是一种前列腺素。
磷酸果糖激酶（PFK）活性检测试剂盒说明书
磷酸果糖激酶（PFK ）活性检测试剂盒说明书紫外分光光度法注意：本产品试剂有所变动，请注意并严格按照该说明书操作。
货号：BC0530 规格：50T/48S产品组成：使用前请认真核对试剂体积与瓶内体积是否一致，有疑问请及时联系索莱宝工作人员。
试剂名称 规格 保存条件 提取液 液体60 mL×1瓶 2-8℃保存 试剂一 液体45 mL×1瓶 2-8℃保存 试剂二A 粉剂×1支 -20℃保存 试剂二B 粉剂×1支 -20℃保存 试剂二C 粉剂×2支 -20℃保存 试剂二D 粉剂×1支 -20℃保存 试剂三 液体45μL×1瓶 2-8℃保存 试剂四液体20μL×1瓶2-8℃保存溶液的配制：1、 试剂二A ：临用前加入1mL 蒸馏水，用不完的试剂-20℃分装保存4周，避免反复冻融；2、 试剂二B ：临用前加入1mL 蒸馏水，用不完的试剂-20℃分装保存4周，避免反复冻融；3、 试剂二C ：临用前取一支加入0.25mL 蒸馏水，用不完的试剂-20℃分装保存2周，避免反复冻融；4、 试剂二D ：临用前加入1mL 蒸馏水，用不完的试剂-20℃分装保存4周，避免反复冻融；5、 试剂三：临用前根据用量按照试剂三:蒸馏水为2μL :13μL （约3T ）的体积比例充分混匀，现用现配；6、 试剂四：临用前根据用量按照试剂四:蒸馏水为4μL :65μL （约13T ）的体积比例充分混匀，现用现配；7、 工作液的配制：根据样本量按试剂一：试剂二A ：试剂二B ：试剂二C ：试剂二D =22mL ：0.5mL ：0.5mL ：0.25mL ：0.5mL （约29T ）的比例配制工作液，现用现配；产品说明：磷酸果糖激酶（Phosphofructokinase ，PFK ，EC 2.7.1.11）广泛存在于动物、植物、微生物和培养细胞中，负责将果糖-6-磷酸和ATP 转化为果糖-1,6二磷酸和ADP ，是糖酵解过程的关键调节酶之一。
多烯磷脂酰胆碱注射液与果糖注射液的配伍稳定性研究
多烯磷脂酰胆碱注射液与果糖注射液的配伍稳定性研究发表时间：2015-11-18T14:24:04.250Z 来源：《中国耳鼻咽喉头颈外科》2015年9月第9期供稿作者：姜国伟，易清清，孟拥军*[导读] 上海市嘉定区中心医院上海配伍溶液会慢慢产生沉淀，因此配伍好的药液建议保存在4℃冰箱中，并在12h内滴完；25℃保存，2h内滴完。
【中图分类号】R942 【文献标识码】A姜国伟，易清清，孟拥军*上海市嘉定区中心医院上海 201800【摘要】目的：对多烯磷脂酰胆碱注射液与果糖注射液进行配伍探讨，为临床使用提供实验依据。
方法：将不同浓度的多烯磷脂酰胆碱注射液与果糖注射液进行配伍，在4℃及25℃保存下观察配伍溶液的外观，测定配伍溶液的pH值，以及配伍溶液中多烯磷脂酰胆碱的含量变化。
结果：在4℃时，12h内配伍溶液的外观、pH及溶液多烯磷脂酰胆碱的含量变化不大；而25℃时，2h内配伍溶液的外观、pH值及多烯磷脂酰胆碱的含量变化不大，超过2h，则有沉淀产生。
结论：多烯磷脂酰胆碱注射液与果糖注射液配伍，4℃保存时，12h内可用；25℃保存时，2h内可用。
【关键词】多烯磷脂酰胆碱注射液，果糖注射液，配伍Discussion of Polyene Phosphatidylcholine Injection and Fructose Injection CompatibilityJiang Guowei, Yi Qingqing, Meng Yongjun*Jiading Hospital, Shang Hai China 201800【Abstract】Objective: Making a discussion of Polyene Phosphatidylcholine injection and Fructose injection compatibility, to provide an experimental basis for clinical.Methods: Putting different concentrations of polyene phosphatidyl choline injection compatibility with fructose injection, and at 4℃and 25℃ observe the appearance of the compatibility solution, determination the compatibility pH, and the variation of polyene