title: 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.
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.
authors.affiliation: Biotechnology Department, IPEN-CNEN, Av. Prof. Lineu Prestes 2242, Cidade Universitária, São Paulo, Brazil.
authors.forename: B E
authors.initials: BE
authors.lastname: Almeida
authors.forename: J E
authors.initials: JE
authors.lastname: Oliveira
authors.forename: R
authors.initials: R
authors.lastname: Damiani
authors.forename: S L
authors.initials: SL
authors.lastname: Dalmora
authors.forename: P
authors.initials: P
authors.lastname: Bartolini
authors.forename: M T C P
authors.initials: MT
authors.lastname: Ribela
mesh_terms: D000465:Algorithms; D000826:Animal Testing Alternatives; D000818:Animals; D001681:Biological Assay; D002626:Chemistry, Pharmaceutical; D002851:Chromatography, High Pressure Liquid; D005260:Female; D043373:Follicle Stimulating Hormone, Human / Q000032:analysis* / Q000494:pharmacology*; D006801:Humans; D009929:Organ Size / Q000187:drug effects; D010053:Ovary / Q000187:drug effects / Q000254:growth & development; D010865:Pilot Projects; D011786:Quality Control; D051381:Rats; D017207:Rats, Sprague-Dawley; D011994:Recombinant Proteins / Q000032:analysis / Q000494:pharmacology; D015203:Reproducibility of Results; D012709:Serum Albumin; D013678:Technology, Pharmaceutical
chemical_list: D043373:Follicle Stimulating Hormone, Human; D011994:Recombinant Proteins; D012709:Serum Albumin