胰岛素生物检定的快速测定方法的建立

目的:建立胰岛素生物测定的快速检测方法。方法:血糖试纸上的葡萄糖脱氢酶将血液中的葡萄糖转化为葡萄糖酸内脂。这种作用会产生直流电,血糖仪以此判读血糖值。使用血糖仪快速测定小鼠血糖。结果:血糖仪能够快速测定小鼠血糖值,与全自动生化分析仪测定结果高度相关,效价偏差率<5%,结果无显著性差异(P>0.05)。结论:经方法学验证,本法可用于胰岛素生物活性的快速测定。     

Larvicidal activity of Persea americana Mill. against Aedes aegypti

ObjectiveTo evaluate the toxicity of the ethanol and hexane extracts of the different parts of Persea americana Mill. (P. americana) toward third and fourth instars larvae of Aedes aegypti (Ae. aegypti) and to characterize the ethanol extract by qualitative phytochemical analysis.MethodsThe seeds, peels and pulp of P. americana were processed for crude extraction using 95% ethanol and n-hexane. Crude extracts were bio-assayed for larvicidal activity against Ae. aegypti following the World Health Organization standard bioassay method. The mortality was observed at 24 h and 48 h after treatment and data were subjected to probit analysis to determine lethal concentrations (LC₅₀ and LC₉₀). The ethanol extract was characterized by phytochemical analysis.ResultsBoth the hexane and ethanol extracts from the different parts of P. americana exhibited evidence of larvicidal toxicity. The hexane extract from the seeds exhibited the highest toxicity with LC₅₀ and LC₉₀ values of 9.82 mg/L and 22.19 mg/L, respectively, while the ethanol seed extract exhibited LC₅₀ of 16.48 mg/L and LC₉₀ 45.77 mg/L, respectively. This was closely followed by the ethanol extract of the peels with an LC₅₀ of 10.35 mg/L and LC₉₀ of 26.29 mg/L. The pulp extracted with ethanol also yielded great larvicidal toxicity with LC₅₀ of 21.32 mg/L and LC₉₀ of 59.45 mg/L. Results of the phytochemical analysis of the ethanol seed extract indicated presence of alkaloids, tannins, saponins, unsaturated steroids and triterpenoids, flavonoids (leucoanthocyanins), fats and oils.ConclusionsBoth the hexane and ethanol extracts of P. americana showed promising potential as an alternative source of a more sustainable, non-toxic and environmentally friendly solution for the control of dengue vector, Ae. aegypti.     

川芎及其中成药抗凝血作用测定方法的研究

目的以延长活化部分凝血活酶时间(APTT)为指标,建立定量测定川芎抗凝血作用的方法,评价川芎及其中成药的质量。方法川芎先后用乙醇和水定量提取,以总提取物为供试品,体外测定延长APTT值。家兔心脏取血,制备血浆,加入APTT试剂后,测定凝结时间。以APTT延长率为抗凝血活性的评价指标,用阿魏酸钠标定川芎的延长APTT活性。根据量反应平行线法(2.2)计算川芎的抗凝血活性。并测定了9份川芎药材、饮片及中成药的抗凝血活性。结果阿魏酸钠和川芎总提取物均具有显著的抗凝血活性,且可靠性检验结果成立。阿魏酸钠的给药质量浓度(1~5 mg/m L)与其APTT延长率呈良好的线性关系(r=0.995 5)。供试品重复测定抗凝血活性的RSD为9.34%(n=6),可信限率为15.98%(n=6)。不同川芎样品的抗凝血活性不同,5份川芎药材的抗凝血效价分别为5.431~7.620 U/g,川芎饮片及川芎酒炙饮片分别为5.910、3.017 U/g,通脉颗粒和血府逐瘀丸分别为14.516、29.035 U/g。结论建立的方法能准确测定川芎药材、饮片及其中成药的抗凝血活性并评价其质量。     

胰岛素生物测定方法的比较

通过查阅中、美、英、日四国药典中胰岛素生物测定方法的相关内容,对其历年来的生物活性测定方法进行分析与对比,结果各国药典中胰岛素的生物测定方法还存在差异,胰岛素生物测定方法的发展方向与改进空间应参考国外药典统一为HPLC法。     

阿奇霉素胶囊的人体相对生物利用度研究

目的研究国产阿奇霉素胶囊的相对生物利用度。方法采用双周双制剂交叉试验设计，让20名健康男性志愿者随机单剂量口服国产四川维奥阿奇霉素胶囊和四川明欣阿奇霉素胶囊，剂量均为500mg，剂间间隔为2周。用微生物杯碟法测定血清中阿奇霉素浓度，并计算相对生物利用度。结果受试制剂和参比制剂的药代动力学参数无统计学差异，消除半衰期（t1/2B）分别为（53．29±19．33）h和（51．21±17．04）h，峰浓度（Cmax）分别为（0．360±0．118）μg／mL和（0．367±0．132）μg／mL，达峰时间（tmax）分别为（2．30±0．67）h和（2．30±0．86）h，AUC0-τ，分别为（5．42±1．89）μg·h／mL和（5．84±2．29）μg·h／mL。结论受试制剂阿奇霉素胶囊的相对生物利用度为（92．81±11．79）％，与参比制剂具有生物等效性。     