phosphatidyl choline injection. Results: At 4℃, the appearance of the compatibility solution, solution pH and polyene phosphatidylcholine changed little in 12h, while 25℃, the appearance of the compatibility of solution, pH and polyene phosphatidylcholine content changed little in 2h. Conclusion: The compatibility, of polyene phosphatidyl choline and fructose injection can been used within 4℃ in 12h, while saving within 25℃, 2h.【Keys】Polyene Phosphatidylcholine Injection, Fructose Injection, Compatibility果糖和葡萄糖同为糖源性能量物质，利于维持血糖水平，减少肝糖源分解以及节约蛋白质。
PEG修饰脂质体对阿霉素载药量的影响
・论著・PEG 修饰脂质体对阿霉素载药量的影响王向涛,李 沙,张小滨,侯新朴△(北京大学药学院药剂学系,北京 100083)[关键词]脂质体;阿霉素;药物载体;聚乙烯二醇类[摘 要]目的:在以阿霉素为模型药物用复乳法制备PEG 修饰脂质体实验中,发现当制备完毕立即上柱分离时,包封率约为50%;但当样品放置过夜后再测包封率却为98%。
本文通过实验分析该现象产生的原因。
方法:用不同方法制备PEG 修饰的空白脂质体,考察是否仍能吸附阿霉素;制备无PEG 修饰的空白脂质体,考察对阿霉素有无吸附作用;设计不同的PEG 2阿霉素配比,在不同的孵育时间取样测定对阿霉素的吸附量;将阿霉素溶液直接与PEG 20000混合孵育,考察无脂质体时PEG 对阿霉素的吸附;另选土霉素为模型药物,考察是否仍然存在吸附作用。
结果:不同方法制备的PEG 修饰的空白脂质体对阿霉素都有吸附;无PEG 修饰的空白脂质体对阿霉素无吸附作用;PEG 对阿霉素的吸附进程受PEG 2阿霉素配比、孵育时间的影响;单独的PEG 20000对阿霉素也能很好地吸附;以土霉素代替阿霉素,这种吸附作用仍然存在。
结论:该现象是由于PEG 链对体相中阿霉素或土霉素的吸附而引起的;这种吸附可能是阿霉素的羟基或氨基对PEG 链上氧原子之间的氢键作用。
提示这种相互作用的存在能够提高PEG 修饰的长循环脂质体,以及含PEG 材料的微囊、微球等对这类多羟基或多氨基小分子药物的载药量。
[中图分类号]R97911 [文献标识码]A [文章编号]16712167X (2002)0320286204The effect of PEG 2modif ied liposomes on adriamycin entrapmentWAN G Xiang 2Tao ,L I Sha ,ZHAN G Xiao 2Bin ,HOU Xin 2Pu △(Department of Pharmaceutics ,Peking University School of Pharmaceutical sciences ,Beijing 100083,China )KE Y WOR DS Liposomes ;Doxorubicin ;Drug carriers ;Polyethylene glycolsSUMMAR Y Objective :To analyse the reason for a strange phenomenon in adriamycin entrapment byliposomes prepared by double emulsion method.Methods :PEG 2modified blank liposomes were prepared by different methods ,and blank liposomes without PEG 2modification were also prepared in order to see whether the similar phenomenon would occur ;different PEG 2adriamycin ratios and different incu 2bation times were designed to observe