Assays - 3-HPMA

High-performance liquid chromatographic-tandem mass spectrometric determination of 3-hydroxypropylmercapturic acid in human urine

Author: D.G. Mascher, H.J. Mascher, G. Scherer, E.R. Schmid
Publisher: J. Chromatogr. B, 750 (2001), 163 - 169


A sensitive and specific HPLC-tandem MS (HPLC-MS/MS) method was developed for the determination of 3-Hydroxypropylmercapturic acid in human urine. Samples were extracted using ENV+ cartridges and then injected on to a C8 Superspher Select B column with acetonitrile / 20 mM formic acid (1:19) as mobile phase. N-acetylcysteine was used as internal standard for HPLC-MS-MS. Linearity was obtained in the tested range of 50 - 5000 ng/ml of urine, with 50 ng/ml as the limit of quantification. Precision, as RSD, in the tested range of 50-5000 ng/ml was 1.47 - 6.04 %. Accuracy ranged from 87 to 114 %. 3-Hydroxypropylmercapturic acid was stable in human urine at 37°C for 24 h. The method was able to quantify 3-Hydroxypropylmercapturic acid in urine of non-smokers and smokers.