Darunavir_206361-99-1_CoA_MedChemExpress
高效液相色谱法测定糕点和果汁中纳他霉素的方法学验证
高效液相色谱法测定糕点和果汁中纳他霉素的方法学验证作者：卢薇周美丽来源：《食品安全导刊》2024年第03期摘要：为了验证宝鸡市食品药品检验检测中心是否有能力开展纳他霉素新方法《食品安全国家标准食品中纳他霉素的测定》（GB 5009.286—2022）的检测项目。
本文使用高效液相色谱仪，对糕点和果汁中纳他霉素进行分析，外标法定量，验证了本方法的线性关系、方法检出限、精密度和回收率。
结果表明，纳他霉素在0.5～10.0 μg·mL-1呈现良好的线性关系，方法检出限、精密度和回收率均能达到标准方法要求。
因此，本实验室具备检测纳他霉素的能力。
关键词：纳他霉素；外标法；高效液相色谱仪；方法学验证Methodological Validation of High Performance Liquid Chromatography for the Determination of Natamycin in Pastries and Fruit JuiceLU Wei， ZHOU Meili（Baoji Food and Drug Inspection and Testing Center， Baoji 721000， China）Abstract： In order to verify whether the Baoji Food and Drug Inspection and Testing Center has the ability to carry out the testing project of the new method GB 5009.286—2022 for natamycin. This article uses a high-performance liquid chromatography to analyze natamycin in pastries and fruit juice. The external standard method is used for quantification to verify the linear relationship， detection limit， precision， and recovery rate of this method. The results showed that natamycin exhibited a good linear relationship within the range from 0.5 μg·mL-1 to10 μg·mL-1， and the detection limit，precision， and recovery rate of the method can meet the requirements of the standard method. Therefore， our laboratory has the ability to detect natamycin.Keywords： natamycin; external standard method; high-performance liquid chromatography; methodology validation納他霉素，又名匹马霉素，是一种外观呈近白色或奶油黄色，无臭无味的结晶粉末，它是天然的多烯大环内酯类物质，对大部分的霉菌和酵母菌都具有极强的抑制作用[1]。
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NATURE NEUROSCIENCENAT REV CANCER NATURE REVIEWS CANCERNAT REV DRUG DISCOV NATURE REVIEWS DRUG DISCOVERYNAT REV IMMUNOL NATURE REVIEWS IMMUNOLOGYNAT REV NEUROSCI NATURE REVIEWS NEUROSCIENCENEUROLOGY NEUROLOGYNEURON NEURONNEUROPSYCHOPHARMACOL NEUROPSYCHOPHARMACOLOGYNEUROSCI BIOBEHAV R NEUROSCIENCE AND BIOBEHAVIORAL REVIEWSNEW ENGL J MED NEW ENGLAND JOURNAL OF MEDICINEOBES REV Obesity ReviewsONCOGENE ONCOGENEPHARMACOL REV PHARMACOLOGICAL REVIEWSPHARMACOL THERAPEUT PHARMACOLOGY & THERAPEUTICSPHYSIOL REV PHYSIOLOGICAL REVIEWSPHYSIOLOGY PHYSIOLOGYPLOS MED PLOS MEDICINEPROG NEUROBIOL PROGRESS IN NEUROBIOLOGYPROG RETIN EYE RES PROGRESS IN RETINAL AND EYE RESEARCHPSYCHOL BULL PSYCHOLOGICAL BULLETINPSYCHOL REV PSYCHOLOGICAL REVIEWREV MED VIROL REVIEWS IN MEDICAL VIROLOGYSCHIZOPHRENIA BULL SCHIZOPHRENIA BULLETINSEMIN CANCER BIOL SEMINARS IN CANCER BIOLOGYSEMIN IMMUNOL SEMINARS IN IMMUNOLOGYSTEM CELLS STEM CELLSSTROKE STROKETHORAX THORAXTRENDS COGN SCI TRENDS IN COGNITIVE SCIENCESTRENDS ENDOCRIN 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APPL PHYSIOL JOURNAL OF APPLIED PHYSIOLOGYJ CLIN ENDOCR METAB JOURNAL OF CLINICAL ENDOCRINOLOGY AND METABOLISM J CLIN MICROBIOL JOURNAL OF CLINICAL MICROBIOLOGYJ COMP NEUROL JOURNAL OF COMPARATIVE NEUROLOGYJ IMMUNOL JOURNAL OF IMMUNOLOGYJ INFECT DIS JOURNAL OF INFECTIOUS DISEASESJ MED CHEM JOURNAL OF MEDICINAL CHEMISTRYJ NEUROCHEM JOURNAL OF NEUROCHEMISTRYJ NEUROPHYSIOL JOURNAL OF NEUROPHYSIOLOGYJ NUTR JOURNAL OF NUTRITIONJ PHARMACOL EXP THER JOURNAL OF PHARMACOLOGY AND EXPERIMENTAL THERAPE J PHYSIOL-LONDON JOURNAL OF PHYSIOLOGY-LONDONJ UROLOGY JOURNAL OF UROLOGYJ VIROL JOURNAL OF VIROLOGYKIDNEY INT KIDNEY INTERNATIONALNEUROIMAGE NEUROIMAGENEUROSCIENCE NEUROSCIENCEPEDIATRICS PEDIATRICSRADIOLOGY RADIOLOGYTRANSPLANTATION TRANSPLANTATIONVIROLOGY VIROLOGYNATURE NATUREP NATL ACAD SCI USA PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES SCIENCE SCIENCEANN NY ACAD SCI ANNALS OF THE NEW YORK ACADEMY OF SCIENCESISSN大类名称复分大类分区是否top期刊2009年影响因子1680-7316地学1Y 4.881 0003-0007地学1Y 6.123 0930-7575地学1Y 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Diva Decloaker 10X Pretreatment Reagent 说明书
Intended Use:For In Vitro Diagnostic UseHeat induced antigen retrieval of formalin-fixed paraffin-embedded (FFPE) tissues for immunohistochemistry (IHC) procedures. The clinical interpretation of any staining or its absence should be complimented by morphological studies using proper controls and should be evaluated within the context of the patient's clinical history and other diagnostic tests by a qualified pathologist.Summary & Explanation:Diva Decloaker is a heat retrieval solution that is compatible with virtually all antibodies and eliminates the need for multiple buffers including citrate buffer, EDTA or high pH tris buffers. Antibody titers are doubled and tripled when compared to citrate buffer, pH 6.0. Diva Decloaker incorporates Assure™ tech nology, a color-coded high temperatures pH indicator solution. The end-user is assured by visual inspection that the solution is at the correct dilution and pH. This product is specially formulated for superior pH stability at high temperatures and will help prevent the possibility of losing pH sensitive antigens. Diva Decloaker is non-toxic, non-flammable, odorless and sodium azide and thimerosal free.Known Applications:Immunohistochemistry (formalin-fixed paraffin-embedded tissues) Supplied As:100mlDiva Decloaker, 10X concentrate (DV2004LX)500mlDiva Decloaker, 10X concentrate (DV2004MX)Materials and Reagents (Needed But Not Provided): Microscope slides, positively chargedDesert Chamber* (Drying oven)Positive and negative tissue controlsXylene (Could be substituted with xylene substitute*)Ethanol or reagent alcoholDecloaking Chamber* (Pressure cooker)Deionized or distilled waterWash buffer*(TBS/PBS)Enzyme digestion*Avidin-Biotin Blocking Kit*(Labeled Streptavidin Kits Only) Peroxidase block*Protein block*Primary antibody*Negative control reagents*Detection kits*Detection components*Chromogens*Hematoxylin*Bluing reagent*Mounting medium** Biocare Medical Products: Refer to a Biocare Medical catalog for further information regarding catalog number and ordering information. Certain reagents listed above are based on specific application and detection system used. Storage and Stability:Store at room temperature. Do not use after expiration date printed on vial. If reagents are stored under conditions other than those specified in the package insert, they must be verified by the user. Diluted reagents should be used promptly; any remaining reagent should be stored at room temperature.Protocol Recommendations:1. Deparaffinize tissues and hydrate to water. If necessary, block for endogenous peroxidase and wash in DI water.2. Dilute concentrated Diva Decloaker at a ratio of 1:10 (1 ml Diva to 9 ml of deionized water).3. Place slides into 1X retrieval solution in a slide container (e.g. Coplin Jar, Tissue -Tek™ staining dish or metal slide canister).4. Retrieve sections under pressure using Biocare's Decloaking Chamber. Follow the recommendations on the antibody data sheet and Decloaking Chamber User Manual.5. Check solution for appropriate color change. (See Technical Note #1)6. Gently rinse by gradually adding DI water to the solution, then remove slides and rinse with DI water.Technical Notes:1. Concentrated Diva Decloaker is a bright yellow color. RTU or 1X solution is a pale yellow color. When the solution reaches 80-125°C, the solution turns yellow and indicates that the high temperature solution is at correct pH. Should the pH rise above 7.0, the solution turns a fuschia red color. Should the pH drop too low, thesolution turns a pink color.2. If using Biocare’s Desert Chamber Pro (a programmable turbo-action drying oven), dry sections at 25ºC overnight or at 37ºC for 30-60 minutes and then dry slides at 60ºC for 30 minutes.3. Use positive char ged slides (use Biocare’s Kling-On HIER Slides) and cut tissues at 4-5 microns. Do not use any adhesives in the water bath. Poor fixation and processing of tissues will cause tissue sections to fall off the slides, especially fatty tissues such as breast. Tissues should be fixed a minimum of 6-12 hours.4. Protocol time and temperatures for HIER can vary depending on the Decloaking Chamber model used. Please refer to the relevant Decloaking Chamber manual for appropriate protocol times and temperatures.Limitations:The protocols for a specific application can vary. These include, but are not limited to: fixation, heat-retrieval method, incubation times, tissue section thickness and detection kit used. Due to the superior sensitivity of these unique reagents, the recommended incubation times and titers listed are not applicable to other detection systems, asresults may vary. The data sheet recommendations and protocols are based on exclusive use of Biocare products. Ultimately, it is the responsibility of the investigator to determine optimal conditions. The clinical interpretation of any positive or negative staining should be evaluated within the context of clinical presentation, morphology and other histopathological criteria by a qualified pathologist. The clinical interpretation of any positive or negative staining should be complemented by morphological studies using proper positive and negative internal and external controls as well as other diagnostic tests.Catalog Number: DV2004 LX, MX Description: 100, 500 ml, concentrateQuality Control:Refer to CLSI Quality Standards for Design and Implementation of Immunohistochemistry Assays; Approved Guideline-Second edition (I/LA28-A2). CLSI Wayne, PA, USA (). 2011 Precautions:1. This product is not classified as hazardous. The preservative used in this reagent is Proclin 300 and the concentration is less than 0.25%. Overexposure to Proclin 300 can cause skin and eye irritation and irritation to mucous membranes and upper respiratory tract. The concentration of Proclin 300 in this product does not meet the OSHA criteria for a hazardous substance. Wear disposable gloves when handling reagents.2. Specimens, before and after fixation, and all materials exposed to them should be handled as if capable of transmitting infection and disposed of with proper precautions. Never pipette reagents by mouth and avoid contacting the skin and mucous membranes with reagents and specimens. If reagents or specimens come in contact with sensitive areas, wash with copious amounts of water.3. Microbial contamination of reagents may result in an increase in nonspecific staining.4. Incubation times or temperatures other than those specified may give erroneous results. The user must validate any such change.5. Do not use reagent after the expiration date printed on the vial.6. The SDS is available upon request and is located at /.7. Consult OSHA, federal, state or local regulations for disposal of any toxic substances. Proclin is a trademark of Rohm and Haas Company, or of its subsidiaries or affiliates.Troubleshooting:Follow the antibody specific protocol recommendations according to data sheet provided. If atypical results occur, contact Biocare's Technical Support at 1-800-542-2002.。
生物质谱技术在腺相关病毒（AAV）载体制剂质量控制中的应用
学 报Journal of China Pharmaceutical University 2023，54（6）：682 - 694682生物质谱技术在腺相关病毒（AAV）载体制剂质量控制中的应用李孟效，李惠琳*（中山大学药学院，广州 510006）摘 要 腺相关病毒（adeno-associated virus，AAV）是一种基因治疗中常用的病毒载体。
由于其安全性较高且能够靶向多种细胞，在临床前和临床研究中得到了较多的应用。
不过在设计和生产的过程中，AAV载体有着诸多会影响其安全性和疗效的关键质量属性。
生物质谱技术的发展和应用为生物大分子的研究提供了一个便捷的平台，尤其是在蛋白质序列、结构和相互作用方面。
对于AAV载体而言，质谱技术可以实现衣壳蛋白比率、翻译后修饰、血清型、空衣壳比率的测定或表征，从而协助对AAV载体的质量控制。
与现有方法相比，质谱技术具有样品需求量少、分析快速灵敏、适用于完整AAV载体的分析和质量分辨率高的优点，并且可以区分空衣壳、满衣壳和部分包封的衣壳。
未来，通过将更加高效的蛋白质分离技术与质谱技术联用、开发新的信息处理软件平台和新的质谱检测方法，质谱技术有望在AAV载体的设计和生产中发挥更加重要的作用。
关键词腺相关病毒；质谱；质量控制；非变性质谱；电荷检测质谱中图分类号R392 文献标志码 A 文章编号1000 -5048（2023）06 -0682 -13doi：10.11665/j.issn.1000 -5048.2023062901引用本文李孟效，李惠琳.生物质谱技术在腺相关病毒（AAV）载体制剂质量控制中的应用［J］.中国药科大学学报，2023，54（6）：682–694.Cite this article as：LI Mengxiao，LI Huilin. Application of biological mass spectrometry in quality control of adeno-associated virus carrier preparations［J］.J China Pharm Univ，2023，54（6）：682–694.Application of biological mass spectrometry in quality control of adeno-asso⁃ciated virus carrier preparationsLI Mengxiao, LI Huilin*School of Pharmaceutical Science, Sun Yat-Sen University, Guangzhou 510006, ChinaAbstract Adeno-associated virus (AAV) is a common viral vector used in gene therapy.Because of its high safe⁃ty and its ability to target a variety of cells, it has been widely used in preclinical and clinical studies.However, during the design and production, AAV vectors have many key quality attributes that affect their safety and efficacy. The development and application of biological mass spectrometry technology provides a convenient platform for the research on biological macromolecules, especially in the aspects of protein sequence, structure and interac⁃tion.For AAV vectors, mass spectrometry can facilitate the determination or characterization of capsid protein ratio, post-translational modification, serotype, and empty capsid ratio, thus assisting in the quality control of AAV pared with the existing methods, mass spectrometry has the advantages of smaller amount of sample size, faster and more sensitive analysis, being more suitable for the analysis of complete AAV vectors with higher mass resolution, and can distinguish empty capsids, full capsids and partial capsids.In the future, mass spectrometry technology is expected to play a more important role in the design and production of AAV vectors through the coupling of more efficient protein separation technology with mass spectrometry, the development of new information processing software platforms and new mass spectrometry detection techniques.Key words adeno-associated virus (AAV); mass spectrometry (MS); quality control; native MS; charge detec⁃tion MS收稿日期2023-06-29 *通信作者Tel：138****4968E-mail：lihlin6@基金项目国家自然科学基金资助项目（No.81872836，No.91953102）；广东省自然科学基金资助项目（No.2019A1515011265）第 54 卷第 6 期李孟效，等：生物质谱技术在腺相关病毒（AAV ）载体制剂质量控制中的应用This study was supported by the National Natural Science Foundation of China (No.81872836, No.91953102) and the National Nat⁃ural Science Foundation of Guangdong Province (No.2019A1515011265)1 腺相关病毒(AAV)概述基因治疗是指为了治疗目的而修改操纵基因表达或改变活细胞的基因，基因治疗的方式可以划分基因补充和基因编辑，基因补充是指将遗传物质导入需要治疗的靶细胞，基因编辑是指对细胞已有的缺陷基因进行修改和调控，但无论哪种方式都需要依赖特定的递送载体才能完成[1]。
超声误诊无性细胞瘤一例
超声误诊无性细胞瘤一例发布时间：2021-09-24T02:21:37.511Z 来源：《医师在线》2021年23期作者：超声误诊无性细胞瘤一例[导读] 卵巢无性细胞瘤是卵巢恶性生殖细胞瘤贺宇凡，曾思惠通讯作者（广东省妇幼保健院广东广州 511400）【摘要】：卵巢无性细胞瘤是卵巢恶性生殖细胞瘤，来源于尚未分化以前的原始生殖细胞，临床发病率低且缺乏典型早期症状，主要表现为腹痛及腹部肿块。
本文分析总结一例以急腹症就诊行急诊阴超检查的误诊病例报道，以期为无性细胞瘤的超声正确诊断提供经验。
【关键词】无性细胞瘤；卵巢1.临床资料患者，女，24岁，未婚，性生活正常。
因“停经41天，下腹痛4天，加重1天”来我院就诊。
现病史：患者平素月经规则，周期28-30天，经期3天。
2020-6-9无明显诱因出现下腹胀痛，间断发作，疼痛轻微，未予重视。
2020-6-11下腹胀痛较前加重，体位改变时明显，休息后好转，不伴发热、畏寒、寒战、恶心、呕吐等不适，外院就诊HGB：120g/l，腹部超声、泌尿系超声及阑尾区超声均未见明显异常，妇科超声示子宫前方低回声不均质肿块，建议上级医院治疗，现患者为求进一步诊治，我院就诊。
体格检查：体温：36.4℃，脉搏：120次/分，呼吸：20次/分，血压：116/79mmHg。
腹平软，无肌紧张，肠鸣音正常，下腹部轻压痛，无反跳痛。
妇检：阴道少许淡红色分泌物，宫颈举痛（+）及摇摆痛（+），后穹隆穿刺抽出不凝血0.5ml。
子宫前方巨大包块，活动度差，边界触及不清，子宫大小触及不清，有压痛。
实验室检查：尿HCG（+）。
婚育史：未婚，性生活正常。
G0P0.既往史：既往体健，既往检查：7个月前外院体检行妇科超声检查，未见异常。
2020-6-13经阴道超声检查所见：左卵巢大小38mm×23mm，右卵巢大小42mm×34mm，子宫上方见混合性包块，大小132mm×79mm，边界清，似与双卵巢分界清，CDFI:混合性包块边缘及内部见少许彩色血流信号。
稳定性英文版
