促卵泡激素体内生物活性测定及其各国药典方法比较

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（ChP 2010）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EP 7.0和USP 35-NF 30的FSH的生物测定方法进行对比，梳理其异同。 结果： ChP 2010每组8只动物与EP 7.0和USP 35-NF 30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。ChP 2010规定雌大鼠出生19～23 d，体重36～50 g，体重偏小，实验灵敏度差，体重36～40 g的大鼠卵巢增重明显小于40～50 g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EP 7.0和USP 35-NF 30相近；卵巢重作为反应值与EP 7.0和USP 35-NF 30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21～23 d，体重应为40～60 g（最大相差10 g内），溶媒中HCG含量适当增加至25～30 IU·mL^-1为最佳FSH生物测定实验条件，优化ChP 2010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 products 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 response 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.