促卵泡激素体内生物活性测定及其各国药典方法比较  

Comparison of the methods for in vivo FSH bioassay in pharmacopoeia of various countries

作　　者：李湛军[1] 李心欣[1] 梁成罡[1] 

机构地区：[1]中国食品药品检定研究院,北京100050

出　　处：《药物分析杂志》2014年第4期727-732,共6页Chinese Journal of Pharmaceutical Analysis

基　　金：十二五重大新药创制--创新药物研究开发技术平台建设(No.2012ZX09304010)

摘　　要：目的：基于长期大量实验数据，对各国药典促卵泡激素（FSH）生物活性测定方法进行梳理、比较。方法：对中国药典2010年版（ChP2010）FSH生物测定法优化，测定国内外多批重组人促卵泡激素（rhFSH）、尿源FSH及复方rhFSH—rhLH产品的体内生物活性，并就动物数、大鼠体重、溶媒中HCG含量、给药剂量、反应值等与EP7．0和USP35-NF30的FSH的生物测定方法进行对比，梳理其异同。结果：ChP2010每组8只动物与EP7．0和USP35-NF30每组5只动物，计算效价结果总平均偏差率为2．3％，总平均可信限率由21．2％升至28．7％，都在生物测定误差允许范围内。ChP2010规定雌大鼠出生19～23d，体重36。50g，体重偏小，实验灵敏度差，体重36～40g的大鼠卵巢增重明显小于40—50g的空白组卵巢增重（P〈0．05）；溶媒中HCG含量也偏低；给药剂量与EP7．0和USP35-NF30相近；卵巢重作为反应值与EP7．0和USP35-NF30器官系数作为反应值不同，两者测定结果间差异无显著性意义。结论：每组5只大鼠即可满足实验要求，减少实验动物用量（3R原则）。大鼠出生21—23d，体重应为40—60g（最大相差10g内），溶媒中HCG含量适当增加至25～30Iu·mL^-1为最佳FSH生物测定实验条件，优化ChP2010的FSH生物测定法。Objective： To sort out and compare the follicle - stimulating hormone （FSH） bioassay methods in different national pharmacopoeia based on a large number of long - term experimental data. Methods： Various batches of prod- ucts of rhFSH, u - FSH, u - FSH - HP and Co rhFSH - rhLH at home and abroad were evaluated by in vivo FSH bioassay method optimized in Chinese pharmacopeia 2010 （ChP 2010）and were compared with the method in Ep 7. 0 and the USP 35 -NF 30 for FSH in aspects of the number of animals, rat body weight, HCG levels in solvent, FSH dose ranges,response values,etc. Results： The average deviation rate was 2. 3% between each group of eight animals in ChP 2010 and each group of five animals in EP 7.0 and the USP 35 -NF 30;the average confidence limit rate was from 21.2% to 28.7% ,all in the allowed error range of bioassay. In ChP,female rats being 19 -23 d old with body weights ranging 36 - 50 g had poorer experimental sensitivity and their weights were lower. The ovarian weight gain in 36 -40 g rats was significantly smaller than that in 40 -50 g rats in blank group（p 〈0. 05）. HCG levels were also lower in solvents ; FSH dose ranges were similar to EP 7.0 and USP 35 - NF 30. In contrast to EP 7. 0 and USP 35 - NF 30 in which the organ coefficient was taken as the response value, the weight of the ovaries was used as the re- sponse value in ChP 2010,but there was no difference between those measurement results significantly. Conclusion： A total of five rats in each group can meet the bioassay requirement of reduction （ the 3R principle）. It is the best method for FSH bioassay that the rat body weight should be 40 -60 g （the difference between the heaviest and the lightest rat is not more than 10 g）on 21 -23 days after birth;HCG levels in solvents should be increased to 25 -30 IU · mL^-1. ChP 2010 for FSH bioassay should be optimized.

关 键 词：重组人促卵泡激素(rhFSH) 促卵泡激素(FSH) 尿促性素 体内生物测定 生物活性 大鼠卵巢增重 中国药典2010年版(ChP 2010) 优化方法 

分 类 号：R917[医药卫生—药物分析学]