HPLC ASSAY with DETERMINATION OF META-FLUOXETINE HCl.ANALYTICAL METHOD VALIDATION10 and 20mg Fluoxetine Capsules HPLC DeterminationFLUOXETINE HClC17H18F3NO•HClM.W. = 345.79CAS — 59333-67-4STABILITY INDICATINGA S S A Y V A L I D A T I O NMethod is suitable for:ýIn-process controlþProduct ReleaseþStability indicating analysis (Suitability - US/EU Product) CAUTIONFLUOXETINE HYDROCHLORIDE IS A HAZARDOUS CHEMICAL AND SHOULD BE HANDLED ONLY UNDER CONDITIONS SUITABLE FOR HAZARDOUS WORK.IT IS HIGHLY PRESSURE SENSITIVE AND ADEQUATE PRECAUTIONS SHOULD BE TAKEN TO AVOID ANY MECHANICAL FORCE (SUCH AS GRINDING, CRUSHING, ETC.) ON THE POWDER.ED. N0: 04Effective Date:APPROVED::HPLC ASSAY with DETERMINATION OF META-FLUOXETINE HCl.ANALYTICAL METHOD VALIDATION10 and 20mg Fluoxetine Capsules HPLC DeterminationTABLE OF CONTENTS INTRODUCTION........................................................................................................................ PRECISION............................................................................................................................... System Repeatability ................................................................................................................ Method Repeatability................................................................................................................. Intermediate Precision .............................................................................................................. LINEARITY................................................................................................................................ RANGE...................................................................................................................................... ACCURACY............................................................................................................................... Accuracy of Standard Injections................................................................................................ Accuracy of the Drug Product.................................................................................................... VALIDATION OF FLUOXETINE HCl AT LOW CONCENTRATION........................................... Linearity at Low Concentrations................................................................................................. Accuracy of Fluoxetine HCl at Low Concentration..................................................................... System Repeatability................................................................................................................. Quantitation Limit....................................................................................................................... Detection Limit........................................................................................................................... VALIDATION FOR META-FLUOXETINE HCl (POSSIBLE IMPURITIES).................................. Meta-Fluoxetine HCl linearity at 0.05% - 1.0%........................................................................... Detection Limit for Fluoxetine HCl.............................................................................................. Quantitation Limit for Meta Fluoxetine HCl................................................................................ Accuracy for Meta-Fluoxetine HCl ............................................................................................ Method Repeatability for Meta-Fluoxetine HCl........................................................................... Intermediate Precision for Meta-Fluoxetine HCl......................................................................... SPECIFICITY - STABILITY INDICATING EVALUATION OF THE METHOD............................. FORCED DEGRADATION OF FINISHED PRODUCT AND STANDARD..................................1. Unstressed analysis...............................................................................................................2. Acid Hydrolysis stressed analysis..........................................................................................3. Base hydrolysis stressed analysis.........................................................................................4. Oxidation stressed analysis...................................................................................................5. Sunlight stressed analysis.....................................................................................................6. Heat of solution stressed analysis.........................................................................................7. Heat of powder stressed analysis.......................................................................................... System Suitability stressed analysis.......................................................................................... Placebo...................................................................................................................................... STABILITY OF STANDARD AND SAMPLE SOLUTIONS......................................................... Standard Solution...................................................................................................................... Sample Solutions....................................................................................................................... ROBUSTNESS.......................................................................................................................... Extraction................................................................................................................................... Factorial Design......................................................................................................................... CONCLUSION...........................................................................................................................ED. N0: 04Effective Date:APPROVED::HPLC ASSAY with DETERMINATION OF META-FLUOXETINE HCl.ANALYTICAL METHOD VALIDATION10 and 20mg Fluoxetine Capsules HPLC DeterminationBACKGROUNDTherapeutically, Fluoxetine hydrochloride is a classified as a selective serotonin-reuptake inhibitor. Effectively used for the treatment of various depressions. Fluoxetine hydrochloride has been shown to have comparable efficacy to tricyclic antidepressants but with fewer anticholinergic side effects. The patent expiry becomes effective in 2001 (US). INTRODUCTIONFluoxetine capsules were prepared in two dosage strengths: 10mg and 20mg dosage strengths with the same capsule weight. The formulas are essentially similar and geometrically equivalent with the same ingredients and proportions. Minor changes in non-active proportions account for the change in active ingredient amounts from the 10 and 20 mg strength.The following validation, for the method SI-IAG-206-02 , includes assay and determination of Meta-Fluoxetine by HPLC, is based on the analytical method validation SI-IAG-209-06. Currently the method is the in-house method performed for Stability Studies. The Validation was performed on the 20mg dosage samples, IAG-21-001 and IAG-21-002.In the forced degradation studies, the two placebo samples were also used. PRECISIONSYSTEM REPEATABILITYFive replicate injections of the standard solution at the concentration of 0.4242mg/mL as described in method SI-IAG-206-02 were made and the relative standard deviation (RSD) of the peak areas was calculated.SAMPLE PEAK AREA#15390#25406#35405#45405#55406Average5402.7SD 6.1% RSD0.1ED. N0: 04Effective Date:APPROVED::HPLC ASSAY with DETERMINATION OF META-FLUOXETINE HCl.ANALYTICAL METHOD VALIDATION10 and 20mg Fluoxetine Capsules HPLC DeterminationED. N0: 04Effective Date:APPROVED::PRECISION - Method RepeatabilityThe full HPLC method as described in SI-IAG-206-02 was carried-out on the finished product IAG-21-001 for the 20mg dosage form. The method repeated six times and the relative standard deviation (RSD) was calculated.SAMPLENumber%ASSAYof labeled amountI 96.9II 97.8III 98.2IV 97.4V 97.7VI 98.5(%) Average97.7SD 0.6(%) RSD0.6PRECISION - Intermediate PrecisionThe full method as described in SI-IAG-206-02 was carried-out on the finished product IAG-21-001 for the 20mg dosage form. The method was repeated six times by a second analyst on a different day using a different HPLC instrument. The average assay and the relative standard deviation (RSD) were calculated.SAMPLENumber% ASSAYof labeled amountI 98.3II 96.3III 94.6IV 96.3V 97.8VI 93.3Average (%)96.1SD 2.0RSD (%)2.1The difference between the average results of method repeatability and the intermediate precision is 1.7%.HPLC ASSAY with DETERMINATION OF META-FLUOXETINE HCl.ANALYTICAL METHOD VALIDATION10 and 20mg Fluoxetine Capsules HPLC DeterminationLINEARITYStandard solutions were prepared at 50% to 200% of the nominal concentration required by the assay procedure. Linear regression analysis demonstrated acceptability of the method for quantitative analysis over the concentration range required. Y-Intercept was found to be insignificant.RANGEDifferent concentrations of the sample (IAG-21-001) for the 20mg dosage form were prepared, covering between 50% - 200% of the nominal weight of the sample.Conc. (%)Conc. (mg/mL)Peak Area% Assayof labeled amount500.20116235096.7700.27935334099.21000.39734463296.61500.64480757797.52000.79448939497.9(%) Average97.6SD 1.0(%) RSD 1.0ED. N0: 04Effective Date:APPROVED::HPLC ASSAY with DETERMINATION OF META-FLUOXETINE HCl.ANALYTICAL METHOD VALIDATION10 and 20mg Fluoxetine Capsules HPLC DeterminationED. N0: 04Effective Date:APPROVED::RANGE (cont.)The results demonstrate linearity as well over the specified range.Correlation coefficient (RSQ)0.99981 Slope11808.3Y -Interceptresponse at 100%* 100 (%) 0.3%ACCURACYACCURACY OF STANDARD INJECTIONSFive (5) replicate injections of the working standard solution at concentration of 0.4242mg/mL, as described in method SI-IAG-206-02 were made.INJECTIONNO.PEAK AREA%ACCURACYI 539299.7II 540599.9III 540499.9IV 5406100.0V 5407100.0Average 5402.899.9%SD 6.10.1RSD, (%)0.10.1The percent deviation from the true value wasdetermined from the linear regression lineHPLC ASSAY with DETERMINATION OF META-FLUOXETINE HCl.ANALYTICAL METHOD VALIDATION10 and 20mg Fluoxetine Capsules HPLC DeterminationED. N0: 04Effective Date:APPROVED::ACCURACY OF THE DRUG PRODUCTAdmixtures of non-actives (placebo, batch IAG-21-001 ) with Fluoxetine HCl were prepared at the same proportion as in a capsule (70%-180% of the nominal concentration).Three preparations were made for each concentration and the recovery was calculated.Conc.(%)Placebo Wt.(mg)Fluoxetine HCl Wt.(mg)Peak Area%Accuracy Average (%)70%7079.477.843465102.27079.687.873427100.77079.618.013465100.0101.0100%10079.6211.25476397.910080.8011.42491799.610079.6011.42485498.398.6130%13079.7214.90640599.413080.3114.75632899.213081.3314.766402100.399.618079.9920.10863699.318079.3820.45879499.418080.0820.32874899.599.4Placebo, Batch Lot IAG-21-001HPLC ASSAY with DETERMINATION OF META-FLUOXETINE HCl.ANALYTICAL METHOD VALIDATION10 and 20mg Fluoxetine Capsules HPLC DeterminationED. N0: 04Effective Date:APPROVED::VALIDATION OF FLUOXETINE HClAT LOW CONCENTRATIONLINEARITY AT LOW CONCENTRATIONSStandard solution of Fluoxetine were prepared at approximately 0.02%-1.0% of the working concentration required by the method SI-IAG-206-02. Linear regression analysis demonstrated acceptability of the method for quantitative analysis over this range.ACCURACY OF FLUOXETINE HCl AT LOW CONCENTRATIONThe peak areas of the standard solution at the working concentration were measured and the percent deviation from the true value, as determined from the linear regression was calculated.SAMPLECONC.µg/100mLAREA FOUND%ACCURACYI 470.56258499.7II 470.56359098.1III 470.561585101.3IV 470.561940100.7V 470.56252599.8VI 470.56271599.5(%) AverageSlope = 132.7395299.9SD Y-Intercept = -65.872371.1(%) RSD1.1HPLC ASSAY with DETERMINATION OF META-FLUOXETINE HCl.ANALYTICAL METHOD VALIDATION10 and 20mg Fluoxetine Capsules HPLC DeterminationSystem RepeatabilitySix replicate injections of standard solution at 0.02% and 0.05% of working concentration as described in method SI-IAG-206-02 were made and the relative standard deviation was calculated.SAMPLE FLUOXETINE HCl AREA0.02%0.05%I10173623II11503731III10103475IV10623390V10393315VI10953235Average10623462RSD, (%) 5.0 5.4Quantitation Limit - QLThe quantitation limit ( QL) was established by determining the minimum level at which the analyte was quantified. The quantitation limit for Fluoxetine HCl is 0.02% of the working standard concentration with resulting RSD (for six injections) of 5.0%. Detection Limit - DLThe detection limit (DL) was established by determining the minimum level at which the analyte was reliably detected. The detection limit of Fluoxetine HCl is about 0.01% of the working standard concentration.ED. N0: 04Effective Date:APPROVED::HPLC ASSAY with DETERMINATION OF META-FLUOXETINE HCl.ANALYTICAL METHOD VALIDATION10 and 20mg Fluoxetine Capsules HPLC DeterminationED. N0: 04Effective Date:APPROVED::VALIDATION FOR META-FLUOXETINE HCl(EVALUATING POSSIBLE IMPURITIES)Meta-Fluoxetine HCl linearity at 0.05% - 1.0%Relative Response Factor (F)Relative response factor for Meta-Fluoxetine HCl was determined as slope of Fluoxetine HCl divided by the slope of Meta-Fluoxetine HCl from the linearity graphs (analysed at the same time).F =132.7395274.859534= 1.8Detection Limit (DL) for Fluoxetine HClThe detection limit (DL) was established by determining the minimum level at which the analyte was reliably detected.Detection limit for Meta Fluoxetine HCl is about 0.02%.Quantitation Limit (QL) for Meta-Fluoxetine HClThe QL is determined by the analysis of samples with known concentration of Meta-Fluoxetine HCl and by establishing the minimum level at which the Meta-Fluoxetine HCl can be quantified with acceptable accuracy and precision.Six individual preparations of standard and placebo spiked with Meta-Fluoxetine HCl solution to give solution with 0.05% of Meta Fluoxetine HCl, were injected into the HPLC and the recovery was calculated.HPLC ASSAY with DETERMINATION OF META-FLUOXETINE HCl.ANALYTICAL METHOD VALIDATION10 and 20mg Fluoxetine Capsules HPLC DeterminationED. N0: 04Effective Date:APPROVED::META-FLUOXETINE HCl[RECOVERY IN SPIKED SAMPLES].Approx.Conc.(%)Known Conc.(µg/100ml)Area in SpikedSampleFound Conc.(µg/100mL)Recovery (%)0.0521.783326125.735118.10.0521.783326825.821118.50.0521.783292021.55799.00.0521.783324125.490117.00.0521.783287220.96996.30.0521.783328526.030119.5(%) AVERAGE111.4SD The recovery result of 6 samples is between 80%-120%.10.7(%) RSDQL for Meta Fluoxetine HCl is 0.05%.9.6Accuracy for Meta Fluoxetine HClDetermination of Accuracy for Meta-Fluoxetine HCl impurity was assessed using triplicate samples (of the drug product) spiked with known quantities of Meta Fluoxetine HCl impurity at three concentrations levels (namely 80%, 100% and 120% of the specified limit - 0.05%).The results are within specifications:For 0.4% and 0.5% recovery of 85% -115%For 0.6% recovery of 90%-110%HPLC ASSAY with DETERMINATION OF META-FLUOXETINE HCl.ANALYTICAL METHOD VALIDATION10 and 20mg Fluoxetine Capsules HPLC DeterminationED. N0: 04Effective Date:APPROVED::META-FLUOXETINE HCl[RECOVERY IN SPIKED SAMPLES]Approx.Conc.(%)Known Conc.(µg/100mL)Area in spikedSample Found Conc.