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

• Published in: Journal of pharmaceutical and biomedical analysis Vol. 53; no. 1; pp. 90 - 97
• Main Authors: Almeida, B.E., Oliveira, J.E., Carvalho, C.M., Dalmora, S.L., Bartolini, P., Ribela, M.T.C.P.
• Format: Journal Article
• Language: English
• Published: Amsterdam Elsevier B.V 21-09-2010; Elsevier
• Subjects: Analysis; Analytical, structural and metabolic biochemistry; Biological and medical sciences; Chorionic Gonadotropin - analysis; Chorionic Gonadotropin, beta Subunit, Human - analysis; Chromatography, High Pressure Liquid - methods; Chromatography, Reverse-Phase - methods; Fundamental and applied biological sciences. Psychology; General pharmacology; Glycoprotein Hormones, alpha Subunit - analysis; hCG; hLH; hMG; Humans; Luteinizing Hormone - analysis; Medical sciences; Pharmaceutical Preparations - analysis; Pharmacology. Drug treatments; Recombinant Proteins - analysis; RP-HPLC; Subunits; RP-HPLC; hMG; hLH; hCG; Subunits; Human; HPLC chromatography; Subunit; Placental hormone; Protein hormone; Human chorionic gonadotrophin; Reversed phase chromatography; Adenohypophyseal hormone; Analysis; Preparation; Luteinizing hormone; Chorionic gonadotrophin
• ISSN: 0731-7085; 1873-264X
• DOI: 10.1016/j.jpba.2010.03.013

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 α- and β-subunits migrated with significantly different retention times (
t
R) in the following order of increasing hydrophobicity: α-hCG
<
α-hLH
<
hCG
<
hLH
<
β-hCG
<
β-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.