人促卵泡激素效价测定的初步研究——反相高效液相色谱法与体内生物测定法的比较。

A pilot study on potency determination of human follicle-stimulating hormone: a comparison between reversed-phase high-performance liquid chromatography method and the in vivo bioassay.

机构信息

Biotechnology Department, IPEN-CNEN, Av. Prof. Lineu Prestes 2242, Cidade Universitária, São Paulo, Brazil.

出版信息

J Pharm Biomed Anal. 2011 Mar 25;54(4):681-6. doi: 10.1016/j.jpba.2010.10.018. Epub 2010 Oct 30.

DOI:10.1016/j.jpba.2010.10.018

Abstract

Reversed-phase high-performance liquid chromatography (RP-HPLC) was compared with the classical Steelman-Pohley bioassay (BA), based on animal use, for the determination of human follicle-stimulating hormone (hFSH) biological activity. A linear relationship (BA(IU)=0.9925 RP-HPLC(IU)-1.3165) with a highly significant correlation (r=0.9371; p<0.0001; n=24) was found for these two methods for six hFSH preparations of different origins. The mean difference between the bioactivity predicted from RP-HPLC data via this equation and the mean of the bioactivities obtained with the two methods for six other hFSH preparations was -1.4%, with a 95% confidence interval of -9.3 to +6.6%. The precision of these parameters was 1.63% and 2.82%, respectively. These results demonstrate that RP-HPLC is a viable physical-chemical alternative to the use of an in vivo bioassay for hFSH potency determination, applicable also to hFSH Standards containing large amounts of human serum albumin.

摘要

反相高效液相色谱法（RP-HPLC）与经典的 Steelman-Pohley 生物测定法（BA）进行了比较，后者基于动物使用，用于测定人促卵泡激素（hFSH）的生物活性。对于六种不同来源的 hFSH 制剂，这两种方法之间存在线性关系（BA(IU)=0.9925 RP-HPLC(IU)-1.3165），相关性非常显著（r=0.9371；p<0.0001；n=24）。通过该方程从 RP-HPLC 数据预测的生物活性与使用这两种方法获得的六种其他 hFSH 制剂的生物活性的平均值之间的平均差异为-1.4%，95%置信区间为-9.3 至+6.6%。这些参数的精度分别为 1.63%和 2.82%。这些结果表明，RP-HPLC 是一种可行的物理化学替代方法，可替代使用体内生物测定法来测定 hFSH 效力，也适用于含有大量人血清白蛋白的 hFSH 标准品。