(µg/100mL)Recovery (%)[0.4%]0.4174.2614283182.66104.820.4174.2614606187.11107.370.4174.2614351183.59105.36[0.5%]0.5217.8317344224.85103.220.5217.8316713216.1599.230.5217.8317341224.81103.20[0.6%]0.6261.3918367238.9591.420.6261.3920606269.81103.220.6261.3920237264.73101.28RECOVERY DATA DETERMINED IN SPIKED SAMPLESHPLC ASSAY with DETERMINATION OF META-FLUOXETINE HCl.ANALYTICAL METHOD VALIDATION10 and 20mg Fluoxetine Capsules HPLC DeterminationED. N0: 04Effective Date:APPROVED::REPEATABILITYMethod Repeatability - Meta Fluoxetine HClThe full method (as described in SI-IAG-206-02) was carried out on the finished drug product representing lot number IAG-21-001-(1). The HPLC method repeated serially, six times and the relative standard deviation (RSD) was calculated.IAG-21-001 20mg CAPSULES - FLUOXETINESample% Meta Fluoxetine % Meta-Fluoxetine 1 in Spiked Solution10.0260.09520.0270.08630.0320.07740.0300.07450.0240.09060.0280.063AVERAGE (%)0.0280.081SD 0.0030.012RSD, (%)10.314.51NOTE :All results are less than QL (0.05%) therefore spiked samples with 0.05% Meta Fluoxetine HCl were injected.HPLC ASSAY with DETERMINATION OF META-FLUOXETINE HCl.ANALYTICAL METHOD VALIDATION10 and 20mg Fluoxetine Capsules HPLC DeterminationED. N0: 04Effective Date:APPROVED::Intermediate Precision - Meta-Fluoxetine HClThe full method as described in SI-IAG-206-02 was applied on the finished product IAG-21-001-(1) .It was repeated six times, with a different analyst on a different day using a different HPLC instrument.The difference between the average results obtained by the method repeatability and the intermediate precision was less than 30.0%, (11.4% for Meta-Fluoxetine HCl as is and 28.5% for spiked solution).IAG-21-001 20mg - CAPSULES FLUOXETINESample N o:Percentage Meta-fluoxetine% Meta-fluoxetine 1 in spiked solution10.0260.06920.0270.05730.0120.06140.0210.05850.0360.05560.0270.079(%) AVERAGE0.0250.063SD 0.0080.009(%) RSD31.514.51NOTE:All results obtained were well below the QL (0.05%) thus spiked samples slightly greater than 0.05% Meta-Fluoxetine HCl were injected. The RSD at the QL of the spiked solution was 14.5%HPLC ASSAY with DETERMINATION OF META-FLUOXETINE HCl.ANALYTICAL METHOD VALIDATION10 and 20mg Fluoxetine Capsules HPLC DeterminationSPECIFICITY - STABILITY INDICATING EVALUATIONDemonstration of the Stability Indicating parameters of the HPLC assay method [SI-IAG-206-02] for Fluoxetine 10 & 20mg capsules, a suitable photo-diode array detector was incorporated utilizing a commercial chromatography software managing system2, and applied to analyze a range of stressed samples of the finished drug product.GLOSSARY of PEAK PURITY RESULT NOTATION (as reported2):Purity Angle-is a measure of spectral non-homogeneity across a peak, i.e. the weighed average of all spectral contrast angles calculated by comparing all spectra in the integrated peak against the peak apex spectrum.Purity Threshold-is the sum of noise angle3 and solvent angle4. It is the limit of detection of shape differences between two spectra.Match Angle-is a comparison of the spectrum at the peak apex against a library spectrum.Match Threshold-is the sum of the match noise angle3 and match solvent angle4.3Noise Angle-is a measure of spectral non-homogeneity caused by system noise.4Solvent Angle-is a measure of spectral non-homogeneity caused by solvent composition.OVERVIEWT he assay of the main peak in each stressed solution is calculated according to the assay method SI-IAG-206-02, against the Standard Solution, injected on the same day.I f the Purity Angle is smaller than the Purity Threshold and the Match Angle is smaller than the Match Threshold, no significant differences between spectra can be detected. As a result no spectroscopic evidence for co-elution is evident and the peak is considered to be pure.T he stressed condition study indicated that the Fluoxetine peak is free from any appreciable degradation interference under the stressed conditions tested. Observed degradation products peaks were well separated from the main peak.1® PDA-996 Waters™ ; 2[Millennium 2010]ED. N0: 04Effective Date:APPROVED::HPLC ASSAY with DETERMINATION OF META-FLUOXETINE HCl.ANALYTICAL METHOD VALIDATION10 and 20mg Fluoxetine Capsules HPLC DeterminationFORCED DEGRADATION OF FINISHED PRODUCT & STANDARD 1.UNSTRESSED SAMPLE1.1.Sample IAG-21-001 (2) (20mg/capsule) was prepared as stated in SI-IAG-206-02 and injected into the HPLC system. The calculated assay is 98.5%.SAMPLE - UNSTRESSEDFluoxetine:Purity Angle:0.075Match Angle:0.407Purity Threshold:0.142Match Threshold:0.4251.2.Standard solution was prepared as stated in method SI-IAG-206-02 and injected into the HPLC system. The calculated assay is 100.0%.Fluoxetine:Purity Angle:0.078Match Angle:0.379Purity Threshold:0.146Match Threshold:0.4272.ACID HYDROLYSIS2.1.Sample solution of IAG-21-001 (2) (20mg/capsule) was prepared as in method SI-IAG-206-02 : An amount equivalent to 20mg Fluoxetine was weighed into a 50mL volumetric flask. 20mL Diluent was added and the solution sonicated for 10 minutes. 1mL of conc. HCl was added to this solution The solution was allowed to stand for 18 hours, then adjusted to about pH = 5.5 with NaOH 10N, made up to volume with Diluent and injected into the HPLC system after filtration.Fluoxetine peak intensity did NOT decrease. Assay result obtained - 98.8%.SAMPLE- ACID HYDROLYSISFluoxetine peak:Purity Angle:0.055Match Angle:0.143Purity Threshold:0.096Match Threshold:0.3712.2.Standard solution was prepared as in method SI-IAG-206-02 : about 22mg Fluoxetine HCl were weighed into a 50mL volumetric flask. 20mL Diluent were added. 2mL of conc. HCl were added to this solution. The solution was allowed to stand for 18 hours, then adjusted to about pH = 5.5 with NaOH 10N, made up to volume with Diluent and injected into the HPLC system.Fluoxetine peak intensity did NOT decrease. Assay result obtained - 97.2%.ED. N0: 04Effective Date:APPROVED::HPLC ASSAY with DETERMINATION OF META-FLUOXETINE HCl.ANALYTICAL METHOD VALIDATION10 and 20mg Fluoxetine Capsules HPLC DeterminationSTANDARD - ACID HYDROLYSISFluoxetine peak:Purity Angle:0.060Match Angle:0.060Purity Threshold:0.099Match Threshold:0.3713.BASE HYDROLYSIS3.1.Sample solution of IAG-21-001 (2) (20mg/capsule) was prepared as per method SI-IAG-206-02 : An amount equivalent to 20mg Fluoxetine was weight into a 50mL volumetric flask. 20mL Diluent was added and the solution sonicated for 10 minutes. 1mL of 5N NaOH was added to this solution. The solution was allowed to stand for 18 hours, then adjusted to about pH = 5.5 with 5N HCl, made up to volume with Diluent and injected into the HPLC system.Fluoxetine peak intensity did NOT decrease. Assay result obtained - 99.3%.SAMPLE - BASE HYDROLYSISFluoxetine peak:Purity Angle:0.063Match Angle:0.065Purity Threshold:0.099Match Threshold:0.3623.2.Standard stock solution was prepared as per method SI-IAG-206-02 : About 22mg Fluoxetine HCl was weighed into a 50mL volumetric flask. 20mL Diluent was added. 2mL of 5N NaOH was added to this solution. The solution was allowed to stand for 18 hours, then adjusted to about pH=5.5 with 5N HCl, made up to volume with Diluent and injected into the HPLC system.Fluoxetine peak intensity did NOT decrease - 99.5%.STANDARD - BASE HYDROLYSISFluoxetine peak:Purity Angle:0.081Match Angle:0.096Purity Threshold:0.103Match Threshold:0.3634.OXIDATION4.1.Sample solution of IAG-21-001 (2) (20mg/capsule) was prepared as per method SI-IAG-206-02. An equivalent to 20mg Fluoxetine was weighed into a 50mL volumetric flask. 20mL Diluent added and the solution sonicated for 10 minutes.1.0mL of 30% H2O2 was added to the solution and allowed to stand for 5 hours, then made up to volume with Diluent, filtered and injected into HPLC system.Fluoxetine peak intensity decreased to 95.2%.ED. N0: 04Effective Date:APPROVED::HPLC ASSAY with DETERMINATION OF META-FLUOXETINE HCl.ANALYTICAL METHOD VALIDATION10 and 20mg Fluoxetine Capsules HPLC DeterminationSAMPLE - OXIDATIONFluoxetine peak:Purity Angle:0.090Match Angle:0.400Purity Threshold:0.154Match Threshold:0.4294.2.Standard solution was prepared as in method SI-IAG-206-02 : about 22mg Fluoxetine HCl were weighed into a 50mL volumetric flask and 25mL Diluent were added. 2mL of 30% H2O2 were added to this solution which was standing for 5 hours, made up to volume with Diluent and injected into the HPLC system.Fluoxetine peak intensity decreased to 95.8%.STANDARD - OXIDATIONFluoxetine peak:Purity Angle:0.083Match Angle:0.416Purity Threshold:0.153Match Threshold:0.4295.SUNLIGHT5.1.Sample solution of IAG-21-001 (2) (20mg/capsule) was prepared as in method SI-IAG-206-02 . The solution was exposed to 500w/hr. cell sunlight for 1hour. The BST was set to 35°C and the ACT was 45°C. The vials were placed in a horizontal position (4mm vials, National + Septum were used). A Dark control solution was tested. A 2%w/v quinine solution was used as the reference absorbance solution.Fluoxetine peak decreased to 91.2% and the dark control solution showed assay of 97.0%. The difference in the absorbance in the quinine solution is 0.4227AU.Additional peak was observed at RRT of 1.5 (2.7%).The total percent of Fluoxetine peak with the degradation peak is about 93.9%.SAMPLE - SUNLIGHTFluoxetine peak:Purity Angle:0.093Match Angle:0.583Purity Threshold:0.148Match Threshold:0.825 ED. N0: 04Effective Date:APPROVED::HPLC ASSAY with DETERMINATION OF META-FLUOXETINE HCl.ANALYTICAL METHOD VALIDATION10 and 20mg Fluoxetine Capsules HPLC DeterminationSUNLIGHT (Cont.)5.2.Working standard solution was prepared as in method SI-IAG-206-02 . The solution was exposed to 500w/hr. cell sunlight for 1.5 hour. The BST was set to 35°C and the ACT was 42°C. The vials were placed in a horizontal position (4mm vials, National + Septum were used). A Dark control solution was tested. A 2%w/v quinine solution was used as the reference absorbance solution.Fluoxetine peak was decreased to 95.2% and the dark control solution showed assay of 99.5%.The difference in the absorbance in the quinine solution is 0.4227AU.Additional peak were observed at RRT of 1.5 (2.3).The total percent of Fluoxetine peak with the degradation peak is about 97.5%. STANDARD - SUNLIGHTFluoxetine peak:Purity Angle:0.067Match Angle:0.389Purity Threshold:0.134Match Threshold:0.8196.HEAT OF SOLUTION6.1.Sample solution of IAG-21-001-(2) (20 mg/capsule) was prepared as in method SI-IAG-206-02 . Equivalent to 20mg Fluoxetine was weighed into a 50mL volumetric flask. 20mL Diluent was added and the solution was sonicated for 10 minutes and made up to volume with Diluent. 4mL solution was transferred into a suitable crucible, heated at 105°C in an oven for 2 hours. The sample was cooled to ambient temperature, filtered and injected into the HPLC system.Fluoxetine peak was decreased to 93.3%.SAMPLE - HEAT OF SOLUTION [105o C]Fluoxetine peak:Purity Angle:0.062Match Angle:0.460Purity Threshold:0.131Match Threshold:0.8186.2.Standard Working Solution (WS) was prepared under method SI-IAG-206-02 . 4mL of the working solution was transferred into a suitable crucible, placed in an oven at 105°C for 2 hours, cooled to ambient temperature and injected into the HPLC system.Fluoxetine peak intensity did not decrease - 100.5%.ED. N0: 04Effective Date:APPROVED::。
《临床肝胆病杂志》推荐使用的规范医学名词术语
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SCI_Chem 影响因子
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Organomet Chem0268-26052866 2.011 1.9220.272 267Electrochem Solid St1099-00628883 2.01 2.0260.596 268J Chem Eng Data0021-956815169 2.004 2.1150.295 269Comb Chem High T Scr1386-207314532 1.9750.268 269J Organomet Chem0022-328X217932 1.9920.527 271Thermochim Acta0040-603111408 1.989 2.0460.327 272J Mol Model1610-29403157 1.984 2.3010.378 273J Therm Anal Calorim1388-61509934 1.982 1.7420.243 274Int J Fatigue0142-11235248 1.976 1.9740.352 275J Vib Control1077-54631649 1.966 1.7360.672 276Chem Phys0301-010412935 1.957 2.0590.592 277J Mater Process Tech0924-013618426 1.953 2.1760.332 277Sensors-Basel1424-82207082 1.953 2.3950.321 279Biomed Chromatogr0269-38792861 1.945 1.8150.385 280J Fluorine Chem0022-11394998 1.939 1.9490.465ArticlesCited Half-life Eigenfactor ®Metrics ScoreArticleInfluenc e® Score8699.6 1.5750820.8448329.7 1.3598717.71238018 1.55663 4.8921768.20.2266114.294141 5.20.2278819.481121 3.70.1543615.607390 3.50.178418.8542370.015018.643126 2.40.054018.927207 6.50.108177.90837 3.20.01489 5.62318>10.00.013647.55389.40.008828.821867 5.10.27819 4.24368.40.00872 6.542227 5.50.53637 3.497299.90.01888.1211078 4.40.37491 5.1891191 2.40.20333 4.012473 1.80.05534 3.461368.20.03818 3.07130997.70.83183 2.99465 5.90.01757 3.141169 1.40.00944 3.344569 4.20.12336 3.006377.20.00647 2.73229 1.90.00155 3.27323 3.90.00716 4.441557.70.01045 2.068458 1.40.02121 2.833576 6.70.15215 2.032167.90.00342 3.148457 3.60.07856 2.503112 4.60.01202 1.97501 6.90.05008 1.9062220.00197 2.73916>10.00.003282.792Eigenfactor®Metrics286 2.60.0208 2.0046>10.00.00142 1.13896 2.80.01517 2.301119.20.00397 3.974>10.00.00086 1.894 44940.03848 1.5233>10.00.00076 2.537437.60.01013 2.301 63220.04884 2.393 2150.003483173 4.80.28954 1.5629>10.00.00211 3.009 138 5.80.01695 1.794 1015 1.70.02966 1.597 508.20.01226 2.37 160850.18182 1.40244 5.40.01448 3.213188.30.00431 2.295 107.10.00156 2.34 674 6.40.06437 1.6 1916 4.10.1766 1.469 480 5.40.05059 1.419 284>10.00.04014 1.60356 1.20.00194 1.703 617 3.70.05475 1.603 14797.80.17236 1.533 8907.80.09182 1.35 404 4.40.04405 1.352 239.20.00403 2.705 448 4.10.05929 1.252 507 3.50.04948 1.865 480 5.30.05919 1.416 315 1.30.00585 1.6362>10.00.00057 1.543 255 2.10.01222 1.495 116 6.20.01779 2.376 953 2.30.03563 1.2844>10.00.00016 2.2160 5.10.006940.708137.90.00319 2.303 3283 3.40.34957 1.353 2798.70.02859 1.299 773 3.90.057280.943 493 5.30.04212 1.304 11447.10.090460.92115617.50.124780.98 259 5.70.03346 1.295 372 2.80.02555 1.219 1289>10.00.12482 1.03 7277.10.06585 1.03830 5.80.00343 1.39794 6.80.01439 1.685 303 6.60.019140.892 2119 6.70.21487 1.164 9987.10.059110.76263 3.40.002370.436140.000190.962 475 5.50.03101 1.1898.60.00225 1.225 194 2.50.00680.908 817 6.60.031350.985 68170.09849 1.714 1358 2.50.073 1.334 1194 2.60.032550.749 10720.00309 1.26 1021 4.40.12987 1.194 459 3.20.02416 1.192 2618.90.03446 1.396 1804 3.70.15067 1.296 1418.60.01298 1.609 1709 4.50.08330.842 327 1.30.00320.94181 4.90.00904 1.23 327 4.70.03579 1.258 17750.0131 1.044 2597.10.02119 1.078 877 4.30.068460.99799 1.40.001410.915 449 6.30.028910.786 16157.50.18356 1.081 22860.020170.96535 1.90.00176 1.224 1160 3.80.055440.914 201 6.30.017610.949 426 4.10.025740.772 2120 3.70.074070.708 1016 5.80.054290.757 321 5.50.013530.63690 3.30.006210.726 184 4.80.010070.73123>10.00.00228 1.513 610 3.80.032170.72 842 5.40.057730.829 213 5.40.01270.77439 2.40.001731002 4.80.033670.729 208 4.30.00930.537 5247.60.043570.93 116>10.00.02104 1.812 173>10.00.013330.936 589 5.30.032210.807 108 6.10.009190.931 487 4.70.03951 1.142 76450.047770.779 1666 5.20.091160.854 193 1.80.004620.921 120 2.20.004670.894 3338.70.027140.756 4327.10.027040.625 19520.003610.533 194 6.90.011710.909 2597.30.013360.716 266 6.80.026850.752 87080.070140.869 2187.70.01310.75610 5.60.00062 1.012 4189.20.026590.815 622 5.20.034970.666 1709.90.01712 1.222 178 5.80.009120.836 907.70.00320.65258>10.00.01221 1.371 4877.30.041850.896 187 5.60.009820.662 338 6.50.019280.711 443 6.30.026840.651 20850.01330.806 409 4.50.027060.735 774 3.20.034350.762 15287.90.107180.745 739 5.40.053960.715 227.60.001180.774 105 5.10.007480.647 129 4.90.00780.74475 3.50.002990.639212 3.70.013920.784 201 5.60.009290.599 500 6.30.06138 1.238 >10.00.00042 1.049 290 6.10.019640.625 251 5.60.017330.895 12498.40.075460.68 241 2.20.005560.628 1341 6.70.120620.829 651 6.90.026690.572 224 5.10.010210.672 290 5.30.013220.702 194 1.60.002660.7278>10.00.000620.792 314>10.00.019270.649 32830.001390.381 520 6.30.034550.62153 5.20.004350.904 200 1.30.002460.856 109 3.70.00563143 5.80.013670.825 439 4.20.025470.66 441>10.00.03210.853 17290.80.002020.494 1527.40.009010.741 624 2.40.016810.714 337 6.40.02850.71 173 1.90.002010.614 124 6.30.008010.593 2207.60.010810.602 4708.20.028730.596 221 5.20.00853 1.29 4957.50.036490.714 967.20.003150.52311>10.00.000450.543 184 3.50.006730.588 1064 2.80.017560.666 2437.60.014140.744 257.10.002380.868 105130.022770.574 10130.003550.834 2518.20.017620.63 16669.90.081330.508 1478 4.10.10840.548 14830.008560.8753427.80.015630.587 559.80.002470.6496 1.50.001250.668648.10.002270.561 178 6.20.010850.6 247>10.00.011750.526 202 5.20.013550.701 1484 5.40.069550.546 116 6.40.005740.60572 2.10.00140.58954>10.00.004010.856 351 6.60.011110.57 960 3.90.021660.681 1549.50.006150.597 5790.008180.646 6720.0008353 2.30.00061176 2.60.005320.633 1947.80.016490.569 223 5.60.014960.968 25280.006360.457 1626 5.50.049760.537 1977.10.008730.585 128 5.40.00580.57919>10.00.001080.734 650 3.70.017210.673 9419.10.050830.59113>10.00.00130.598 21090.007690.475 906>10.00.066080.687 350 3.90.005590.396 2247.80.01640.479 1789 5.50.074220.539 419>10.00.014550.491 579 6.80.034280.585 11717.30.084950.73 918.60.003920.471 458.60.00220.655 100 6.20.007370.666 335 5.10.012850.644 964 5.50.038290.59199 5.20.004810.478 302 6.60.012460.48640 2.10.000610.345 392>10.00.021350.719339>10.00.01560.489 5138.90.025150.615 50 4.50.001620.659 1368.50.005580.639 353 5.90.018730.713 495 4.80.011170.321 1147.60.003960.391 156 6.70.022520.706 508 6.50.027410.476 82 5.10.003380.472 410>10.00.020760.421 400>10.00.012770.531 463 4.70.008250.53 728 5.70.012880.253 2367.10.012090.669 180 3.80.004490.45 360>10.00.018270.679 2957.80.035210.653 950 3.40.026580.586 218 5.20.006840.446 2547.50.007820.487。
7. Endotoxin LALTests
Charles River Endosafe
2
Woo Jung BSC Inc.
