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Immunoaffinity extraction and tandem mass spectrometric analysis of human chorionic gonadotropin in doping analysis
Lay-Harn Gam1, Sock-Ying Tham, Aishah Latiff
1School of Pharmaceutical Sciences, University Science of Malaysia, 11800 Minden, Penang, Malaysia. layharn@usm.my
Abstract:
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.