Determination of 3-amino-5-mercapto-1,2,4-triazole in serum.
J Chromatogr B 2004 Mar; 801(2):359-362
A high performance liquid chromatography (HPLC) method using fluorescence detection to determine 3-amino-5-mercapto-1,2,4-triazole (AMT) levels in serum has been developed. Sample preparation involved treatment with tributylphosphine (TBP) to reduce disulfides formed during storage, precipitation of proteins with acetonitrile (ACN), and precolumn derivatization using the thiol reactive fluorescent probe monobromobimane (MBB). The conjugate (AMT-MBB) was resolved by gradient elution from a C(18) reversed-phase column. The assay method was linear over a concentration range of 0.78-50 microg/ml and had a limit of detection (LOD) of 0.05 microg/ml AMT (10 microl injection). This method provides a sensitive and specific tool for the determination of AMT in serum and may have potential industrial hygiene application.
Industrial-hygiene; Sampling; Sampling-methods; Proteins; Antioxidants; Occupational-exposure; Environmental-exposure; Analytical-methods; Analytical-chemistry
Analytical Services Branch, M/S L4218, 1095 Willowdale Road, Morgantown, WV 26505-2888, USA
Disease and Injury: Asthma and Chronic Obstructive Pulmonary Disease
Journal of Chromatography B: Analytical Technologies in the Biomedical and Life Sciences