August 25, 2003
LAL Discoveries by Bang and Levin
Described role of endotoxin in coagulation of Limulus blood
Prepared Endotoxin - responsive lysate from Amoebocytes
Endotoxicity
ENDOTOXIN CAUSES HUMAN TISSUE TO RELEASE INFLAMMATORY MEDIATORS INFLAMMATION INDUCES A VARIETY OF TISSUE DAMAGE SHOCK and MULTIPLE ORGAN DYSFUNCTION MAY OCCUR
Endotoxins and Pyrogens
Pyrogens are fever-inducing agents in humans and animals
include endotoxin, gram + cell debris, fungi
Endotoxins are components from the outer membrane of gram-negative bacteria
Clotting Enzyme
Liquid Coagulogen
M++ pH=7.2
Clotted Coagulin Gel
Summary of Gel Clot Test
Endpoint sought by 180 inversion of sample tube
达格列净联合二甲双胍治疗初诊2_型糖尿病的临床疗效分析
达格列净联合二甲双胍治疗初诊2型糖尿病的临床疗效分析卢宁，刘艳梅，仇颖盐城市第一人民医院内分泌科，江苏盐城224000[摘要]目的探讨达格列净联合二甲双胍治疗初诊2型糖尿病患者的临床疗效及其安全性。
方法选取2021年9月—2022年11月在盐城市第一人民医院内分泌科接受治疗的初诊2型糖尿病患者70例为研究对象，根据系统盲选分组法分成两组，每组35例。
对照组应用二甲双胍治疗，观察组应用达格列净+二甲双胍治疗。
检测并对比两组患者血糖指标、血脂指标、体质指数、不良反应发生情况及疗效。
结果治疗后，观察组血糖及血脂各项指标水平均优于对照组，体质指数低于对照组，差异有统计学意义（P<0.05）。
观察组治疗总有效率高于对照组，差异有统计学意义（P<0.05）。
两组不良反应发生率比较，差异无统计学意义（P>0.05）。
结论达格列净联合二甲双胍对初诊2型糖尿病患者具有良好的治疗作用，有助于改善血糖和血脂、体质量水平，不良反应较少。
[关键词] 达格列净；二甲双胍；初诊2型糖尿病；不良反应；治疗效果[中图分类号] R587.1 [文献标识码] A [文章编号] 1672-4062（2023）03（b）-0091-04 Clinical Analysis of Dapagliflozin Combined with Metformin in the Treat⁃ment of Newly Diagnosed Type 2 Diabetes MellitusLU Ning, LIU Yanmei, QIU YingDepartment of Endocrinology, Yancheng First People´s Hospital, Yancheng, Jiangsu Province, 224000 China[Abstract] Objective To investigate the clinical efficacy and safety of Dapagliflozin combined with metformin in the treatment of newly diagnosed type 2 diabetes patients.Methods Seventy newly diagnosed patients with type 2 diabetes who received treatment in the Endocrinology Department of Yancheng First People´s Hospital from September 2021 to November 2022 were selected as the research objects and divided into two groups with 35 cases in each group accord⁃ing to systematic blind selection grouping. The control group was treated with metformin, and the observation group was treated with dagliazine+metformin. Blood glucose index, blood lipid index, body mass index, occurrence of ad⁃verse reactions and efficacy of the two groups were detected and compared.Results After treatment, the blood glucose and lipid levels of the observation group were better than those of the control group, while the body mass index of the observation group was lower than that of the control group, the difference was statistically significant (P<0.05). The to⁃tal effective rate of the observation group was higher than that of the control group, and the difference was statistically significant (P<0.05). There was no statistically significant difference in the incidence of adverse reactions between the two groups (P>0.05).Conclusion Dapagliflozin combined with metformin has a good therapeutic effect on newly diag⁃nosed type 2 diabetes patients, which can help to improve blood glucose, blood fat and body mass level, and less ad⁃verse reactions.[Key words] Dapagliflozin; Metformin; Newly diagnosed type 2 diabetes mellitus; Adverse reaction; Therapeutic effect目前，老年化问题越发突出，加之国民饮食习惯、膳食结构以及饮食偏好的变化，使得糖尿病患者逐渐增多[1]。
GC测定盐酸普拉克索中三乙胺残留量
WHO International Standard 1st WHO International Standard for Human Papillomavirus (HPV) Type 16 DNA
WHO International Standard1st WHO International Standard for Human Papillomavirus (HPV)Type 16 DNA NIBSC code: 06/202 Instructions for use(Version 2.0, Dated 10/11/2010)1. INTENDED USEThe 1st International Standard for HPV Type 16 (HPV-16) DNA Nucleic Acid Amplification Techniques consists of a freeze-dried preparation of recombinant plasmid containing full-length HPV-16 DNA cloned via its unique BamH1 site (Quint et al., 2006). The standard has been formulated in a background of purified human genomic DNA, lyophilized in 0.5 ml aliquots and stored at -20 °C. The material was calibrated in an international collaborative study involving 19 laboratories (Wilkinson et al., 2008). The International Standard contains material that is proprietory to third parties and should be used for the sole purpose of calibrating in-house or working standards for the amplification and detection of HPV-16 DNA. The International Standard should not be used for any other purpose and should be discarded after use. 2. CAUTIONThis preparation is not for administration to humans .This material contains DNA derived from C33A cells. As with all materials of biological origin, this preparation should be regarded as potentially hazardous to health. It should be used and discarded according to your own laboratory's safety procedures. Such safety procedures should include the wearing of protective gloves and avoiding the generation of aerosols. Care should be exercised in opening ampoules or vials, to avoid cuts.3. UNITAGEThe 1st International Standard for HPV-16 DNA Nucleic Acid Amplification Techniques has been assigned a unitage of 5 x 106 International Units (IU) per ampoule.Traceability statement:It was proposed at a WHO meeting in January 2008 (WHO Meeting Report, 2008) that the instructions for use of the International Standard for HPV-16 DNA include the calculations and assumptions used in determining the theoretical HPV-16 qenome equivalents (GEq) of the bulk material used in formulating the International Standard, thus demonstrating that 1 IU is equivalent to 1 GEq for HPV-16 DNA . The definitive unitage of the 1st WHO International Standard for HPV-16 DNA therefore remains as IU while the traceability statement would allow users to equate IU with GEq.Assays for DNA concentration of the recombinant HPV-16 plasmid stock preparation were performed in Dr Cosette Wheeler‟s laboratory, University of New Mexico (UNM). DNA concentrations were determined by absorbance at 260 nm as well as spectrofluorometrically using the Picogreen assay (Invitrogen Corporation, USA). A correlation coefficient of 0.95 or higher was obtained between the two DNA measurements. 10 ng HPV-16 plasmid DNA/μl was supplied to NIBSC for formulating the bulk material for subsequent freeze-drying. The UNM laboratory also provided NIBSC with a statement indicating that 1.0 x 1011 GEq/ml for HPV-16 is equal to 1.17 ng/μl. 10 ng HPV-16 plasmid DNA/μl plasmid stock preparation is therefore equivalent to 8.547 x 1011 HPV-16 GEq/ml. NIBSC used this data in formulating the 1st International Standard for HPV Type 16 DNA.Formulation of bulk material for the 1st International Standard for HPV Type 16 DNA (NIBSC code 06/202):At NIBSC, the bulk HPV-16 plasmid DNA material was prepared according to the formula:HPV GEq/ml of bulk material = (HPV GEq/ml of plasmid stock x volume plasmid stock) / volume bulk material.Therefore,HPV-16 GEq/ml of bulk material = (8.547 x 1011 HPV-16 GEq/ml plasmid stock) x (0.02223 ml HPV-16 plasmid stock) / 1900 ml HPV-16 bulk material = 1.0 x 107 HPV-16 GEq/ml bulk materialThe HPV-16 DNA bulk material was subsequently freeze-dried in 0.5 ml aliquots.Certain assumptions are required for equating IU to GEq for the 1st International Standard for HPV-16 DNA: 1) 1.0 x 1011 GEq/ml for HPV-16 is equal to 1.17 ng/μl. 2) There is no loss in activity of the HPV-16 DNA upon lyophilization. 3) The recombinant HPV-16 plasmid DNA accurately mimics the activity of HPV-16 viral DNA in biological samples.Independent calculation of GEq/ml for recombinant HPV-16 plasmid DNA.NIBSC also independently calculated the genome equivalence of the HPV-16 plasmid stock preparation and bulk preparation in which the molecular weights of the full-length HPV-16 genome and pBR322 DNA were based on sequence content using BioEdit Sequence Alignment Editor v7.0.5.3 (Tom Hall, Isis Pharmaceuticals Inc., USA). The sequences used for determining the molecular weights are GenBank Accession number J01749.1 for pBR322 and the reference sequence for HPV16 (Accession K02718).BioEdit dataDNA molecule: HPV16 Accession K02718 Length = 7904 base pairsMW= 4786756.00 Daltons, double strandedDNA molecule: cloning vector pBR322 Length = 4361 base pairsMW= 2653867.00 Daltons, double strandedFormulaeGEq/ml of the HPV plasmid stock was calculated according to the formula: GEq/ml of the HPV plasmid stock = (DNA concentration of HPV plasmid stock) x (MW of HPV DNA + MW of pBR322)-1 x (Avogadro‟s Number) where Avogadro‟s Number = 6.022x1023 molecules/molGEq/ml of the bulk HPV DNA materials was calculated according to the formula:HPV GEq/ml of bulk material = (HPV GEq/ml of plasmid stock x volume plasmid stock) / volume bulk material.CalculationThe recombinant HPV-16 plasmid stock preparation was supplied to NIBSC at a concentration of 10 ng/μl. Using the MW determinations shown above, the GEq/ml of the HPV-16 plasmid stock is:= (10 x 10-9 g/μl) x (mol/(7440623 g) x (6.022x1023 molecules/mol) = 8.093 x 108 molecules/μl = 8.093 x 1011molecules/ml = 8.093 x 1011 HPV-16 GEq/ml22.23μl of the recombinant HPV-16 plasmid stock was diluted to a final volume of 1900ml, therefore,HPV-16 GEq/ml of bulk material = (8.093 x 1011 HPV-16 GEq/ml plasmid stock) x (0.02223 ml HPV-16 plasmid stock) / 1900 ml HPV-16 bulk material = 0.947 x 107 HPV-16 GEq/ml bulk material4. CONTENTSCountry of origin of biological material: United Kingdom.Each ampoule contains the lyophilized equivalent of 0.5 ml HPV-16 plasmid DNA in 10mM Tris buffer pH7.4 containing 1mM EDTA, 5 mg/ml trehalose and ~1 x 106 human GEq/ml derived from C33a cells.5. STORAGEThe ampoule should be stored at -20 °C or below on receipt.Please note: because of the inherent stability of lyophilized material, NIBSC may ship these materials at ambient temperature.6. DIRECTIONS FOR OPENINGDIN ampoules have an …easy -open‟ coloured stress point, where the narrow ampoule stem joins the wider ampoule body.Tap the ampoule gently to collect the material at the bottom (labeled) end. Ensure that the disposable ampoule safety breaker provided is pushed down on the stem of the ampoule and against the shoulder of the ampoule body. Hold the body of the ampoule in one hand and the disposable ampoule breaker covering the ampoule stem between the thumb and first finger of the other hand. Apply a bending force to open the ampoule at the coloured stress point, primarily using the hand holding the plastic collar.Care should be taken to avoid cuts and projectile glass fragments that might enter the eyes, for example, by the use of suitable gloves and an eye shield. Take care that no material is lost from the ampoule and no glass falls into the ampoule. Within the ampoule is dry nitrogen gas at slightly less than atmospheric pressure. A new disposable ampoule breaker is provided with each DIN ampoule.7. USE OF MATERIALNo attempt should be made to weigh out any portion of the freeze-dried material prior to reconstitution.The 1st International Standard for HPV-16 DNA contains high copy number template. There is a high risk of HPV-16 plasmid DNA contamination via aerosolization upon opening of the glass ampoule. The material must be opened and handled in a separate laboratory environment, away from other pre-amplification components such as reagents, labware and samples.The material is supplied lyophilized and, before use, should be reconstituted in 0.5 ml sterile nuclease-free water. Ensure that the inside surface of the ampoule is wetted with the added water so that any particles of freeze-dried material adhering to the glass are reconstituted. The reconstituted material has a final concentration of 1 X 107 IU/ml. The reconstituted material is suitable for calibration of in-house or working standards for the amplification and detection of HPV-16 DNA.. The material is not suitable for calibrating or assessing extraction, precipitation or centrifugation procedures. The material has NOT been calibrated for human DNA nucleic acid amplification techniques.8. STABILITYReference materials are held at NIBSC within assured, temperature-controlled storage facilities. The 1st International Standard for HPV-16 DNA should be stored at -20 °C or below on receipt.Studies on the stability of reconstituted standard are underway. Users should determine the stability of the reconstituted material according to their own method of preparation, storage and use.NIBSC follows the policy of WHO with respect to its reference materials.9. REFERENCESQuint, W. G. V., Pagliusi, S. R., Lelie, N., de Villiers, E. M., Wheeler, C. M. and the World Health Organization Human Papillomavirus DNA International Collaborative Study Group. (2006). Results of the First WorldHealth Organization International Collaborative Study of Detection of Human Papillomavirus DNA. J. Clin. Microbiol. 44: 571-579.Wilkinson, D.E., Baylis, S.A., Padley, D., Heath, A.B., Ferguson, M., Pagliusi, S.R., et al. Establishment of the 1st World Health Organization international standards for human papillomavirus type 16 DNA and type 18 DNA. Int J Cancer 2010 Jun 15;126(12):2969-83.WHO meeting report, on “Standardization of HPV assays and the role of HPV LabNet in supporting vaccine introduction” Geneva, Switzerland, 23-25 January 2008, in preparation.10. ACKNOWLEDGEMENTS11. FURTHER INFORMATIONFurther information can be obtained as follows; This material: enquiries@ WHO Biological Standards:http://www.who.int/biologicals/en/JCTLM Higher order reference materials: /en/committees/jc/jctlm/ Derivation of International Units:/products/biological_reference_materials/frequently _asked_questions/how_are_international_units.aspx Ordering standards from NIBSC:/products/ordering_information/frequently_asked_q uestions.aspxNIBSC Terms & Conditions:/terms_and_conditions.aspx12. CUSTOMER FEEDBACKCustomers are encouraged to provide feedback on the suitability or use of the material provided or other aspects of our service. Please send any comments to enquiries@13. CITATIONIn all publications, including data sheets, in which this material is referenced, it is important that the preparation's title, its status, the NIBSC code number, and the name and address of NIBSC are cited and cited correctly.15. LIABILITY AND LOSSInformation provided by the Institute is given after the exercise of all reasonable care and skill in its compilation, preparation and issue, but it is provided without liability to the Recipient in its application and use. It is the responsibility of the Recipient to determine the appropriateness of the standards or reference materials supplied by the Institute to the Recip ient (“the Goods”) for the proposed application and ensure that it has the necessary technical skills to determine that they are appropriate. Results obtained from the Goods are likely to be dependant on conditions of use by the Recipient and the variability of materials beyond the control of the Institute.All warranties are excluded to the fullest extent permitted by law, including without limitation that the Goods are free from infectious agents or that the supply of Goods will not infringe any rights of any third party.The Institute shall not be liable to the Recipient for any economic loss whether direct or indirect, which arise in connection with this agreement.The total liability of the Institute in connection with this agreement, whether for negligence or breach of contract or otherwise, shall in no event exceed 120% of any price paid or payable by the Recipient for the supply of the Goods.If any of the Goods supplied by the Institute should prove not to meet their specification when stored and used correctly (and provided that the Recipient has returned the Goods to the Institute together with written notification of such alleged defect within seven days of the time when the Recipient discovers or ought to have discovered the defect), the Institute shall either replace the Goods or, at its sole option, refund the handling charge provided that performance of either one of the above options shall constitute an entire discharge of the Institute‟s liability under this Condition.。
