Immunoaffinity extraction and tandem mass spectrometric analysis of human chorionic gonadotropin in doping analysis.
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 betaT5 obtained from hCG by tryptic digestion and the peak areas of three product ions b(6)(+), b(9)(+) and y(11)(+), respectively.
Gam
Lay-Harn
LH
School of Pharmaceutical Sciences, University Science of Malaysia, 11800 Minden, Penang, Malaysia. layharn@usm.my
Tham
Sock-Ying
SY
Latiff
Aishah
A
D000595:Amino Acid Sequence; D006063:Chorionic Gonadotropin / Q000737:chemistry / Q000652:urine*; D002846:Chromatography, Affinity / Q000379:methods*; D002851:Chromatography, High Pressure Liquid / Q000379:methods*; D006801:Humans; D008969:Molecular Sequence Data; D010449:Peptide Mapping; D021241:Spectrometry, Mass, Electrospray Ionization / Q000379:methods*
D006063:Chorionic Gonadotropin