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

• Published in: Journal of pharmaceutical and biomedical analysis Vol. 54; no. 4; pp. 681 - 686
• Main Authors: Almeida, B.E., Oliveira, J.E., Damiani, R., Dalmora, S.L., Bartolini, P., Ribela, M.T.C.P.
• Format: Journal Article
• Language: English
• Published: Amsterdam Elsevier B.V 25-03-2011; Elsevier
• Subjects: Algorithms; Analysis; Analytical, structural and metabolic biochemistry; Animal Testing Alternatives; Animals; bioactive properties; Bioassay; bioassays; Biological and medical sciences; Biological Assay; Chemistry, Pharmaceutical; Chromatography, High Pressure Liquid; confidence interval; equations; Female; Follicle Stimulating Hormone, Human - analysis; Follicle Stimulating Hormone, Human - pharmacology; follicle-stimulating hormone; Fundamental and applied biological sciences. Psychology; General pharmacology; hFSH; Human follicle-stimulating hormone; human serum albumin; Humans; Medical sciences; Organ Size - drug effects; Ovary - drug effects; Ovary - growth & development; Pharmacology. Drug treatments; Pilot Projects; Potency; Quality Control; Rats; Rats, Sprague-Dawley; Recombinant Proteins - analysis; Recombinant Proteins - pharmacology; Reproducibility of Results; reversed-phase high performance liquid chromatography; RP-HPLC; Serum Albumin; Technology, Pharmaceutical; RP-HPLC; Human follicle-stimulating hormone; Potency; hFSH; Bioassay; Human; HPLC chromatography; Gonadotropin; Determination; Glycoprotein hormone; In vivo; Reversed phase chromatography; Adenohypophyseal hormone; Comparative study; Quantitative analysis; Follicle stimulating hormone
• ISSN: 0731-7085; 1873-264X
• DOI: 10.1016/j.jpba.2010.10.018

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 (BAIU=0.9925 RP-HPLCIU−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.