血管外膜细胞钙化及其钙化机制研究
血管外膜细胞钙化及其钙化机制研究谭小青,张旭升,樊小容,黄战军摘要 目的:研究经体外诱导钙化建立大鼠血管外膜细胞钙化模型,检测钙化过程中成骨相关指标及凋亡㊁自噬相关蛋白的表达变化,旨在为心血管疾病模型提供更精确的细胞模型,并初步探讨其钙化机制㊂方法:原代提取大鼠胸主动脉外膜纤维细胞,取3~6代细胞使用诱导培养基(高糖DMEM +10%胎牛血清+10mmol/L β-甘油磷酸+0.05mmol/L 抗坏血酸+100mmol/L 地塞米松)诱导钙化,诱导时间为3d ㊁6d ㊁9d ㊁12d ㊁15d ,筛选出诱导细胞钙化的最佳时间㊂对细胞采用茜素红S 染色㊁细胞内钙含量测定和碱性磷酸酶(ALP )活性检测,鉴定是否成功构建钙化模型㊂采用实时定量聚合酶链式反应(PT -PCR )检测成骨相关因子骨形态生成蛋白2(BMP2)和核心结合因子α1(Runx2)的mRNA 含量,蛋白免疫印迹法(Western Blot )检测凋亡蛋白Bax ㊁Bcl -2和自噬相关蛋白微血管相关蛋白(LC3)㊁Beclin -1的表达水平,找出血管外膜细胞钙化的潜在机制㊂结果:当诱导钙化时间为15d 时,血管外膜细胞中主要钙化指标胞内钙含量及ALP 活性上调(P <0.05),茜素红S 染色显示钙化组有明显钙盐沉积㊂血管外膜细胞经钙化诱导后,BMP2和Runx2的mRNA 水平上调,Bax 蛋白水平上调,Bcl -2和Beclin -1蛋白水平下调,LC3-Ⅱ/LC3-Ⅰ比值上调(P <0.05)㊂结论:钙化诱导培养基培养血管外膜细胞15d 可成功构建钙化细胞模型,血管外膜细胞钙化可能与细胞向成骨样表型转化有关,血管外膜细胞钙化过程涉及细胞自噬及凋亡调控㊂关键词 血管外膜细胞;钙化;成骨样表型转化;自噬与凋亡;实验研究d o i :10.12102/j.i s s n .1672-1349.2023.18.010 Calcification of Vascular Adventitial Cells and Its MechanismTAN Xiaoqing,ZHANG Xusheng,FAN Xiaorong,HUANG Zhanjun Longgang District People 's Hospital of Shenzhen,Shenzhen 518172,Guangdong,China Corresponding Author ZHANG Xusheng,E -mail:*****************Abstract Objective:To investigate the mananism of calcification of rat vascular adventitial cells,establish the calcification model of rat vascular adventitial cells,and detect the expression changes of osteogenesis -related indicators,apoptosis,and autophagy -related proteins during the calcification process.It aimed to provide more accurate cell models for cardiovascular disease and initially explore the mechanism of calcification.Methods:Rat thoracic aortic adventitial fibroblasts were extracted from the primary generation,and the 3rd to 6th generation cells were used for induction medium(high glucose DMEM +10%fetal bovine serum +10mmol/L β-glycerophosphate +0.05mmol/L ascorbic acid +100mmol/L dexamethasone)to induce calcification,the induction time was 3,6,9,12,and 15d,and the optimal time for inducing cell calcification was selected.The cells were stained with alizarin red S,detected by intracellular calcium content and alkaline phosphatase(ALP)to identify whether the calcification model was successfully constructed.Real -time quantitative reverse transcription polymerase chain reaction(RT -PCR)was used to detect the mRNA levels of osteogenesis -related factors bone morphogenetic protein(BMP2)and runt -related transcription factor 2(Runx2);Western Blot was used to detect the apoptosis proteins Bax,Bcl -2,the autophagy -related proteins LC -3,and Beclin -1expression level;then the potential mechanism of vascular adventitial cell calcification would be revealed.Results:When calcification was induced for 15days,the intracellular calcium content in the adventitial cells of the main calcification indicators and ALP activity were up -regulated(P <0.05).Alizarin red S staining showed obvious calcium deposits in the calcification group.After calcification was induced in adventitial cells,the mRNA levels of BMP2and Runx2up -regulated,the protein levels of Bax up -regulated,the protein levels of Bcl -2and Beclin -1down -regulated,and the ratio of LC3-Ⅱ/LC3-Ⅰdown -regulated(P <0.05).Conclusion:Adventitial cells cultured in the calcification -inducing medium for 15days could successfully construct a calcified cell model.calcification of adventitial cells might be related to the transformation of cells to an osteoblast -like phenotype.The Calcification process of adventitial cells involved autophagy and apoptosis regulation.Keywords adventitial cells;calcification;osteogenic phenotype transformation;autophagy and apoptosis;experimental study血管钙化常见于动脉粥样硬化㊁血脂异常㊁高血压㊁糖尿病㊁慢性肾病及衰老等人群[1],血管钙化引起血管硬度增加㊁顺应性降低,导致心肌缺血㊁心力衰竭㊁血栓形成等,增加脑卒中㊁心脏病㊁动脉粥样硬化斑块破裂等的风险,被认为是影响心血管疾病的重要因素之一[2-4]㊂目前关于血管内膜㊁中膜和心脏瓣膜钙化的关注和研究相对较多㊂临床工作中发现,血管外膜也可发生钙化,然而调查发现,现阶段对血管外膜钙化的作者单位 深圳市龙岗区人民医院(广东深圳518172)通讯作者 张旭升,E -mail :*****************引用信息 谭小青,张旭升,樊小容,等.血管外膜细胞钙化及其钙化机制研究[J ].中西医结合心脑血管病杂志,2023,21(18):3347-3350.关注较少,因此,需要更多的研究来阐明血管钙化的致病机制㊂最初血管钙化被认为是被动和退行性病变,标志着血管老化,但是越来越多研究表明血管钙化是类似于胚胎骨形成的病理生物学过程[5-6]㊂Bostr öm 等[7-8]研究发现,钙化过程中大鼠血管中膜细胞由原有收缩表型转变成为成骨样细胞表型,原有的收缩标志物如平滑肌肌动蛋白α(α-SMA )等表达减少,并表达核心结合因子α1(Runx2)㊁骨形态生成蛋白2(BMP2)等多种成骨样标志物,从而介导骨基质在血管中沉积㊂细胞凋亡与自噬为2种细胞死亡的方式,与血管钙化息息相关,研究表明,血管中膜细胞在细胞凋亡过程中释放凋亡小体,促进细胞钙化,而细胞自噬通过多种机制调控细胞钙化[9-10]㊂本研究对大鼠血管外膜细胞进行体外诱导钙化,建立大鼠血管外膜细胞钙化模型,并检测钙化过程中成骨相关指标及凋亡㊁自噬相关蛋白的表达变化,旨在为心血管疾病模型提供更精确的细胞模型,并初步探讨其钙化机制㊂1材料与方法1.1试剂胎牛血清(FBS,Gibco),青霉素,链霉素(Gibco,美国),茜素红S溶液,β-甘油磷酸,抗坏血酸,地塞米松(Sigma,美国),抗GAPDH抗体(Bioworld),抗Bcl-2, Bax,Bcelin1和微血管相关蛋白(LC3)抗体(CST),碱性磷酸酶检测试剂盒㊁钙(Ca)检测试剂盒(南京建城生物工程研究所)㊂1.2大鼠血管外膜细胞分离与培养取10只4~6周龄雄性Wistar-Kyoto大鼠(体质量120~180g)胸主动脉分离血管外膜,采用组织黏附法培养㊂使用添加10%胎牛血清的高糖DMEM培养基(Gibco dmem)在37ħ㊁5%二氧化碳条件下培养细胞㊂当细胞增殖至80%~90%融合时,用0.25%胰酶消化传代㊂使用第3代至第6代的细胞进行后续实验㊂1.3体外钙化模型的建立钙化诱导培养基为含10%胎牛血清,10mmol/L β-甘油磷酸钠,0.05mmol/L抗坏血酸和100mmol/L 地塞米松的高糖DMEM培养液㊂将第3代至第6代细胞分为对照组和钙化组,待细胞长至50%融合时,使用钙化诱导培养基培养,每3d更换1次培养基,连续培养15d㊂1.4碱性磷酸酶(ALP)酶活测定细胞钙化诱导后,弃去培养基,1ˑ磷酸缓冲盐溶液(PBS)洗细胞3次,加入裂解液500μL(1%T ritonX-100),冰上裂解40min后,离心,取上清液㊂使用上清液根据试剂盒说明书检测ALP活性及总蛋白含量㊂1.5细胞内钙含量检测细胞钙化诱导后,弃去培养基,1ˑPBS洗细胞3次,每孔加入500μL0.6mol/L的盐酸4ħ脱钙过夜,取上清,根据钙测试试剂盒说明书检测钙含量㊂将脱钙后的细胞用4ħPBS洗3次,每孔加入500μL NaOH/0.1%SDS裂解细胞,取上清,用二喹啉甲酸法(BCA)测定细胞蛋白含量㊂1.6茜素红S染色细胞钙化诱导15d,弃去培养基,1ˑPBS洗细胞3次,加入0.5mL4%多聚甲醛室温固定15min,用双蒸水洗3次,加入1mL0.1%茜素红室温孵育15min,吸去染液,双蒸水洗3次,在倒置显微镜下观察㊂1.7实时定量聚合酶链式反应(RT-PCR)检测细胞钙化诱导后,弃去培养基,1ˑPBS洗细胞3次,使用TaKaRa MiniBEST Universal RNA Extraction Kit提取总RNA,使用PrimeScrip TM RT reagent Kit将所提取的RNA逆转录合成cDNA,以cDNA为模板,通过SYBR Green I嵌合荧光定量RT-PCR检测BMP-2㊁Runx2和GAPDH的表达量㊂引物序列见表1㊂表1引物序列基因方向序列Runx2正向5'-TGGCTTTGGTTTCAGGTTAGG-3'反向5'-TGGAGATGTTGCTCTGTTCG-3' BMP-2正向5'-TGAGGATTAGCAGGTCTTTGC-3'反向5'-TCTCGTTTGTGGAGTGGATG-3' GAPDH正向5'-GGCTGCCCAGAACATCAT-3'反向5'-CGGACACATTGGGGGTAG-3'1.8蛋白免疫印迹法(Western Blot)检测细胞钙化诱导15d,弃去培养基,1ˑPBS洗细胞3次,提取细胞总蛋白㊂使用12%SDS-PAGE胶电泳分离,并转移到聚偏二氟乙烯膜(PVDF)上,封闭后,加入一抗(Bax1ʒ1000,Bcl-21ʒ1000,Beclin11ʒ1000, LC31ʒ1000,GAPDH1:1000)稀释液,4ħ孵育过夜;加入二抗稀释液(1ʒ10000)室温孵育1h后,使用ECL发光试剂盒显影并计算灰度值㊂1.9统计学处理应用SPSS19.0软件进行统计处理,符合正态分布的定量资料以均数ʃ标准差(xʃs)表示,比较采用t检验,以P<0.05为差异有统计学意义㊂2结果2.1大鼠血管外膜细胞可在体外被诱导钙化为验证高磷是否能诱导大鼠血管外膜细胞钙化,使用钙化诱导培养基培养细胞,在不同时间点检测ALP活性和胞内钙含量㊂随着培养时间延长,ALP活性逐渐上升,在培养第12天达到峰值,与对照组比较差异有统计学意义(P<0.05);诱导第3天开始所测得的胞内钙含量与对照组比较升高(P<0.05),ALP 活性和钙含量升高具有时间依赖性㊂详见图1㊁图2㊂诱导15d所测得钙含量最高,因此,后续实验选择的诱导时间为15d㊂对钙化诱导15d的细胞进行茜素红S染色,结果显示,对照组细胞呈长梭形,而钙化组细胞变成菱形㊂茜素红S染色后,钙化组可观察到大量的橘红色钙结节(见图3),而对照组完全没有㊂这也证明大鼠血管外膜细胞可在体外被钙化培养基诱导钙化㊂图1钙化诱导培养基诱导外膜细胞后ALP含量(与0d时比较,*P<0.05)图2钙化诱导培养基诱导外膜细胞后胞内钙含量(与0d时比较,*P<0.05)图3培养15d时细胞经茜素S红染色切片图(ˑ100)2.2血管外膜细胞钙化与细胞向成骨样表型转化有关血管钙化的增加与成骨细胞特异性标志物如BMP2㊁和Runx2的增加有关[11]㊂RT-PCR结果显示,与对照组比较,钙化组的成骨细胞特异性标志物BMP2和Runx2mRNA表达量增加,与对照组比较差异有统计学意义(P<0.05)㊂详见图4㊂图4外膜细胞钙化过程中BMP2和Runx2mRNA表达量(与对照组比较,*P<0.05)2.3血管外膜细胞钙化过程涉及细胞自噬及凋亡调控通过Western Blot检测凋亡和自噬相关蛋白的表达量变化㊂与对照组比较,钙化组促凋亡蛋白Bax表达上调,抑凋亡蛋白Bcl-2表达下调(P<0.05)㊂详见图5㊂钙化组自噬相关蛋白Beclin1表达上调,LC3-Ⅱ/ LC3-Ⅰ比例上调(P<0.05),说明钙化诱导培养后细胞内凋亡水平上调㊁自噬水平升高㊂详见图6㊂图5诱导钙化后促凋亡蛋白及抑凋亡蛋白表达变化图6诱导钙化后凋亡及自噬蛋白Beclin1等表达变化3讨论血管钙化作为心血管疾病病人的并发症之一,其发病率与严重程度逐年增高及加重,是导致心血管疾病病人高死亡率的重要因素㊂血管钙化缺乏有效的治疗药物㊂因此,探究血管钙化发病机制,在分子水平寻找有效的诊断和防治靶点是急需开展的基础研究工作㊂本研究证明,使用10mmol/Lβ-甘油磷酸+0.05 mmol/L抗坏血酸+100mmol/L地塞米松培养外膜细胞即可诱导大鼠血管外膜细胞在体外发生钙化,这是通过茜素红S染色㊁ALP活性检测及胞内钙含量检测结果得以确定的㊂血管钙化过程中,血管中膜细胞向成骨样细胞表型转变并表达相关成骨标志物,从而引起骨基质的沉积,是血管钙化的重要特点及机制[5]㊂本实验所用的血管外膜细胞钙化条件与血管中膜细胞钙化条件一致,说明血管外膜细胞钙化的机制可能与中膜细胞钙化的机制部分一致㊂血管中膜细胞钙化过程中,细胞表达成骨相关的转录因子如Runx2等,进而促进下游表达骨相关蛋白如骨形态发生蛋白BMP2等的表达,从而促使细胞向成骨样细胞主动分化[12-13],本研究也观察到类似的机制㊂通过PT-PCR检测,发现钙化培养基培养大鼠血管外膜细胞15d后,BMP2和Runx2的mRNA表达水平升高㊂本研究通过对钙盐沉积与成骨样细胞表型转变2个维度的探讨,证明血管外膜细胞可在体外被诱导钙化,丰富了血管钙化的分型㊂血管钙化的发生机制复杂,涉及多种信号通路,如细胞自噬和凋亡㊁Wnt/β-catenin信号通路激活㊁内质网应激等均参与调控血管钙化的过程㊂自噬作为一种细胞应激的适应性反应,在维持血管结构与功能中十分关键㊂研究表明,血管钙化过程中自噬水平增高[14-15]㊂在体外实验中,高磷可提高大鼠血管中膜细胞的自噬水平,增加细胞内自噬体数量,从而抑制凋亡与钙化[16]㊂还有研究表明,自噬可通过抑制大鼠血管中膜细胞氧化应激,抑制血管内皮细胞的炎症反应,对三酰甘油等脂代谢进行调控,从而减轻血管钙化[17-18]㊂LC3和Beclin1是2种典型的自噬标志物,Western Blot实验结果表明,用钙化培养基诱导大鼠血管外膜细胞15d,LC3-Ⅱ/LC3-Ⅰ比率升高,Beclin1蛋白水平表达升高,说明细胞内自噬水平升高㊂多项研究表明,细胞凋亡参与促进血管钙化的发生,抑制细胞凋亡和抑制钙化[16-17]㊂在对大鼠的体内研究发现,成纤维细胞生长因子21通过内质网应激调控Caspase-12信号通路来减少血管内中膜细胞凋亡,从而抑制血管钙化[18]㊂另外,提高培养基中的Pi 或Ca2+浓度,可诱导细胞质膜形成并释放基质囊泡(如凋亡小体),从而导致细胞外基质钙化,这种基质钙化可能成为血管钙化的成核位点[19]㊂Bax和Bcl-2是2种典型的凋亡和抑制凋亡蛋白,本实验结果证明,利用钙化培养基对血管外膜细胞诱导钙化过程中,细胞内凋亡水平升高㊂同时细胞内自噬水平也升高,这可能是细胞自我调控以对抗钙化的结果㊂本研究证实血管外膜细胞可在体外被诱导钙化,且外膜钙化过程与骨组织钙化过程类似,为主动可调控的过程㊂血管钙化是一个复杂的过程,涉及细胞凋亡和自噬等调控通路,仍需进一步研究㊂参考文献:[1]梁英权,段亚君,韩际宏.血管钙化分子机制研究进展[J].中国动脉硬化杂志,2020,28(11):921-929.[2]NICOLL R,HENEIN M Y.The predictive value of arterial andvalvular calcification for mortality and cardiovascular events[J].Int J Cardiol Heart Vessel,2014,3:1-5.[3]JOHNSON R C,LEOPOLD J A,LOSCALZO J.Vascularcalcification:pathobiological mechanisms and clinical implications[J].Circulation Research,2006,99(10):1044-1059.[4]YAMADA S,GIACHELLI C M.Vascular calcification in CKD-MBD:roles for phosphate,FGF23,and Klotho[J].Bone,2017,100:87-93.[5]LIN M E,CHEN T M,WALLINGFORD M C,et al.Runx2deletion insmooth muscle cells inhibits vascular osteochondrogenesis andcalcification but not atherosclerotic lesion formation[J].Cardiovascular Research,2016,112(2):606-616.[6]DURHAM A L,SPEER M Y,SCATENA M,et al.Role of smoothmuscle cells in vascular calcification:implications in atherosclerosis andarterial stiffness[J].Cardiovascular Research,2018,114(4):590-600.[7]BOSTRÖM K I,RAJAMANNAN N M,TOWLER D A.The regulationof valvular and vascular sclerosis by osteogenic morphogens[J].Circulation Research,2011,109(5):564-577.[8]SPEER M Y,YANG H Y,BRABB T,et al.Smooth muscle cells giverise to osteochondrogenic precursors and chondrocytes incalcifying arteries[J].Circulation Research,2009,104(6):733-741.[9]PROUDFOOT D,SKEPPER J N,HEGYI L,et al.Apoptosisregulates human vascular calcification in vitro:evidence forinitiation of vascular calcification by apoptotic bodies[J].Circulation Research,2000,87(11):1055-1062.[10]AN S J,BOYD R,ZHU M,et al.NADPH oxidase mediatesangiotensin II-induced endothelin-1expression in vascularadventitial fibroblasts[J].Cardiovascular Research,2007,75(4):702-709.[11]ZEADIN M,BUTCHER M,WERSTUCK G,et al.Effect of leptin onvascular calcification in apolipoprotein E-deficient mice[J].Arterioscler Thromb Vasc Biol,2009,29(12):2069-2075. [12]LEOPOLD J A.Vascular calcification:mechanisms of vascularsmooth muscle cell calcification[J].Trends in CardiovascularMedicine,2015,25(4):267-274.[13]刘聿秀.高尿酸诱导血管钙化的机制研究[D].青岛:青岛大学,2015.[14]LIU Q,LUO Y,ZHAO Y,et al.Nano-hydroxyapatite acceleratesvascular calcification via lysosome impairment and autophagydysfunction in smooth muscle cells[J].Bioact Mater,2022,8:478-493.[15]LIANG J,HUANG J,HE W,et al.β-Hydroxybutyric Inhibits vascularcalcification via autophagy enhancement in models induced byhigh phosphate[J].Front Cardiovasc Med,2021,8:685748. [16]CICERI P,ELLI F,CAPPELLETTI L,et al.A new in vitro model todelay high phosphate-induced vascular calcification progression[J].Mol Cell Biochem,2015,410(1/2):197-206.[17]BYON C H,JAVED A,DAI Q,et al.Oxidative stress inducesvascular calcification through modulation of the osteogenictranscription factor Runx2by AKT signaling[J].The Journal ofBiological Chemistry,2008,283(22):15319-15327.[18]OUIMET M,FRANKLIN V,MAK E,et al.Autophagy regulatescholesterol efflux from macrophage foam cells via lysosomal acidlipase[J].Cell Metabolism,2011,13(6):655-667.[19]REYNOLDS J L,JOANNIDES A J,SKEPPER J N,et al.Humanvascular smooth muscle cells undergo vesicle-mediatedcalcification in response to changes in extracellular calcium andphosphate concentrations:a potential mechanism for acceleratedvascular calcification in ESRD[J].Journal of the AmericanSociety of Nephrology,2004,15(11):2857-2867.(收稿日期:2022-03-30)(本文编辑王雅洁)。
CellTiter Glo Luminescent Cell Viability Assay Protocol
