Premium
Determination of S ‐phenylmercapturic acid in human urine using an automated sample extraction and fast liquid chromatography‐tandem mass spectrometric method
Author(s) -
Li Yinghe,
Li Austin C.,
Shi Haihong,
Junga Heiko,
Jiang Xiangyu,
Naidong Weng,
Lauterbach John H.
Publication year - 2006
Publication title -
biomedical chromatography
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.4
H-Index - 65
eISSN - 1099-0801
pISSN - 0269-3879
DOI - 10.1002/bmc.653
Subject(s) - chemistry , chromatography , calibration curve , sample preparation , extraction (chemistry) , selected reaction monitoring , urine , electrospray ionization , mass spectrometry , detection limit , analytical chemistry (journal) , tandem mass spectrometry , biochemistry
S ‐phenylmercapturic acid is widely accepted as a specific biomarker for the evaluation of benzene exposure. Here, we describe a fast, specific and sensitive high‐performance liquid achromatography coupled with tandem mass spectrometry (LC‐MS/MS) method that has been developed and validated for the determination of S‐ phenylmercapturic acid in human urine. Isotope‐labeled S‐ phenylmercapturic acid‐ d 5 was used as internal standard to improve the method ruggedness. The fully automated solid‐phase extraction method on a 96‐well Oasis MAX (mix‐mode anion exchange) plate was employed to clean up the urine samples before analysis. The rapid LC‐MS/MS analysis of extracted samples was achieved on a Genesis C18 column with a run time of only 3 min. Negative electrospray ionization with multiple reaction monitoring (ESI‐MRM) mode was used to detect S‐ phenylmercapturic acid ( m/z 238 → 109) and S‐ phenylmercapturic acid ‐ d 5 ( m/z 243 → 114). The method fulfils all the standard requirements of method validation. The calibration curve was linear within the concentration range 0.400–200 ng/mL. The method performed accurately and precisely in validation with <7.5% relative error and <6.5% relative standard deviation of quality control samples. The method efficacy was also verified by the analysis of urine samples from 12 smokers and 12 non‐smokers. With the fully automated sample cleanup procedure and the fast LC‐MS/MS analysis, a sample analysis throughput of 384 samples per day could be achieved. Copyright © 2006 John Wiley & Sons, Ltd.