Analysis of human luteinizing hormone and human chorionic gonadotropin preparations of different origins by reversed-phase high-performance liquid chromatography

作者：Almeida B E; Oliveira J E; Carvalho C M; Dalmora S L; Bartolini P; Ribela M T C P^*

来源：Journal of Pharmaceutical and Biomedical Analysis, 2010, 53(1): 90-97.

DOI：10.1016/j.jpba.2010.03.013

• RP-HPLC
• hLH
• hCG
• hMG
• Subunits

Specific reversed-phase high-performance liquid chromatography conditions are reported for the analysis of recombinant and native human luteinizing hormone (hLH) and human chorionic gonadotropin (hCG) preparations. Heterodimeric hLH, hCG and their alpha- and beta-subunits migrated with significantly different retention times (t(R)) in the following order of increasing hydrophobicity: alpha-hCG < alpha-hLH < hCG < hLH < beta-hCG < beta-hLH. Under these conditions, the main peak of three hCG preparations ran about 4% faster than the average t(R) (38.35 +/- 0.42 min; RSD = 1.1%) of four hLH preparations. Four heterogeneous urinary products were also analyzed, hLH, hFSH and hCG peaks being identified.
Quantitative analysis was validated for the homogeneous preparations and a highly linear dose-response curve (r = 0.99998; p < 0.0001; n = 20) used to assess the accuracy, precision and sensitivity of the analysis. Quantification of the different gonadotropins in the heterogeneous preparations was also carried out, but with limitations in accuracy.

• 出版日期2010-9-21