Promega Corporation ·2800 Woods Hollow Road ·Madison, WI 53711-5399 USA Toll F ree in USA 800-356-9526·Phone 608-274-4330 ·F ax 608-277-2516 ·1.Description (1)2.Product Components and Storage Conditions (4)3.Performing the CellTiter-Glo ®Assay (5)A.Reagent Preparation (5)B.Protocol for the Cell Viability Assay (6)C.Protocol for Generating an ATP Standard Curve (optional) (7)4.Appendix (7)A.Overview of the CellTiter-Glo ®Assay..............................................................7B.Additional Considerations..................................................................................8C.References............................................................................................................11D.Related Products. (12)1.DescriptionThe CellTiter-Glo ®Luminescent Cell Viability Assay (a–e)is a homogeneous method to determine the number of viable cells in culture based on quantitation of the ATP present, which signals the presence of metabolically active cells. The CellTiter-Glo ®Assay is designed for use with multiwell-plate formats, making it ideal for automated high-throughput screening (HTS) and cell proliferation and cytotoxicity assays. The homogeneous assay procedure (Figure 1) involves adding a single reagent (CellTiter-Glo ®Reagent) directly to cells cultured in serum-supplemented medium. Cell washing, removal of medium or multiple pipetting steps are not required.The homogeneous “add-mix-measure” format results in cell lysis and generation of a luminescent signal proportional to the amount of ATP present (Figure 2).The amount of ATP is directly proportional to the number of cells present in culture in agreement with previous reports (1). The CellTiter-Glo ®Assay relies on the properties of a proprietary thermostable luciferase (Ultra-Glo™ Recombinant Luciferase), which generates a stable “glow-type” luminescent signal and improves performance across a wide range of assay conditions. The luciferase reaction for this assay is shown in Figure 3. The half-life of the luminescent signal resulting from this reaction is greater than five hours (Figure 4). This extended half-life eliminates the need for reagent injectors and provides flexibility for continuous or batch-mode processing of multiple plates. The unique homogeneous format reduces pipetting errors that may be introduced during the multiple steps required by other ATP-measurement methods.CellTiter-Glo ®Luminescent Cell Viability AssayAll technical literature is available on the Internet at: /protocols/ Please visit the web site to verify that you are using the most current version of this Technical Bulletin. Please contact Promega Technical Services if you have questions on useofthissystem.E-mail:********************Figure 1. Flow diagram showing preparation and use of CellTiter-Glo ®Reagent.Promega Corporation ·2800 Woods Hollow Road ·Madison, WI 53711-5399 USA Toll F ree in USA 800-356-9526·Phone 608-274-4330 ·F ax 608-277-2516 ·3170M A 12_0ACellTiter-Glo CellTiter-Glo MixerLuminometer®System Advantages•Homogeneous:“Add-mix-measure” format reduces the number of plate-handling steps to fewer than that required for similar ATP assays.•Fast:Data can be recorded 10 minutes after adding reagent.•Sensitive:Measures cells at numbers below the detection limits of standard colorimetric and fluorometric assays.•Flexible:Can be used with various multiwell formats. Data can be recorded by luminometer or CCD camera or imaging device.•Robust:Luminescent signal is very stable, with a half-life >5 hours,depending on cell type and culture medium used.•Able to Multiplex:Can be used with reporter gene assays or other cell-based assays from Promega (2,3).Figure 3. The luciferase reaction.Mono-oxygenation of luciferin is catalyzed byluciferase in the presence of Mg 2+, ATP and molecular oxygen.Promega Corporation ·2800 Woods Hollow Road ·Madison, WI 53711-5399 USA Toll F ree in USA 800-356-9526·Phone 608-274-4330 ·F ax 608-277-2516 ·3171M A 12_0A L u m i n e s c e n c e (R L U )Cells per Well10,00060,00020,00030,00040,00050,0000R² = 0.9990.5 × 1061.0 × 1061.5 × 1062.0 × 1062.5 × 1063.0 × 1063.5 × 1064.0 × 106r² = 0.99020,00010,00030,00040,00050,000r² = 0.9900100200300400HO SN S N O S N S N OCOOH +ATP+O 2Ultra-Glo™ Recombinant Luciferase +AMP+PP i +CO 2+LightBeetle Luciferin OxyluciferinMg 2+0Figure 2. Cell number correlates with luminescent output.A direct relationship exists between luminescence measured with the CellTiter-Glo ®Assay and the number of cells in culture over three orders of magnitude. Serial twofold dilutions of HEK293cells were made in a 96-well plate in DMEM with 10% FBS, and assays wereperformed as described in Section 3.B. Luminescence was recorded 10minutes after reagent addition using a GloMax ®-Multi+ Detection System. Values represent the mean ± S.D. of four replicates for each cell number. The luminescent signal from 50HEK293 cells is greater than three times the background signal from serum-supplemented medium without cells. There is a linear relationship (r 2= 0.99)between the luminescent signal and the number of cells from 0to 50,000 cells per well.Figure 4. Extended luminescent half-life allows high-throughput batchprocessing.Signal stability is shown for three common cell lines. HepG2 and BHK-21cells were grown and assayed in MEM containing 10% FBS, while CHO-K1 cells were grown and assayed in DME/F-12 containing 10% FBS. CHO-K1, BHK-21 and HepG2 cells, at 25,000 cells per well, were added to a 96-well plate. After an equal volume of CellTiter-Glo ®Reagent was added, plates were shaken and luminescence monitored over time with the plates held at 22°C. The half-lives of the luminescent signals for the CHO-K1, BHK-21 and HepG2 cells were approximately 5.4, 5.2 and5.8hours, respectively.2.Product Components and Storage ConditionsProduct Size Cat.#CellTiter-Glo ®Luminescent Cell Viability Assay 10ml G7570Substrate is sufficient for 100 assays at 100µl/assay in 96-well plates or 400 assays at 25µl/assay in 384-well plates. Includes:• 1 × 10mlCellTiter-Glo ®Buffer • 1 vial CellTiter-Glo ®Substrate (lyophilized)Product Size Cat.#CellTiter-Glo ®Luminescent Cell Viability Assay 10 × 10ml G7571Each vial of substrate is sufficient for 100 assays at 100µl/assay in 96-well plates or 400 assays at 25µl/assay in 384-well plates (1,000 to 4,000 total assays). Includes:•10 × 10mlCellTiter-Glo ®Buffer •10 vials CellTiter-Glo ®Substrate (lyophilized)Promega Corporation ·2800 Woods Hollow Road ·Madison, WI 53711-5399 USA Toll F ree in USA 800-356-9526·Phone 608-274-4330 ·F ax 608-277-2516 ·R e l a t i v e L u m i n e s c e n c e (%)Time (minutes)CHO-K101020304050607080901003173M A 12_0AProduct Size Cat.# CellTiter-Glo®Luminescent Cell Viability Assay100ml G7572 Substrate is sufficient for 1,000 assays at 100µl/assay in 96-well plates or 4,000assays at 25µl/assay in 384-well plates. Includes:•1 × 100ml CellTiter-Glo®Buffer• 1 vial CellTiter-Glo®Substrate (lyophilized)Product Size Cat.# CellTiter-Glo®Luminescent Cell Viability Assay10 × 100ml G7573Each vial of substrate is sufficient for 1,000 assays at 100µl/assay in 96-well plates or4,000 assays at 25µl/assay in 384-well plates (10,000to 40,000 total assays). Includes:•10 × 100ml CellTiter-Glo®Buffer•10 vials CellTiter-Glo®Substrate (lyophilized)Storage Conditions:For long-term storage, store the lyophilized CellTiter-Glo®Substrate and CellTiter-Glo®Buffer at –20°C. For frequent use, the CellTiter-Glo®Buffer can be stored at 4°C or room temperature for 48hours without loss of activity. See product label for expiration date information. ReconstitutedCellTiter-Glo®Reagent (Buffer plus Substrate) can be stored at room temperaturefor up to 8hours with <10% loss of activity, at 4°C for 48hours with ~5% lossof activity, at 4°C for 4days with ~20% loss of activity or at –20°C for 21weekswith ~3% loss of activity. The reagent is stable for up to ten freeze-thaw cycles,with less than 10% loss of activity.3.Performing the CellTiter-Glo®AssayMaterials to Be Supplied by the User•opaque-walled multiwell plates adequate for cell culture•multichannel pipette or automated pipetting station for reagent delivery•device (plate shaker) for mixing multiwell plates•luminometer, CCD camera or imaging device capable of reading multiwell plates •optional:ATP for use in generating a standard curve (Section 3.C)3.A.Reagent Preparation1.Thaw the CellTiter-Glo®Buffer, and equilibrate to room temperature priorto use. For convenience the CellTiter-Glo®Buffer may be thawed andstored at room temperature for up to 48hours prior to use.2.Equilibrate the lyophilized CellTiter-Glo®Substrate to room temperatureprior to use.Promega Corporation·2800 Woods Hollow Road ·Madison, WI 53711-5399 USA Toll F ree in USA 800-356-9526·Phone 608-274-4330 ·F ax 608-277-2516 ·3.A.Reagent Preparation (continued)3.Transfer the appropriate volume (10ml for Cat.# G7570 and G7571, or 100mlfor Cat.# G7572 and G7573) of CellTiter-Glo ®Buffer into the amber bottlecontaining CellTiter-Glo ®Substrate to reconstitute the lyophilizedenzyme/substrate mixture. This forms the CellTiter-Glo ®Reagent.4.Mix by gently vortexing, swirling or inverting the contents to obtain ahomogeneous solution. The CellTiter-Glo ®Substrate should go intosolution easily in less than 1minute.3.B.Protocol for the Cell Viability AssayWe recommend that you perform a titration of your particular cells todetermine the optimal number and ensure that you are working within thelinear range of the CellTiter-Glo ®Assay. Figure 2 provides an example of sucha titration of HEK293 cells using 0 to 50,000 cells per well in a 96-well format.1.Prepare opaque-walled multiwell plates with mammalian cells in culturemedium, 100µl per well for 96-well plates or 25µl per well for 384-wellplates.Multiwell plates must be compatible with the luminometer used.2.Prepare control wells containing medium without cells to obtain a value forbackground luminescence.3.Add the test compound to experimental wells, and incubate according toculture protocol.4.Equilibrate the plate and its contents at room temperature forapproximately 30 minutes.5.Add a volume of CellTiter-Glo ®Reagent equal to the volume of cell culturemedium present in each well (e.g., add 100µl of reagent to 100µl of mediumcontaining cells for a 96-well plate, or add 25µl of reagent to 25µl ofmedium containing cells for a 384-well plate).6.Mix contents for 2 minutes on an orbital shaker to induce cell lysis.7.Allow the plate to incubate at room temperature for 10 minutes to stabilizeluminescent signal.Note:Uneven luminescent signal within standard plates can be caused bytemperature gradients, uneven seeding of cells or edge effects in multiwellplates.8.Record luminescence.Note:Instrument settings depend on the manufacturer. An integration timeof 0.25–1 second per well should serve as a guideline.Promega Corporation ·2800 Woods Hollow Road ·Madison, WI 53711-5399 USA Toll F ree in USA 800-356-9526·Phone 608-274-4330 ·F ax 608-277-2516 ·3.C.Protocol for Generating an ATP Standard Curve (optional)It is a good practice to generate a standard curve using the same plate onwhich samples are assayed. We recommend ATP disodium salt (Cat.# P1132,Sigma Cat.# A7699 or GE Healthcare Cat.# 27-1006). The ATP standard curveshould be generated immediately prior to adding the CellTiter-Glo®Reagentbecause endogenous ATPase enzymes found in sera may reduce ATP levels.1.Prepare 1µM ATP in culture medium (100µl of 1µM ATP solution contains10–10moles ATP).2.Prepare serial tenfold dilutions of ATP in culture medium (1µM to 10nM;100µl contains 10–10to 10–12moles of ATP).3.Prepare a multiwell plate with varying concentrations of ATP standard in100µl medium (25µl for a 384-well plate).4.Add a volume of CellTiter-Glo®Reagent equal to the volume of ATPstandard present in each well.5.Mix contents for 2 minutes on an orbital shaker.6.Allow the plate to incubate at room temperature for 10 minutes to stabilizethe luminescent signal.7.Record luminescence.4.Appendix4.A.Overview of the CellTiter-Glo®AssayThe assay system uses the properties of a proprietary thermostable luciferase toenable reaction conditions that generate a stable “glow-type” luminescentsignal while simultaneously inhibiting endogenous enzymes released duringcell lysis (e.g., ATPases). Release of ATPases will interfere with accurate ATPmeasurement. Historically, firefly luciferase purified from Photinus pyralis(LucPpy) has been used in reagents for ATP assays (1,4–7). However, it hasonly moderate stability in vitro and is sensitive to its chemical environment,including factors such as pH and detergents, limiting its usefulness fordeveloping a robust homogeneous ATP assay. Promega has successfullydeveloped a stable form of luciferase based on the gene from another firefly,Photuris pennsylvanica(LucPpe2), using an approach to select characteristics thatimprove performance in ATP assays. The unique characteristics of this mutant(LucPpe2m) enabled design of a homogeneous single-reagent-addition approachto perform ATP assays with cultured cells. Properties of the CellTiter-Glo®Reagent overcome the problems caused by factors, such as ATPases, thatinterfere with ATP measurement in cell extracts. The reagent is physicallyrobust and provides a sensitive and stable luminescent output.Promega Corporation·2800 Woods Hollow Road ·Madison, WI 53711-5399 USA Toll F ree in USA 800-356-9526·Phone 608-274-4330 ·F ax 608-277-2516 ·4.A.Overview of the CellTiter-Glo®Assay (continued)Sensitivity and Linearity:The ATP-based detection of cells is more sensitivethan other methods (8–10). In experiments performed by Promega scientists,the luminescent signal from 50HEK293 cells is greater than three standarddeviations above the background signal from serum-supplemented mediumwithout cells. There is a linear relationship (r2= 0.99) between the luminescentsignal and the number of cells from 0 to 50,000 cells per well in the 96-wellformat. The luminescence values in Figure 2 were recorded after 10minutes ofincubation at room temperature to stabilize the luminescent signal as describedin Section3.B. Incubation of the same 96-well plate used in the experimentshown in Figure 2 for 360minutes at room temperature had little effect on therelationship between luminescent signal and number of cells (r2= 0.99).Speed:The homogeneous procedure to measure ATP using the CellTiter-Glo®Assay is quicker than other ATP assay methods that require multiple steps toextract ATP and measure luminescence. The CellTiter-Glo®Assay also is fasterthan other commonly used methods to measure the number of viable cells(such as MTT, alamarBlue®or Calcein-AM) that require prolonged incubationsteps to enable the cells’ metabolic machinery to convert indicator moleculesinto a detectable signal.4.B.Additional ConsiderationsTemperature:The intensity and decay rate of the luminescent signal from theCellTiter-Glo®Assay depends on the luciferase reaction rate. Environmentalfactors that affect the luciferase reaction rate will change the intensity andstability of the luminescent signal. Temperature is one factor that affects therate of this enzymatic assay and thus the light output. For consistent results,equilibrate assay plates to a constant temperature before performing the assay.Transferring eukaryotic cells from 37°C to room temperature has little effect onATP content (5). We have demonstrated that removing cultured cells from a37°C incubator and allowing them to equilibrate to 22°C for 1–2 hours hadlittle effect on ATP content. For batch-mode processing of multiple assayplates, take precautions to ensure complete temperature equilibration. Platesremoved from a 37°C incubator and placed in tall stacks at room temperaturewill require longer equilibration than plates arranged in a single layer.Insufficient equilibration may result in a temperature gradient effect betweenwells in the center and at the edge of the plates. The temperature gradientpattern also may depend on the position of the plate in the stack.Promega Corporation·2800 Woods Hollow Road ·Madison, WI 53711-5399 USA Toll F ree in USA 800-356-9526·Phone 608-274-4330 ·F ax 608-277-2516 ·Chemicals:The chemical environment of the luciferase reaction affects theenzymatic rate and thus luminescence intensity. Differences in luminescenceintensity have been observed using different types of culture media and sera.The presence of phenol red in culture medium should have little impact onluminescence output. Assaying 0.1µM ATP in RPMI medium without phenolred resulted in ~5% increase in luminescence output (in relative light units[RLU]) compared to assays in RPMI containing the standard concentration ofphenol red, whereas assays in RPMI medium containing twice the normalconcentration of phenol red showed a ~2% decrease in luminescence.Solvents for the various test compounds may interfere with the luciferasereaction and thus the light