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.

Biochemical and hormonal parameters analysis of metoclopramide-induced hyperprolactinemic female wistar rats administered leaf extract of Kigelia africana

“…Follicle stimulating hormone (FSH) is essential for gonadal development and maturation at puberty, as well as gamete production. It stimulates the growth and maturation of ovarian follicles by acting directly on the receptors located on the granulosa cells [2].The significant reduction of FSH level of metoclopramide-induced hyperprolactinemia control (HPT-CON) probably, may be due to gonadal dysfunction which delays maturation of ovarian follicles in the pre-ovulatory phase. Assessments of Fig.…”

“…Some factors such as genetic, anatomic, hormonal and immunological problems are implicated. Secondary sources like sexually transmitted infections, surgery and obesity cannot be ruled out [2]. Prolactin is a peptide hormone secreted by the anterior pituitary gland which plays a pivotal role in a variety of reproductive functions.…”

Physical-chemical and biological characterization of different preparations of equine chorionic gonadotropin

“…The identity and purity of the commercial and reference preparations of eCG were investigated by RP-HPLC using a methodology derived from those previously established in our laboratory for the analysis of human LH, FSH and hCG, which have confirmed the system precision [342129]. A Shimadzu Model SCL-10AHPLC apparatus with a SPD-10AV UV detector and a Vydac 214TP C 4 54 column (25 cm × 4.6 mm; pore diameter 300 Å and particle diameter of 5 µm; Grace, USA) coupled to a Vydac 214FSK 54 guard pre-column (1 cm × 4.6 mm; Grace) was used.…”

Absolute Quantification of Follicle Stimulating Hormone (FSH) Using Its Signature Peptides and Enzymatic Digestion in Human Serum by UPLC/LC–MS/MS

“…The quantification of FSH holds clinical relevance, especially in controlled ovarian stimulation regimens for assisted reproductive technologies and hypogonadotrophic hypogonadism. Traditionally, useful information concerning purity, identity, potency, and stability assessment of FSH and pituitary hormones, in general, has been provided using high-performance liquid chromatography (HPLC) and commercially by Ligand binding assays or immunoassays [4,5]. Ligand binding assays though with a sensitivity of as low as in mIU mL −1 level, require the development of suitable capture and detection reagents, which takes time and resources, making them less affordable in drug discovery and early development.…”