Isotope-dilution method for the determination of 1-vinyl-2-pyrrolidone-mercapturic acid as a potential human biomarker for 1-vinyl-2-pyrrolidone via online SPE ESI-LC/MS/MS in negative ionization mode.
We established and validated a specific and sensitive analytical method for the determination of 1-vinyl-2-pyrrolidone (VP) as 1-vinyl-2-pyrrolidone-mercapturic acid (VPMA) in urine using an electrospray liquid chromatography tandem mass spectrometry (ESI-LC/MS/MS) column switching method. An online solid phase extraction (SPE) for sample cleanup was performed by column switching to a restricted access material and back-flushing to the analytical column. A Phenomenex Luna C8 column was used for sample separation (150mm; ID 4,6mm; 3μm). D4-VPMA served as an isotope labeled internal standard and was detected in negative multiple-reaction monitoring (MRM) mode. The Limit of quantification (LOQ) for VPMA was 1.5μg/L, the intra-day precision of three concentrations (2μg/L, 75μg/L and 400μg/L) of spiked urine samples ranged from 2.7 to 7.3%, the inter-day precision from 3.4 to 14.4%. The accuracy ranged from 6.2 to 9.0%, for the intra-day experiments and from 0.3 to 6.9% for the inter-day experiments. The method was applied to urines of Sprague-Dawley rats exposed to VP as a proof of principle of VPMA as a potential biomarker.
Institute for Occupational and Social Medicine, Medical Faculty, RWTH Aachen University, Pauwelsstr. 30, D-52074 Aachen, Germany. Electronic address: jbertram@ukaachen.de.
J
J
Bertram
Institute for Occupational and Social Medicine, Medical Faculty, RWTH Aachen University, Pauwelsstr. 30, D-52074 Aachen, Germany.
T
T
Schettgen
Institute for Occupational and Social Medicine, Medical Faculty, RWTH Aachen University, Pauwelsstr. 30, D-52074 Aachen, Germany.
T
T
Kraus
D000111:Acetylcysteine / Q000031:analogs & derivatives* / Q000737:chemistry / Q000652:urine; D000818:Animals; D015415:Biomarkers / Q000652:urine; D002138:Calibration; D002853:Chromatography, Liquid / Q000379:methods; D004355:Drug Stability; D006801:Humans; D007553:Isotope Labeling / Q000379:methods*; D009862:Online Systems; D011760:Pyrrolidinones / Q000737:chemistry / Q000652:urine*; D017207:Rats, Sprague-Dawley; D015203:Reproducibility of Results; D052616:Solid Phase Extraction / Q000379:methods*; D012996:Solutions; D021241:Spectrometry, Mass, Electrospray Ionization / Q000379:methods*; D053719:Tandem Mass Spectrometry / Q000379:methods*
C000614238:1-vinyl-2-pyrrolidonemercapturic acid; D015415:Biomarkers; D011760:Pyrrolidinones; D012996:Solutions; C042670:N-vinyl-2-pyrrolidinone; C028537:2-pyrrolidone; D000111:Acetylcysteine
1-Vinyl-2-pyrroldione; Biomarker; LC/MS/MS; Mercapturic acid; Urine