output from the assay. Interference with theluciferase reaction can be detected by assaying a parallel set of control wellscontaining medium without cells. Dimethylsulfoxide (DMSO), commonly usedas a vehicle to solubilize organic chemicals, has been tested at finalconcentrations of up to 2% in the assay and only minimally affects light output.Plate Recommendations:We recommend using standard opaque-walledmultiwell plates suitable for luminescence measurements. Opaque-walledplates with clear bottoms to allow microscopic visualization of cells also maybe used; however, these plates will have diminished signal intensity andgreater cross talk between wells. Opaque white tape may be used to decreaseluminescence loss and cross talk.Cellular ATP Content:Different cell types have different amounts of ATP,and values reported for the ATP level in cells vary considerably (1,4,11–13).Factors that affect the ATP content of cells may affect the relationship betweencell number and luminescence. Anchorage-dependent cells that undergocontact inhibition at high densities may show a change in ATP content per cellat high densities, resulting in a nonlinear relationship between cell numberand luminescence. Factors that affect the cytoplasmic volume or physiology ofcells also will affect ATP content. For example, oxygen depletion is one factorknown to cause a rapid decrease in ATP (1).Promega Corporation·2800 Woods Hollow Road ·Madison, WI 53711-5399 USA Toll F ree in USA 800-356-9526·Phone 608-274-4330 ·F ax 608-277-2516 ·4.B.Additional Considerations (continued)Mixing:Optimal assay performance is achieved when the CellTiter-Glo®Reagent is mixed completely with the cultured cells. Suspension cell lines (e.g., Jurkat cells) generally require less mixing to achieve lysis and extract ATP than adherent cells (e.g., L929 cells). Tests were done to evaluate the effect ofshaking the plate after adding the CellTiter-Glo® Reagent. Suspension cellscultured in multiwell plates showed only minor differences in light outputwhether or not the plates were shaken after adding the CellTiter-Glo®Reagent.Adherent cells are more difficult to lyse and show a substantial differencebetween shaken and nonshaken plates.Several additional parameters related to reagent mixing include the force ofdelivery of CellTiter-Glo®Reagent, sample volume and dimensions of the well.All of these factors may affect assay performance. The degree of reagent mixing required may be affected by the method used to add the CellTiter-Glo®Reagent to the assay plates. Automated pipetting devices using a greater or lesser force of fluid delivery may affect the degree of subsequent mixing required.Complete reagent mixing in 96-well plates should be achieved using orbitalplate shaking devices built into many luminometers and the recommended2-minute shaking time. Special electromagnetic shaking devices that use aradius smaller than the well diameter may be required to efficiently mixcontents of 384-well plates. The depth of medium and geometry of themultiwell plates may have an effect on mixing efficiency. We recommend that you take these factors into consideration when performing the assay andempirically determine whether a mixing step is necessary for the individualapplication.LuminometersFor highly sensitive luminometric assays, the luminometer model and settings greatly affect the quality of data obtained. Luminometers from differentmanufacturers will vary in sensitivities and dynamic ranges. We recommend the GloMax®products because these instruments do not require gainadjustments to achieve optimal sensitivity and dynamic range. Additionally, GloMax®instruments are preloaded with Promega protocols for ease of use.If you are not using a GloMax®luminometer, consult the operating manual for your luminometer to determine the optimal settings. The limits should beverified on each instrument before analysis of experimental samples. The assay should be linear in some portion of the detection range of the instrument used.For an individual luminometer there may be different gain settings. Werecommend that you optimize the gain settings.4.C.References1.Crouch, S.P. et al.(1993) The use of ATP bioluminescence as a measure of cellproliferation and cytotoxicity. J. Immunol. Methods160, 81–8.2.Farfan, A.et al.(2004) Multiplexing homogeneous cell-based assays. Cell Notes10, 2–5.3.Riss, T., Moravec, R. and Niles, A. (2005) Selecting cell-based assays for drugdiscovery screening. Cell Notes13, 16–21.4.Kangas, L., Grönroos, M. and Nieminen, A.L. (1984) Bioluminescence of cellular ATP:A new method for evaluating cytotoxic agents in vitro. Med. Biol.62, 338–43.5.Lundin, A. et al.(1986) Estimation of biomass in growing cell lines by adenosinetriphosphate assay.Methods Enzymol. 133, 27–42.6.Sevin, B.U. et al.(1988) Application of an ATP-bioluminescence assay in human tumorchemosensitivity testing. Gynecol. Oncol.31, 191–204.7.Gerhardt, R.T.et al.(1991) Characterization of in vitro chemosensitivity ofperioperative human ovarian malignancies by adenosine triphosphatechemosensitivity assay. Am. J. Obstet. Gynecol. 165, 245–55.8.Petty, R.D. et al.(1995) Comparison of MTT and ATP-based assays for themeasurement of viable cell number. J. Biolumin. Chemilumin.10, 29–34.9.Cree, I.A. et al.(1995) Methotrexate chemosensitivity by ATP luminescence in humanleukemia cell lines and in breast cancer primary cultures: Comparison of the TCA-100assay with a clonogenic assay. AntiCancer Drugs6, 398–404.10.Maehara, Y. et al.(1987) The ATP assay is more sensitive than the succinatedehydrogenase inhibition test for predicting cell viability. Eur. J. Cancer Clin. Oncol.23, 273–6.11.Stanley, P.E. (1986) Extraction of adenosine triphosphate from microbial and somaticcells. Methods Enzymol.133, 14–22.12.Beckers, B. et al.(1986) Application of intracellular ATP determination in lymphocytesfor HLA-typing. J. Biolumin. Chemilumin.1, 47–51.13.Andreotti, P.E. et al.(1995) Chemosensitivity testing of human tumors using amicroplate adenosine triphosphate luminescence assay: Clinical correlation forcisplatin resistance of ovarian carcinoma. Cancer Res. 55, 5276–82.4.D.Related ProductsCell Proliferation ProductsProduct Size Cat.# ApoLive-Glo™ Multiplex Assay10ml G6410 ApoTox-Glo™ Triplex Assay10ml G6320 CellTiter-Fluor™ Cell Viability Assay (fluorescent)10ml G6080 CellTiter-Blue®Cell Viability Assay (resazurin)20ml G8080 CellTiter 96®AQ ueous One SolutionCell Proliferation Assay (MTS, colorimetric)200 assays G3582 CellTiter 96®AQ ueous Non-RadioactiveCell Proliferation Assay (MTS, colorimetric)1,000 assays G5421 CellTiter 96®AQ ueous MTS Reagent Powder1g G1111 CellTiter 96®Non-RadioactiveCell Proliferation Assay (MTT, colorimetric)1,000 assays G4000 Additional sizes available.Cytotoxicity AssaysProduct Size Cat.# CytoTox-Glo™ Cytotoxicity Assay (luminescent)*10ml G9290Mitochondrial ToxGlo™ Assay*10ml G8000 MultiTox-Glo Multiplex Cytotoxicity Assay(luminescent, fluorescent)*10ml G9270 MultiTox-Fluor Multiplex Cytotoxicity Assay(fluorescent)*10ml G9200 CytoTox-Fluor™ Cytotoxicity Assay (fluorescent)*10ml G9260 CytoTox-ONE™ Homogeneous MembraneIntegrity Assay (LDH, fluorometric)*200–800 assays G7890 CytoTox-ONE™ Homogeneous MembraneIntegrity Assay, HTP1,000–4,000 assays G7892 CytoTox 96® Non-Radioactive Cytotoxicity Assay1,000 assays G1780 (LDH, colorimetric)*GSH-Glo™ Glutathione Assay10ml V691150ml V6912 GSH/GSSG-Glo™ Assay10ml V661150ml V6612 *Additional sizes available.LuminometersProduct Size Cat.# GloMax®-Multi+ Detection System with Instinct™ Software:Base Instrument with Shaking 1 each E8032 GloMax®-Multi+ Detection System with Instinct™ Software:Base Instrument with Heating and Shaking 1 each E9032 GloMax®-Multi+ Luminescence Module 1 each E8041Apoptosis ProductsProduct Size Cat.# Caspase-Glo®2 Assay*10ml G0940 Caspase-Glo®6 Assay*10ml G0970 Caspase-Glo®3/7 Assay* 2.5ml G8090 Caspase-Glo®8 Assay* 2.5ml G8200 Caspase-Glo®9 Assay* 2.5ml G8210Apo-ONE®Homogeneous Caspase-3/7 Assay1ml G7792 DeadEnd™ Fluorometric TUNEL System60 reactions G3250 DeadEnd™ Colorimetric TUNEL System20 reactions G7360Anti-ACTIVE®Caspase-3 pAb50µl G7481Anti-PARP p85 Fragment pAb50µl G7341Anti-pS473Akt pAb40µl G7441 Caspase Inhibitor Z-VAD-FMK, 20mM50µl G7231125µl G7232*Additional sizes available.(a)U.S. Pat. Nos. 6,602,677 and 7,241,584, European Pat. No. 1131441, Japanese Pat. Nos. 4537573 and 4520084 and other patents pending(b)U.S. Pat. No. 7,741,067, Japanese Pat. No. 4485470 and other patents pending.(c)U.S. Pat. No. 7,700,310, European Pat. No. 1546374 and other patents pending.(d)U.S. Pat. Nos 7,083,911, 7,452,663 and 7,732,128, European Pat. No. 1383914 and Japanese Pat. Nos. 4125600 and 4275715.(e)The method of recombinant expression of Coleoptera luciferase is covered by U.S. Pat. Nos. 5,583,024, 5,674,713 and 5,700,673.© 2001–2012 Promega Corporation. All Rights Reserved.Anti-ACTIVE, Apo-ONE, Caspase-Glo, CellTiter 96, CellTiter-Blue, CellTiter-Glo, CytoTox 96 and GloMax are registered trademarks of Promega Corporation. ApoTox-Glo, ApoLive-Glo, CellTiter-Fluor, CytoTox-Fluor, CytoTox-Glo, CytoTox-ONE, DeadEnd, GSH-Glo, GSH/GSSG-Glo, Instinct, Mitochondrial ToxGlo and Ultra-Glo are trademarks of Promega Corporation. alamarBlue is a registered trademark of Trek Diagnostic Ssystems, Inc.Products may be covered by pending or issued patents or may have certain limitations. Please visit our Web site for more information.All prices and specifications are subject to change without prior notice.Product claims are subject to change. Please contact Promega Technical Services or access the Promega online catalog for the most up-to-date information on Promega products.。
美洲大蠊抗肝纤维化活性提取物ML-D中粘多糖的含量测定
ｆ ｒ ｏ ｍ Ｐ ｅ ｒ ｉ ｐｌ ａ ｎ ｅ ｔ ａ ａ ｍｅ ｒ ｉ ｃ ａ ｉ ｒ ａ ．Ｍ ｅ ｔ ｈ ｏ ｄ ｓ ：Ｐ ｈ ｅ ｎ ｏ ｌ — ｖ ｉ ｔ ｉ ｒ ｏ ｌ ｃ ｏ ｌ ｏ ｒ ｉ ｍｅ ｔ ｙ ｒ ｗａ ｓ ｕ ｓ ｅ ｄ ｆ ｏ ｒ ｄ ｅ ｔ ｅ ｍｉ ｒ ｎ ｉ ｎ ｇ ｃ ｏ ｎ ｔ ｅ ｎ ｔ ｏ ｆ ｙ ｃ ｏ ｓ ａ ｍｉ ｎ ｏ ｙ ｃ ａ ｎ ；ｏ ｒ ｔ ｈ ｏ ｇ ｏ ｎ ａ ｌ
美洲大蠊抗肝纤维化活性提取物Ｍ Ｌ — Ｄ 中粘多糖的含量测定
李夸巧 ， 李春艳 ， 杨 永寿 ， 陈丽娜 ， 肖培云
（ 大理 学院 药学与 化 学 学院 ， 云 南 大理 ６ ７ １ ０ ０ ０ ）
［ 摘要 ］目的 ： 建立美洲大蠊抗肝纤维化活性提 取物ＭＬ — Ｄ中粘 多糖的含量测定方法 。方法 ： 采用苯酚一 硫酸法测定粘多糖的含 量， 采用 正交试验设计考察ＭＬ — Ｄ中粘 多糖 的水解工 艺和 显色条件 ， 采用单 因素试验考察加热显色时间、 苯酚和硫酸的用量。结
ｄ ｅ ｓ ｉ ｇ ｎ ｋ（ ３ ４ ） ｗ ａ ｓ ａ ｄ ｏ ｐ ｔ ｅ ｄ ｔ ｏ ｉ ｎ ｖ ｅ ｓ ｔ ｉ ｇ ａ ｔ ｅ ｈ ｙ ｄ ｒ ｏ ｌ ｙ ｓ ｉ ｓ ｐ ｒ ｏ ｃ ｅ ｓ ｓ ｏ ｆ ｌ ｇ ｙ ｃ ｏ ｓ ａ ｍｉ ｎ ｏ ｇ ｌ ｙ ｃ ａ ｎ ｉ ｎ ＭＬ — Ｄ ； ｓ ｉ ｎ ｇ ｌ ｅ ｆ ａ ｃ ｔ ｏ ｒ ｅ ｘ ｐ ｅ ｉ ｒ ｍ ｅ ｎ ｔ ｗ ａ ｓ ｕ ｓ ｅ ｄ ｔ ｏ ｉ ｎ ｖ ｅ ｓ ｔ ｉ ｇ ａ ｔ ｅ
dammarane达玛烷-生物化学与分子生物学
Dactinomycin 放线菌素Ddammarane type 达玛烷型dammarane 达玛烷dampening of charge effects 电荷效应的衰减Dane particle 丹氏粒[乙型肝炎病毒的完整毒粒] dansyl 丹（磺）酰，1-二甲氨基萘-5-磺酰dansyl chloride 丹磺酰氯dansyl method 丹磺酰法dantrolene 硝苯呋海因[肌肉松弛剂]dapsone 氨苯砜dark adaptation 暗适应dark current 暗电流dark field microscope 暗视野显微镜，暗视场显微镜dark field microscopy 暗视野显微术，暗视场显微术dark field 暗视野，暗视场dark reaction 暗反应dark repair 暗修复dark respiration 暗呼吸dark room 暗室，暗房dark seed 需暗种子Darwinian selection 达尔文选择DAS 下游激活部位data accumulation 数据积累data acquisition 数据获取data analysis 数据分析data bank 数据库data base 数据库data handling 数据处理data logger 数据记录器data logging 数据记录data output 数据输出data processing 数据处理data recording 数据记录dative bond 配位键dative covalent bond 配位共价键dauermodification 持续饰变daughter cell 子代细胞daughter chromatid 子染色单体daughter chromosome 子染色体daughter colony 子菌落[由原生菌落续发生长的小菌落] daunomycin 道诺霉素daunorubicin 道诺红菌素day vision 昼视觉，昼视力de novo sequencing 从头测序de novo synthesis 从头合成deactivation 去活化（作用），失活（作用），钝化deacylated tRNA 脱酰tRNAdead time 死时间dead volume 死体积deadenylation 脱腺苷化deadhesion factor 去黏附因子DEAE Sephacel [商]DEAE-葡聚糖纤维素，二乙氨乙基葡聚糖纤维素DEAE-cellulose DEAE-纤维素DEAE-dextran DEAE-葡聚糖dealkylation 脱烷基化deaminase 脱氨酶deamination 脱氨（基）death ligand 死亡配体death phase 死亡期[如见于细胞生长曲线]death point 死点deblocking 去封闭debranching enzyme 脱支酶，支链淀粉酶debris 碎片，残渣decahedron 十面体decamer 十体decamethonium 十烷双胺，癸烷双胺decane 癸烷decanoates 癸酸decantation 倾析decanting 倾析decapacitation 去（获）能decapsidate 脱衣壳decarboxylase 脱羧酶decarboxylation 脱羧（作用）decatenation 解连环{作用}decay accelerating factor 衰变加速因子decay chain 衰变链decay constant 衰变常数decay 原因不明腐败deceleration phase 减速期[如见于细胞生长曲线]dechlorination 脱氯作用deciduous leaf 落叶decline phase [细胞生长曲线的]衰亡期decoagulant 抗凝剂decoding 译码，解码decoginine 德夸菌素decomposer 分解者[可指具有分解动植物残体或其排泄物能力的微生物]decompression 降压，减压decondensation 解凝（聚）decontaminant 净化剂，去污剂decontaminating agent 净化剂，去污剂decontamination 净化，去污decorin 核心蛋白聚糖[一种基质蛋白聚糖，又称为PG-40] decoy receptor 诱杀受体decoy 诱杀物dedifferentiation 去分化，脱分化deep colony 深层菌落deep etching 深度蚀刻deep jet fermentor 深部喷注发酵罐deep refrigeration 深度冷冻deep shaft system 深井系统[如用于污水处理] defasciculation factor 解束因子[取自水蛭，可破坏神经束] defective interfering particle 缺损干扰颗粒，干扰缺损颗粒defective interfering RNA 缺损干扰RNAdefective interfering virus 缺损干扰病毒defective interfering 缺损干扰defective mutant 缺损突变体，缺陷突变型，缺陷突变株defective phage 缺损噬菌体，缺陷噬菌体defective virus 缺损病毒，缺陷病毒defective 缺损的，缺陷的defense peptide 防卫肽defense response 防御反应，防卫反应defense 防御，防卫defensin 防卫素[动物细胞的内源性抗菌肽]deficiency 缺乏，缺损，缺陷deficient 缺少的，缺损的，缺陷的defined medium 确定成分培养基，已知成分培养液defined 确定的defintion 定义defoliating agent 脱叶剂defoliation 脱叶deformylase 去甲酰酶[见于原核细胞，作用于甲酰甲硫氨酸] degasser 脱气装置degassing 脱气，除气degeneracy 简并；简并性，简并度degenerate codon 简并密码子degenerate oligonucleotide 简并寡核苷酸degenerate primer 简并引物degenerate sequence 简并序列degenerate 简并的degeneration 退化，变性degenerin 退化蛋白[与某些感觉神经元的退化有关] deglycosylation 去糖基化degradable polymer 降解性高分子degradation 降解degranulation 脱（颗）粒（作用）degree of acidity 酸度degree of dominance 显性度degree of polymerization 聚合度degron 降解决定子[决定某一蛋白发生降解或部分降解的序列要素] deguelin 鱼藤素dehalogenation 脱卤（作用）dehardening 解除锻炼dehumidifier 除湿器dehydratase 脱水酶dehydrated medium 干燥培养基dehydration 脱水（作用）dehydroepiandrosterone 脱氢表雄酮dehydrogenase 脱氢酶dehydrogenation 脱氢（作用）dehydroluciferin 脱氢萤光素deionization 去离子（作用）deionized water 去离子水deionized 去离子的deionizing 去离子（处理）delayed early transcription （延）迟早期转录[可特指病毒]delayed fluorescence 延迟荧光delayed heat 延迟热delayed hypersensitivity 延迟（型）超敏反应delayed ingeritance 延迟遗传delayed type hypersensitivity 迟发型超敏反应deletant 缺失体deletion mapping 缺失定位，缺失作图deletion mutagenesis 缺失诱变deletion mutant 缺失突变体deletion mutantion 缺失突变deletion 缺失deletional recombination 缺失重组delignification 脱木质化（作用）deliquescence 潮解delivery flask 分液瓶delocalized bond 离域键deltorphin delta啡肽[见于叶泡蛙皮肤，对delta阿片样肽受体的亲和力和选择性很强]demasking 解蔽demethylation 脱甲基化demineralized water 脱矿质水demulsifier 破乳剂demyelination 脱髓鞘denaturant gel 变性凝胶denaturant 变性剂denaturation 变性（作用）denatured DNA 变性DNAdenatured protein 变性蛋白（质）denatured 变性的denaturing gel electrophoresis 变性凝胶电泳denaturing gel 变性凝胶denaturing gradient polyacrylamide gel 变性梯度聚丙烯酰胺凝胶denaturing polyacrylamide gel 变性聚丙烯酰胺凝胶dendrite 树突dendritic cell 树状细胞dendrotoxin 树眼睛蛇毒素dengue virus 登革病毒denitrification 反硝化作用denitrifying bacteria 反硝化细菌denitrogen 排氮densimeter 密度计densitometer 光密度计densitometry 光密度（测定）法density gradient centrifugation 密度梯度离心density gradient electrophoresis 密度梯度电泳density gradient 密度梯度density 