Analysis of human luteinizing hormone and human chorionic gonadotropin preparations of different origins by reversed-phase high-performance liquid chromatography.
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.
Almeida
B E
BE
Biotechnology Department, IPEN-CNEN, Av. Prof. Lineu Prestes 2242, Cidade Universitária, 05508-900 São Paulo, Brazil.
Oliveira
J E
JE
Carvalho
C M
CM
Dalmora
S L
SL
Bartolini
P
P
Ribela
M T C P
MT
D006063:Chorionic Gonadotropin / Q000032:analysis*; D018997:Chorionic Gonadotropin, beta Subunit, Human / Q000032:analysis; D002851:Chromatography, High Pressure Liquid / Q000379:methods*; D056148:Chromatography, Reverse-Phase / Q000379:methods*; D015292:Glycoprotein Hormones, alpha Subunit / Q000032:analysis; D006801:Humans; D007986:Luteinizing Hormone / Q000032:analysis*; D004364:Pharmaceutical Preparations / Q000032:analysis*; D011994:Recombinant Proteins / Q000032:analysis
D006063:Chorionic Gonadotropin; D018997:Chorionic Gonadotropin, beta Subunit, Human; D015292:Glycoprotein Hormones, alpha Subunit; D004364:Pharmaceutical Preparations; D011994:Recombinant Proteins; D007986:Luteinizing Hormone