多肽类药物体内分析方法

多肽类药物是新药研究的新领域,也是当今新药研究最活跃和发展最迅速的领域之一。目前,建立测定生物基质中多肽类药物的方法学是世界范围存在的难题,面临着诸多挑战。酶联免疫法(ELISA)及放射同位素标记是定量多肽类药物最常用的方法,应用广泛。但液相色谱-质谱联用技术发展迅速,高专属性、高灵敏度、应用广泛、简便快速等特点势必使其成为多肽类药物体内定量分析的重要手段。本文就目前国际上几种检测方法的优劣、应用现状及发展趋势进行了综述。     

生物检定方法控制中药质量的思考

质量稳定可控是保证中药有效性和安全性的重要前提。现行的中药质控模式和方法仅能控制部分指标性成分的一致性和稳定性，难以全面、有效地控制中药的质量。而生物检定方法直接关切有效性和安全性，特别适用于结构复杂或者理化测定不能反映其临床疗效的药物，在中药质量控制和评价中具有独到的优势。在现行化学控制基础上建立中药生物检定质控方法，比目前主要基于指标性成分定性定量分析的质控体系具有更大的实际意义和优势，亦将成为中药质量控制发展的新趋势。     

高效液相色谱法和生物测定法用于测定血清中环丙氟哌酸浓度的比较

采用ＨＰＬＣ测定血清中环丙氟哌酸的浓度。流动相为１４％甲醇，８％乙腈，７８％磷酸盐缓冲液（其中含０．３％氢氧化四丁基铵，０．０２ｍｏｌ／Ｌ磷酯二氢钾，ｐＨ＝３．０±０．０２）。采用反相色谱柱，２７７ｎｍ紫外检测。二氯甲烷提取，以二氟沙星为内标物，血样中药物检测限是１０ｎｇ／ｍｌ，回收率９８．８％，日内相对标准偏差是３．５％，日间相对标准偏差是１．３８％。通过兼用ＨＰＬＣ和生物检定法测定健康受试者的血清药物浓度，对两种分析方法进行比较，将用两种方法测定的结果（ｎ＝８０）进行线性回归，直线的斜率是０．９１，相关系数是０．９５９（Ｐ＜０．０１）。     

A rapid sperm motility bioassay procedure for quality-control testing of water and culture media

A rapid bioassay procedure is described for quality-control testing water and apparatus used in the preparation of media for gamete and embryo culture. This bioassay is based on the sensitivity of hamster epididymal spermatozoa to contaminants present in water and/or in the culture apparatus. The bioassay is usually performed using a modified Tyrode's solution as the sperm culture medium, although complex media can be used. The sensitivity of this test is greatly enhanced by the absence of protein in the medium. The bioassay has been used to detect impurities in water prepared by a standard cartridge filtration system and to verify that reverse-osmosis pretreatment of water could eliminate the problem. It has also detected toxic contaminants leached from syringe filters during medium sterilization. The bioassay is simple to perform and can be completed in 1 working day. It may be a useful alternative to the conventional mouse embryo tests that are in widespread use in human in vitro fertilization (IVF) laboratories.     

Mineral deficiency and the use of the FETAX bioassay to study environmental teratogens

The Frog Embryo Teratogenesis Assay: Xenopus (FETAX) bioassay has been employed extensively to screen compounds for teratogenic activity. Recent laboratory studies have indicated that low potassium concentrations retard Xenopus laevis development. The effects of varying concentrations of minerals on Xenopus laevis embryo length and development were examined to determine the utility of the FETAX bioassay in the study of environmental teratogens. Water samples collected from 18 wetlands in Minnesota and North Dakota correlated with low mineral levels, causing delayed embryonic development in the FETAX bioassay. When the concentration of sodium or potassium was <5 ppm, Xenopus laevis development was delayed. As a result, it was difficult to assess teratogenic activity after 96 h of incubation. Furthermore, the length of the embryos-an indication of development-paralleled changes in mineral composition. Comparisons between different wetlands based on changes in one specific mineral were not possible due to a synergism between various minerals. If the concentration of sodium and/or potassium was <5 ppm but > or =2 ppm, extension of the FETAX bioassay to 120 h allowed organogenesis to proceed through stage 46, as required for scoring in accordance with ASTM guidelines for the FETAX bioassay. In those cases in which the concentration of sodium and/or potassium were <2 ppm, the embryos could not develop to stage 46 within 120 h and the FETAX bioassay was not suitable for detecting teratogenic activity. Published in 2002 by John Wiley & Sons, Ltd.