密度densovirus 浓核病毒dentritic cell 树枝状细胞Denver system [染色体组型]丹佛体制deoxyadenosine moniphosphate 脱氧腺苷（一磷）酸deoxyadenosine triphosphate 脱氧腺苷三磷酸deoxyadenosine 脱氧腺苷deoxyadenylate 脱氧腺苷酸deoxyadenylic acid 脱氧腺苷酸deoxycholate 脱氧胆酸盐deoxycorticosterone 脱氧皮质酮deoxycytidine monophosphate 脱氧胞苷（一磷）酸deoxycytidine triphosphate 脱氧胞苷三磷酸deoxycytidine 脱氧胞苷deoxycytidylate 脱氧胞苷酸deoxycytidylic acid 脱氧胞苷酸deoxyformycin 脱氧型霉素deoxygenation 脱氧deoxyglucose 脱氧葡萄糖deoxyguanosine monophosphate 脱氧鸟苷（一磷）酸deoxyguanosine triphosphate 脱氧鸟苷三磷酸deoxyguanosine 脱氧鸟苷deoxyguanylate 脱氧鸟苷酸deoxyguanylic acid 脱氧鸟苷酸deoxyhemoglobin 脱氧血红蛋白，去氧血红蛋白deoxyinosine triphosphate 脱氧肌苷三磷酸，脱氧次黄苷三磷酸deoxyinosine 脱氧肌苷，脱氧次黄苷deoxynivalenol 脱氧萎镰菌醇deoxynucleoside methylphosphonate 脱氧核苷膦酸甲酯deoxynucleoside phosphoramidite 脱氧核苷亚磷酰胺deoxynucleoside 脱氧核苷deoxynucleotide 脱氧核苷酸deoxyriboaldolase 脱氧核糖醛缩酶deoxyribomutase 脱氧核糖变位酶deoxyribonuclease 脱氧核糖核酸酶，DNA酶[包括DNA酶I、DNA酶II等] deoxyribonucleic acid 脱氧核糖核酸deoxyribonucleoside diphosphate 脱氧（核糖）核苷二磷酸deoxyribonucleoside hydrogenphosphonate 脱氧（核糖）核苷氢膦酸酯deoxyribonucleoside methylphosphonate 脱氧（核糖）核苷膦酸甲酯deoxyribonucleoside monophosphate 脱氧（核糖）核苷（一磷）酸deoxyribonucleoside phosphoramidite 脱氧（核糖）核苷亚磷酰胺deoxyribonucleoside triphosphate 脱氧（核糖）核苷三磷酸deoxyribonucleoside 脱氧（核糖）核苷deoxyribonucleotide 脱氧（核糖）核苷酸deoxyribose 脱氧核糖deoxysugar 脱氧糖deoxythymidine monophosphate 脱氧胸苷（一磷）酸deoxythymidine triphosphate 脱氧胸苷三磷酸deoxythymidine 脱氧胸腺嘧啶核苷，脱氧胸苷deoxythymidylate 脱氧胸苷酸deoxythymidylic acid 脱氧胸苷酸deoxyuridine monophosphate 脱氧尿苷（一磷）酸deoxyuridine 脱氧尿苷deoxyuridylate 脱氧尿苷酸deoxyuridylic acid 脱氧尿苷酸depactin [肌动蛋白]蚕食蛋白dependovirus 依赖病毒（属）[模式成员是腺伴随病毒] dephosphorylation 去磷酸化（作用）deplasmolysis 质壁分离复原depletion 耗竭，除尽depolarization 去极化，脱极化；解偏振（作用）depollution 去污染depolymerase 解聚酶depolymerization 解聚deposit 沉积；沉积物deposition 沉积（作用）；沉积物depressed 脱阻抑的depression 脱阻抑；压抑deprivation 除去，丧失deprived 除去的，丧失的depropagation 负增长反应，逆增长反应deprotection 脱保护deprotonation 去质子化，脱质子化depurination 脱嘌呤（作用）derepressed 脱阻抑的derepression 脱阻抑derivative 衍生物derivatization 衍生化（作用）derivave spectrum 导数光谱dermatan sulfate 硫酸皮肤素dermatogen 表皮原dermatoglyphics 肤纹学dermatonecrotoxin 皮肤坏死毒素dermatophagoides 嗜皮螨属dermenkaphaline 皮脑啡肽dermis 真皮dermorphin 皮啡肽desalting 脱盐desaminase 脱氨酶desaturase 脱饱和酶，脱氢酶desaturation 脱饱和，去饱和descending development 下行展开（法）descending facilitatory system 下行易化系统[用于神经生物学]descending inhibition 下行抑制descending inhibitory system 下行抑制系统desensitization 脱敏（作用）desetope (决定簇）选择位[抗原呈递中，II类主要组织相容性复合体与抗原相互作用的部位] desiccant 干燥剂desiccation 干燥（作用）desiccator cabinet 干燥橱desiccator 干燥器，保干器desmin 结蛋白desmocalmin 桥粒钙调（蛋白）结合蛋白，桥粒钙蛋白desmocollin 桥粒（芯）胶（粘）蛋白[见于桥粒芯的胶粘层，是桥粒芯蛋白的剪接变体] desmoglea 桥粒芯[桥粒的中央核心层]desmoglein 桥粒芯（糖）蛋白[一类跨膜蛋白，见于桥粒芯]desmolase 碳链酶desmoplakin 桥粒斑蛋白desmosine 锁链素[由四个赖氨酸侧链形成的交联体，见于弹性蛋白]desmosome plaque 桥粒斑desmosome 桥粒desmotubule 连丝微管desmoyokin 桥粒联结蛋白[见于桥粒斑]desolvation 去溶剂化desorption ionization 解析电离desorption 解析（作用），脱附（作用）destabilization 去稳定作用destabilizing agent 去稳定剂destaining 脱色destomycin 越霉素desulfurization 脱硫（作用）desynchronization 去同步化detachment 解析（作用），脱附（作用detection limit 检出限detection 检测，检出；观察detector 检测器detention time 滞留时间detention 滞留detergent 去污剂，除垢剂，洗涤剂deterioration （生物）致劣，恶化determinant 决定簇；决定子[用于胚胎学]；决定因素determination limit 测定限determination 测定detoxication 解毒detoxification 解毒detritus 碎屑；腐屑detritylation 脱（除）三苯甲基deuteromycetes 半知菌纲deutertion 氘化deutoplasm 滋养层，卵黄质developing solvent 展开剂developing tank 展开槽development folder 显影夹developmental biology 发育生物学developmental genetics 发育遗传学developmental malformation 发育畸形developmental phase 发育期developmental regulation 发育调节，发育调控developmental rhythm 发育节律developmental stage 发育阶段developmentally regulated expression 由发育（所）调节的表达devernalization 脱春化dexamethasone 地塞米松，9-alpha-氟-16-甲基脱氢皮质醇dextran bead 葡聚糖珠dextran sulfate 葡聚糖硫酸酯dextran 葡聚糖，右旋糖酐dextranase 葡聚糖酶dextrin 糊精dextroisomer 右旋异构体dextrorotary 右旋的dextrorotatory 右旋的dextrose 葡聚糖diabetes mellitus 糖尿病diacetoxyscirpenol 蛇形菌素diacylglycerol 二酰甘油diad 二分体，二联体diadric 雄异配（性）的diafiltration 渗滤[溶液通过滤器进行连续循环] diagnostic enzyme 诊断酶diagnostic kit 诊断试剂盒diagnostic procedure 诊断程序，诊断手续diagnostic reagent 诊断试剂diagnostics 诊断学diagonal chromatography 对角线层析diagonal electrophoresis 对角线电泳diagonal 对角线；对角线的diagynic 雌异配（性）的diakinesis 终变期dialkylglycine decarboxylase 二烷基甘氨酸脱羧酶dialkylglycine 二烷基甘氨酸diallel cross 双列杂交[用于进化遗传学] dialysate 透析物，透析液dialysis apparatus 透析仪，透析装置dialysis bag 透析袋dialysis cultivation 透析培养dialysis fermentation 透析发酵dialysis membrane 透析膜dialysis tube 透析袋dialysis tubing 透析袋，透析管dialysis 透析dialyzable 可透析的dialyzate 透析液dialyzator 透析仪diamagnetic compound 抗磁化合物diamagnetism 抗磁性diaminedichloroplatinum 二胺二氯铂diaminobenzidine 二氨基联苯胺diaminopimelate 二氨基庚二酸，二氨基庚二酸盐、酯、根diaminopimelic acid 二氨基庚二酸dianisidine 邻联茴香胺dianthovirus 香石竹病毒组[一组植物病毒，模式成员是香石竹环斑病毒]diapause hormone 滞育激素diapedesis 血细胞渗出[血细胞从血管内渗出]diaphragm 隔膜diarrhea 腹泻diastase 淀粉酶制剂diastereomer 非对映（异构）体diastereotopic 非对映异位的[在分子整体中，碳原子上互为非对映关系的原子、基团或面]diastole 心舒期diatomaceous earth 硅藻土diatrizoate 3,5-双（乙酰氨基）-2,4,6-三碘苯甲酸（盐）diauxie growth curve 双峰生长曲线diazo compound 重氮化合物diazo 重氮基diazoacridine 重氮吖啶diazobenzyloxymethyl paper 重氮苄氧甲基纸，DBM纸diazonorleucine 重氮基正亮氨酸diazophenylthio paper 重氮苯硫醚纸，DPT纸diazotization 重氮化diazouridine 重氮尿苷dibucaine 狄步卡因dicarboxyl cellulose 二羧基纤维素dicarboxylic acid 二羧酸dicarboxylic amino acid 二羧基氨基酸dicentric chromosome 双着丝染色体dichlorodimethylsilane 二氯二甲硅烷dichlorofluorescein 二氯荧光黄dichloromethane 二氯甲烷dichlorovos 敌敌畏dichogamy 雌雄（蕊）异熟dichroism 二色性Dick test 狄克试验[链球菌红斑毒素的皮肤试验] dicotyledons 双子叶植物dicoumarin 双羟香豆素，败坏翘摇素dictyosome （分散）高尔基体dictyostelium 盘基网柄菌属，网柄菌属dicyclohexylcarbodiimide 二环己基碳二亚胺[常用缩合剂] didanosine [商]2',3’-双脱氧肌苷didehydrothymidine 双脱氢胸苷dideoxy sequencing method 双脱氧测序法dideoxyadenosine triphosphate 双脱氧腺苷三磷酸dideoxycytidine triphosphate 双脱氧胞苷三磷酸dideoxycytidine 双脱氧胞苷dideoxyguanosine triphosphate 双脱氧鸟苷三磷酸dideoxyguanosine 双脱氧鸟苷dideoxyinosine 双脱氧肌苷dideoxyribonucleoside triphosphate 双脱氧核苷三磷酸dideoxyribonucleoside 双脱氧核苷dideoxythymidine triphosphate 双脱氧胸苷三磷酸dielectric constant 介电常数dielectric effect 介电效应dielectrometric titration 介电（常数）滴定（法）dielectrometry 介电（常数）滴定（法）dielectrophoresis 介电（电）泳diene 双烯diethyl pyrocarbonate 焦碳酸二乙酯diethyl sulfate 硫酸二乙酯diethylstilbestrol 乙酚，二乙基己烯雌酚difference spectrum 差光谱differential （示）差的，鉴别的；微分；微分的differential analysis 示差分析differential centrifugation 差速离心differential detection 示差检测，鉴别检测differential expression 差异表达differential flotation centrifugation 差速浮式离心differential hybridization 示差杂交（法）differential medium 鉴别培养基differential operation 示差操作differential permeability 差别透性，选择透性differential precipitation 示差沉淀differential refractive index detector 示差折光率检测器differential scattering 差散射differential screening 示差筛选differential sedimentation 差速沉降differential sepctrophotometry 示差分光光度法differential species 区别种differential spectrum （示）差光谱differential staining technique 鉴别染色技术[有时特指染色体显带技术] differential staining 鉴别染色（法）differential type detector 微分型检测仪differentiating solvent 鉴别剂，区分溶剂differentiation antigen 分化抗原differentiation center 分化中心differentiation phase 分化时differentiation 分化diffraction grating 衍射光栅diffraction symmetry 衍射对称性diffraction 衍射diffuser 扩散器；洗料器；[发酵罐]进气装置diffusion chamber 扩散盒，扩散小室diffusion coefficient 扩散系数diffusion controlled reaction 扩散控制（的）反应diffusion controlled termination 扩散控制的终止diffusion 扩散diffusional limitation 扩散限制diffusional resistance 扩散阻力diformazan 二甲digalactosyl diglyceride 双半乳糖甘油二酯digester 消化器，消化罐digestion 消化，（酶切）消化digestive enzyme 消化酶digital control 数字控制digital imaging microscope 数字成像显微镜digital imaging microscopy 数字成像显微术digitalis cardiac glycoside 毛地黄（类）强心苷digitalizer 数字化仪[用于计算机科学]digitonin 毛地黄皂苷digitoxigenin 毛地黄毒苷配基；beta-(丁烯酸内酯)-14-羟甾醇digitoxin 毛地黄毒苷diglyceride 甘油二酯digoxigenin 洋地黄毒苷，地高辛配基digoxin 异羟基洋地黄毒苷原，地高辛dihaploid 双单倍体dihedral angle 二面角，双面角dihydrobiopterin 二氢生物蝶呤dihydrochalcone 双氢查耳酮，二氢查耳酮dihydrofolate reductase 二氢叶酸还原酶dihydrofolate 二氢叶酸dihydrolipoamide dehydrogenase 二氢硫辛酰胺脱氢酶dihydrolipoamide 二氢硫辛酰胺dihydrolipoic acid 二氢硫辛酸dihydroorotase 二氢乳清酸酶dihydroorotate 二氢乳清酸dihydropteridine reductase 二氢蝶啶还原酶dihydropteridine 二氢蝶啶dihydropyridine 二氢吡啶dihydrotestosterone 双氢睾酮dihydrouracil arm 二氢尿嘧啶臂dihydrouracil loop 二氢尿嘧啶环dihydrouracil 二氢尿嘧啶dihydrouridine 二氢尿苷dihydroxyacetone phosphate 二氢丙酮磷酸dihydroxycholecalciferol 二羟胆钙化（固）醇dihydroxyphenylanaline 二羟苯丙氨酸，多巴dihydroxyphenylethylamine 二羟苯基乙胺，羟酪胺，多巴胺diisopropylfluorophosphate 二异丙基氟磷酸dikaryon 双核体diltiazem 硫氮酮diluent 稀释剂，稀释液dilution cloning 稀释克隆法[如用于获得细胞克隆株] dimensional electrophoresis 双向电泳dimer 二聚体dimerization cofactor 二聚化辅因子dimerization 二聚化dimethoxytrityl 二甲氧三苯甲基[在DNA合成中用作羟基保护剂] dimethyl sulfate 硫酸二甲酯dimethyl sulfoxide 二甲基亚砜dimethylallylpyrophosphate 二甲（基）烯丙基焦磷酸dimethylaminoazobenzene 二甲基氨基偶氮苯dimethylformamide 二甲基甲酰胺dimorphism 二态二氢，双态现象dinitro benzene 二硝基苯dinitrochlorobenzene 二硝基氯苯dinitrofluorobenzene 二硝基氟苯dinitrogen 双氮，分子氮dinitrogenase reductase 固氮酶还原酶dinitrogenase 固氮酶dinitrophenol 二硝基苯酚dinitrophenyl 二硝基苯基dinoflagellate 甲藻dinoxanthine 甲藻黄素dinucleotide frequency 二核苷酸频率dinucletide 二核苷酸diodeelectrode 二极管电极dioecism 雌雄异体，雌雄异株diosgenin 薯蓣皂苷配基，薯蓣皂苷元dioxide 二氧化物dioxygen 双氧dioxygenase 双加氧酶dipeptidase 二肽酶dipeptide 二肽diphenylamine blue 二苯胺蓝diphenyloxazole 二苯基唑diphosphate 二磷酸diphosphatidylglycerol 双磷脂酰甘油diphosphoglycerate shunt 二磷酸甘油酸支路diphosphoglycerate 二磷酸甘油酸diphosphoinositide 二磷酸肌醇磷脂，磷脂酰肌醇磷酸diphthamide 白喉酰胺diphthera toxin 白喉毒素dipicolinic acid 2,6-吡啶二羧酸diplobacillus 双杆菌diploblastic 双胚层的diplococcus pneumoniae 肺炎双球菌diplococcus 双球菌diploid cell line 二倍体细胞系diploid 二倍体diploidization 二倍化diploidy 二倍性diplonema 双线期diplotene stage 双线期dipolar aprotic solvent 偶极非质子溶剂dipolar protophilic solvent 偶极亲质子溶剂dipolar protophobic solvent 极疏质子溶剂dipole molecule 偶极分子dipole moment 偶极矩dipole 偶极dipyrromethane 联吡咯甲烷direct cross 正交direct duplication 同向重复direct insertion 同向插入direct repeat 同向重复（序列）direct selection 正选择[使用只有突变体或重组体能生长的条件进行选择] directed cloning 定向克隆directed mutagenesis 定向诱变directed perturbation 定向微扰directed sequencing 定向测序direction selectivity 方向选择性directional cloning 定向克隆disaccharide 二糖disassembly 解装配，分解disc electrophoresis 圆盘电泳disc gel electrophoresis 圆盘凝胶电泳disc membrane 圆盘膜discharge 放电；卸下discoidal cleavage 盘状卵裂discontinuous epitope 非连续表位discontinuous gradient 不连续梯度discontinuous replication 不连续复制discontinuous variation 不连续变异discontinuous zone electrophoresis 不连续区带电泳discrete 分立的，不连续的discriminant analysis 判别分析discrimination 辨别，判别disease association 疾病相关disease resistance 抗病性dish 平皿disinfectant 消毒剂disinfection 消毒disinfestation 灭虫disinhibition 去抑制disintegration 蜕变，衰变；去整合，解整合；分解，破碎disintegrator 粉碎器，粉碎机disjunction 分离disk centrifuge 圆盘（式）离心机[带有成叠的有孔圆盘，常用于溶液的澄清化处理] dislocation 脱位，转位，位错dismutase 岐化酶disome 二体，双体disordered state 无序状态disordered structure 无序结构dispase 分散酶，中性蛋白酶[用于分散组织培养中的动物细胞]dispenser 分液器dispermy 双精人卵dispersant 分散剂disperse medium 分散介质disperse phase 分散相disperse system 分散系统dispersion force 分散力；色散力dispersion spectrum 色散谱dispersion 分散；色散displaced loop 替代环displacement analysis 顶替（分析）法displacement chromatography 顶替层析displacement electrophoresis 顶替电泳displacement loop 替代环，D环[形如英文字母D] displacement reaction 置换反应displacement 顶替，替代，置换disposable glove 一次性手套disposable microcentrifuge tube 一次性（使用的）微量离心管disposable tip 一次性（使用的）吸头disproportionation 岐化（反应）disrotatory 对旋disruption 破裂，破坏dissecting microscope 解剖显微镜dissection 解剖，剖分disseminated intravascular coagulation 弥漫性血管内凝血dissimilation 异化（作用）dissociation constant 解离常数dissociation 解离，离解dissolvability 溶（解）度，（可）溶（解）性dissolvant 溶剂distamycin 偏端霉素distance receptor 距离感受器distant hybirdization 远缘杂交distant hybrid 远缘杂种distorted peak 畸峰diterpene 双萜，二萜dithioerythritol 二硫赤藓糖醇dithiothreitol 二硫苏糖醇divergence 分散[用于神经系统]；趋异divergent 趋异进化diverse ion effect 异离子效应diversity gene D基因[为D区编码的基因]diversity region 多变区，D区[免疫球蛋白等分子重链的一个高变区] divinylbenzene 二乙烯苯dizygotic twins 二卵双生，异卵双生DNA adduct DNA加合物DNA amplification in vitro DNA体外扩增DNA amplification polymorphism DNA扩增多态性DNA bending DNA转折，DNA弯曲DNA blotting DNA印迹（法）DNA catenation DNA连环DNA circle DNA环[指环状DNA]DNA cleavage DNA裂解，DNA切割DNA cloning DNA克隆（化）DNA jumping technique DNA跳查技术DNA ladder DNA梯[如大小不同的标准参照物的电泳谱]DNA nicking DNA切口形成DNA pitch DNA螺距DNA sizing DNA大小筛分DNA sizing gene DNA大小决定基因[如见于噬菌体，可决定所包装的DNA量]DNA typing DNA分型DNAase DNA酶DNAase I footprinting DNA酶足迹法docking protein 船坞蛋白，停靠蛋白[内质网上与信号识别颗粒相互作用从而使蛋白质继续翻译的蛋白]docking 停靠dodecahedron 十二面体dodecane 十二烷dodecapeptide motif 十二肽基序dodecyl 十二烷基dolichol 多萜醇，长醇domain assmbly 结构域装配domain deletion 结构域删除domain substitute 结构域置换domain 域，区域，结构域，功能域dominance variance 显性方差dominance 显性；优势（度）dominant acting gene 显性开放基因dominant allele 显性等位基因dominant gene 显性基因dominant hemisphere 优势半球dominant interference 显性干涉dominant lethal 显性致死dominant mutation 显性突变dominant negative mutant 显性失活突变体dominant negative 显性阴性的，显性失活的dominant oncogenic 显性致癌的dominant 显性的，优势的Donnan dialysis 唐南透析Donnan equilibrium 唐南平衡Donnan potential 唐南膜电势donor splicing site 剪接供体donor 供体，给体dopamine 多巴胺dosage compensation 计量补偿（效应）dot blot 斑点印迹，斑点印迹膜dot blotting 点渍法，斑点印迹（法）dot hybridization 斑点杂交dotting 打点，打点杂交double antibody method 双抗体法[免疫测定方法之一种]double balloon catheter 双气囊导管double bar 重棒眼，双棒眼，超棒眼[黑腹果蝇唾液腺染色体的X染色体上16A区段重复三次而出现的特殊表型]double beam mass spectrometer 双束质谱仪double beam spectrophotometer 双光束分光光度计double blind trial 双盲试验double bond migration 双键移位double coupling method 双偶联法，双偶合法double decomposition reaction 复分解double exchange 双交换double fertilization 双受精double focusing mass spectrometer 双聚焦质谱仪double focusing 双聚焦double helix 双螺旋double immunodiffusion 双向免疫扩散，免疫双扩散double innervation 双重神经支配double labeling 双重标记double minute chromosome 双微染色体[所携带基因得到扩增的成对额外小染色体] double recessive 双隐性double resonance 双共振doublet 双联体；双峰doubling time 倍增时间[培养物的生物质翻一番所需的时间]Dower resin Dower树脂[陶氏化学公司离子交换层析介质商品名]down promoter mutation 启动子减效突变downflow fixed bed 下流固定床doxorubicin 阿霉素drift （遗传）漂变drilling mud 钻探泥浆drinking center 饮水中枢drop method 点滴法droplet countercurrent chromatography 液滴反流层析，液滴逆流层析dropping mercury electrode 滴汞电极Drosophila 果蝇属drug susceptibility 药物敏感性drug targeting 药物寻靶，药物导向Duchenne muscular dystrophy Duchenne型肌营养不良，假肥大型肌营养不良duocrinin 促十二指肠液素duplex 双链体；双螺旋；二显性组合duplicon 重复子duramycin 耐久霉素dwarf colony 侏儒型菌落dwarf plant 矮化植物[由遗传因素决定不能长高]；矮生植物[由认为措施或特殊环境决定不能长高]dyad symmetry 二重对称dyad 二分体，二联体dye exclusion test 染料排斥试验[用于检查细胞生活力]dynactin 动力蛋白激活蛋白dynamin 发动蛋白dynein arm 动力蛋白臂dynein 动力蛋白dynorphin 强啡肽dysbacteria 菌群失调dysbacteriosis 菌群失调dysentery bacillus 痢疾杆菌dysfunction 功能异常，机能障碍dysregulation 调节异常dystroglycan （肌）营养不良（蛋白）聚糖[与肌）营养不良蛋白相关的蛋白聚糖] dystrophin (肌）营养不良蛋白。
高效液相色谱法测定吲哚美辛控释胶囊的血药浓度和生物利用度
高效液相色谱法测定吲哚美辛控释胶囊的血药浓度和生物利用
度
张丹;曾经泽;边巴仓决
【期刊名称】《色谱》
【年(卷),期】1997(15)6
【摘要】采用ＯＤＳ柱，甲醇－知磷酸溶液为流动相，２６０ｎｍ为检测波长，建立了测定血浆中吲哚美辛浓度的高效液相色谱法，并测定了吲哚美辛控释胶囊炎痛康的血药浓度。
结果表明，血浆中哚美辛浓度在０．１２５－５．０ｍｇ／Ｌ范围内线性关系良好，检测限６２．５μｇ／Ｌ，
【总页数】1页(P515)
【作者】张丹;曾经泽;边巴仓决
【作者单位】华西医科大学药学院;华西医科大学药学院
【正文语种】中文
【中图分类】R971.1
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2.高效液相色谱法测定氨糖美辛肠溶片中吲哚美辛含量 [J], 逯小萌
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蒋学华
5.吲哚美辛控释制剂的相对生物利用度 [J], 吴苏澄;姜云平;钟诗龙;曾仁杰;蒋学华因版权原因，仅展示原文概要，查看原文内容请购买。
丹参素钠大孔树脂吸附分离过程的近红外在线监测研究
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