Immunoaffinity extraction and tandem mass spectrometric analysis of human chorionic gonadotropin in doping analysis

• Published in: Journal of chromatography. B, Analytical technologies in the biomedical and life sciences Vol. 792; no. 2; pp. 187 - 196
• Main Authors: Gam, Lay-Harn, Tham, Sock-Ying, Latiff, Aishah
• Format: Journal Article
• Language: English
• Published: Amsterdam Elsevier B.V 25-07-2003; Elsevier Science
• Subjects: Amino Acid Sequence; Aminoacids, peptides. Hormones. Neuropeptides; Analytical, structural and metabolic biochemistry; Biological and medical sciences; Chorionic gonadotropin; Chorionic Gonadotropin - chemistry; Chorionic Gonadotropin - urine; Chromatography, Affinity - methods; Chromatography, High Pressure Liquid - methods; Fundamental and applied biological sciences. Psychology; Glycoproteins; Humans; Molecular Sequence Data; Peptide Mapping; Proteins; Spectrometry, Mass, Electrospray Ionization - methods; Glycoproteins; Chorionic gonadotropin; Human; Urine; Biological fluid; Antibody; Doping; Glycoprotein; HPLC chromatography; Urinary retention; Antigen; Protein hormone; Analysis method; Affinity chromatography; Mass spectrometry MS/MS; Chorionic gonadotrophin; Extraction; Quantitative analysis
• ISSN: 1570-0232; 1873-376X
• DOI: 10.1016/S1570-0232(03)00264-2

A confirmatory and quantitative HPLC–tandem mass spectrometry (MS–MS) method for human chorionic gonadotropin hormone (hCG) at concentrations as low as 5 IU/l following immunoaffinity extraction of the glycoprotein from urine was developed. The extraction method involved retention of urinary hCG in the immunoaffinity column via specific antigen–antibody interaction. A variety of eluents were then used to quantitatively elute hCG from the immunoaffinity column. Qualitative and quantitative analysis of hCG were undertaken using MS–MS by identifying the amino acid sequence of the marker peptide βT5 obtained from hCG by tryptic digestion and the peak areas of three product ions b
6
+, b
9
+ and y
11
+, respectively.