their effect on absorption ;mixture composed of PEG 20000and adriamycin was dialysed to find the absorption between PEG itself and adriamycin.R esults :The phe 2nomenon was unaffected by preparation method and liposome itself ,but only affected by the presence of PEG chains around PEG 2modified liposomes ;both the PEG 2adriamycin ratio and incubation time af 2fected the absorption of adriamycin to PEG;the PEG alone could absorb adriamycin well ;the absorp 2tion occurred when oxytetracycline was in place of adriamycin.Conclusion :The reason for the phe 2nomenon was the absorption of adriamycin or oxytetracycline molecules to PEG chains where multiple hydrogen bonds might be formed.It is suggested PEG 2modified liposomes or PEG 2containing carriers such as microencapsules and microspheres would increase the entrapment efficiency or carrying amount of this kind of low molecular drugs which contain a number of hydroxies or aminos.(J Peki ng U niv [Health Sci ],2002,34:286-289) 复乳法用PEG 修饰脂质体包封阿霉素时,制备的脂质体如果立即过Sephadex G 250柱,则可明显基金项目:国家自然科学基金(30171114)资助Supported by National Sciences Foudation of China (30171114)△Corresponding author email ,houxinpu @・682・JOURNAL OF PEKIN G UNIV ERSITY (HEAL TH SCIENCES ) Vol.34 No.3 J un.2002看到阿霉素脂质体和游离阿霉素两条红色色带,测得包封率为50%。
氢化蛋黄卵磷脂精制工艺和吸附过程
氢化蛋黄卵磷脂精制工艺和吸附过程严喜鸾;李家伟;李奕潼;朱建航【摘要】大孔吸附树脂法精制氢化鲜鸭蛋黄卵磷脂和吸附过程研究.结果表明,从6种具有代表性的树脂中筛选出SP825大孔非极性树脂,高径比为14:1,上样浓度为3 mg·mL-1,上样流速为1.5 BV·h-1,95％乙醇溶液为洗脱液,所得产品纯度为91.13％,回收率为89.17％.等温吸附过程是熵增过程,拟合结果更符合Langmuir 方程.【期刊名称】《南昌大学学报（理科版）》【年(卷),期】2019(043)001【总页数】5页(P76-80)【关键词】大孔吸附树脂;卵磷脂;分离纯化;吸附【作者】严喜鸾;李家伟;李奕潼;朱建航【作者单位】南昌大学资源环境与化工学院;南昌大学资源环境与化工学院;南昌大学资源环境与化工学院;南昌大学资源环境与化工学院【正文语种】中文【中图分类】TQ460.6+1卵磷脂(Phosphatidylcholine，简称PC)，也称为磷脂酰胆碱，被誉为“第三营养素”[1-2]。
卵磷脂一般从大豆，蛋黄或是动物脑中提取，蛋黄中卵磷脂含量约10%，相比大豆和动物脑中含量更高，并且蛋黄卵磷脂具有良好的氧化稳定性。
工业生产中，卵磷脂的亲水性和乳化性限制了其应用范围。
黄国文[3]、赵俊延等人使用Pd/C催化剂与二氯甲烷混合进行大豆粉末磷脂的催化加氢实验。
章飞[4]、任其龙等人以大豆卵磷脂(SPC)为原料，5%Pd/C催化剂在高压釜中进行卵磷脂氢化反应，得到PC纯度为65%的氢化大豆卵磷脂(HSPC)，并通过柱层析法纯化得到高纯度大豆卵磷脂。
当前，对于氢化蛋黄卵磷脂的研究甚少，因此氢化蛋黄卵磷脂具有广泛的开发空间。
目前，国内外卵磷脂的纯化方法主要有溶剂萃取法、沉淀法、柱层析法、超临界萃取法等[5-8]。
大孔吸附树脂柱层析已广泛应用于物质的分离纯化[7-8]，具有环保、吸附能力强、可重复使用等优点。
卵磷脂由磷脂酰胆碱(PC)、磷脂酰乙醇胺(PE)、磷脂酰丝氨酸(PS)等组成[11]，甘油脂类物质为主要杂质。
大豆浓缩磷脂组成研究
中性油是伴随物质主要组成部分， 约占 30%~ 50%， 其含量主要受脱胶工艺影响。此外， 为改善磷脂流 动性， 也可适当添加一定比例中性油或脂肪酸。 糖脂是伴随物质重要组成部分， 约占 15%， 对于这 部分物质研究仍在进行， 其可能是影响浓缩磷脂透明 度重要因素， 是制备食品级浓缩磷脂需去除物质〔26〕 。 色素主要由大豆原料带入的类胡萝卜素、 叶绿 素、 叶黄素等及大豆在制油过程中生成褐色物或棕色 物或类黑素和卟啉类物质。大量褐色物在毛油水化 前已形成， 这些色泽形成很可能是毛油在汽提过程中 形成， 磷脂褐变反应是醛醇缩合反应， 以碱性物质作 为缩合催化剂， 而磷脂中磷脂酰胆碱正是这种缩合反 应催化剂； 且褐色物质一旦形成， 便很难脱除〔27〕 。其 它影响大豆磷脂色泽因素还有大豆产地、 大豆成熟程 度、 大豆加工预处理、 大豆轧胚厚度、 溶剂浸出温度、 水化脱胶条件、 磷脂加工条件等〔28〕 。 游离糖类物质占浓缩磷脂总质量 5% 左右， 这些
此外磷脂类物质还包括溶血磷脂酰胆碱lpc溶血磷脂酰丝氨酸lps溶血磷脂酰乙醇胺lpe胞苷二磷酸cdp酰化磷脂酰乙醇胺ape磷脂酰丙氨酸pala等微量磷脂类物质pc脂肪酸组成棕榈酸棕榈油酸硬脂酸亚油酸亚麻酸145502040410996464558大豆浓缩磷脂伴随物质组成在大豆浓缩磷脂中可将磷脂类物质外的物质统称为伴随物质主要包括中性油色素甾醇生育酚碳水化合物及水分无机固杂等微量组分正是其中一些物质及其衍生物影响浓缩磷脂透明度和色泽用不同方法将这些物质从磷脂中进一步去除可得到食品级磷脂和粉末磷脂
2009 年第 9 期
粮食与油脂
1
大 豆 浓 缩 磷 脂 组 成 研 究
2 刘方波 1， ， 刘元法 1， 王兴国 1 （1. 江南大学食品学院， 江苏无锡 214122； 2. 山东渤海油脂工业有限公司， 山东博兴 256500）
氧化铝柱层析分离磷脂工艺的洗脱研究
２００６年第３７卷第９期《浙江化工》文章编号：１００６－４１８４（２００６）０９－０００３－０２氧化铝柱层析分离磷脂工艺的洗脱研究＊柳叶，吕秀阳（浙江大学化学工程与生物工程学系，浙江杭州３１００２７）摘要：开展了不同洗脱液组成对氧化铝柱上磷脂洗脱曲线影响的实验研究。
分别用甲醇和甲醇－氨水体系做为洗脱剂进行洗脱，得到了最佳的洗脱液组成：甲醇：氨水＝２０：１（ｖ／ｖ）。
在此条件下，卵磷脂ＰＣ和脑磷脂ＰＥ的洗脱率分别达到了９８．６％和３２．３％。
这为用氧化铝柱层析来同时制备卵磷脂和脑磷脂的工艺的工业化研究奠定了重要基础。
关键词：氧化铝；洗脱；柱层析；卵磷脂；脑磷卵磷脂和脑磷脂是两种重要的磷脂，化学名称分别为磷脂酰胆碱（ｐｈｏｓｐｈｏｔｉｄｙｌｃｈｏｌｉｎｅ，简称ＰＣ）和磷脂酰乙醇胺（ＰｈｏｓｐｈａｔｉｄｙｌＥｔｈａｎｏｌａｍｉｎｅ，简称ＰＥ）。
高纯度磷脂产品具有无异味、乳化性能强等特点，应用更为广阔，国内外需求量在逐年增加。
目前纯度大于９５％的磷脂产品我国尚依赖进口，因此，开发出适合我国国情的高纯度磷脂制备方法具有重要的意义。
国内外对磷脂精制方法的研究较多［１￣５］，其中柱层析法由于分离效果好、工艺简单、投资少、处理量较大，因而成为磷脂精制方法的研究热点。
近年来本课题组致力于高纯度磷脂制备方法的研究［５，６］，本实验进一步对其洗脱特性进行了考察。
国内外未见相关的报道。
１实验部分１．１原料与试剂自制粗磷脂。
经高效液相色谱分析，其中ＰＣ含量为７９．６％，ＰＥ含量为１４．８％。
氧化铝（中性，颗粒尺寸０．０７４～０．１５ｍｍ，上海五四化学试剂有限公司）；在１２０℃下脱水活化１２ｈ，活化好的氧化铝转移至干燥器中，冷却后，加入８％去离子水。
密封存放７２ｈ以上，以备柱层析用。
此时氧化铝的含水量为８％（ｗｔ％）［６］。
ＨＰＬＣ用乙腈和甲醇（色谱纯，ＭＥＲＣＫ）；其余为分析纯。
１．２仪器和设备Ｒ－２０１旋转蒸发器（上海申生科技有限公司）；ＤＺＦ－６０２１真空干燥器（上海精宏实验设备有限公司）；色谱柱（１／２ｉｎ×２０ｃｍ，经水标定柱有效体积为２３ｍＬ，天津科器高新技术公司）；２Ｊ－Ｗ柱塞计量泵（杭州之江石化装备有限公司）；Ａｇｉｌｅｎｔ１１００型高效液相色谱仪（安捷伦科技有限公司）。