title: Determination of the maleic acid in rat urine and serum samples by isotope dilution-liquid chromatography-tandem mass spectrometry with on-line solid phase extraction.
abstract: A rapid and simple on-line solid-phase extraction coupled with isotope dilution-liquid chromatography-tandem mass spectrometry (SPE-ID-LC-MS/MS) method was developed to quantitate maleic acid in serum and urine of SpragueDawley (SD) rats. The aforementioned biological samples were spiked with (13)C2-maleic acid, vigorously vortexed, added with acetonitrile to precipitate proteins, and then injected into the on-line SPE-LC-MS/MS system for quantification. Upon validation, this method demonstrated excellent feasibility and sensitivity: calibration curves for maleic acid in serum and urine display excellent linearity with the coefficient of determination (R(2)) greater than 0.999; the limits of detection and quantitation (LOD and LOQ) for maleic acid were determined at 0.2 and 0.5μg L(-1), respectively. Additionally, intra-day accuracy for maleic acid in serum and urine samples ranged from 94.0% to 100.2% and 101.3% to 104.4%, respectively. Furthermore, inter-day accuracy ranged from 93.6% to 101.0% and from 102.3% to 111.4% in serum and urine samples, respectively. Intra-day precision %RSD of maleic acid in serum and urine samples was 13.8% or less, whereas the inter-day precision was 6.1% or less. The matrix effects were not found to be statistically significant (p=0.9145 and p=0.5378, correspondingly) based on the calculations of recovery functions. The collected serum and urine samples were analyzed using SPE-ID-LC-MS/MS. Our results reveal trace levels of maleic acid in the control rats, demonstrating that this method is capable of analyzing background levels of contaminants in biofluids with excellent sensitivity and specificity at part-per-billion levels concentrations in complex matrices.
authors.affiliation: Institute of Occupational Medicine and Industrial Hygiene, College of Public Health, National Taiwan University, No. 17, ShiuJou Road, Taipei 10055, Taiwan; National Environmental Health Research Center, National Health Research Institutes, No. 35 , Keyan Road, Zhunan, Miaoli County 35053, Taiwan.
authors.forename: Hsin-Chang
authors.initials: HC
authors.lastname: Chen
authors.affiliation: Institute of Occupational Medicine and Industrial Hygiene, College of Public Health, National Taiwan University, No. 17, ShiuJou Road, Taipei 10055, Taiwan.
authors.forename: Charlene
authors.initials: C
authors.lastname: Wu
authors.affiliation: Institute of Occupational Medicine and Industrial Hygiene, College of Public Health, National Taiwan University, No. 17, ShiuJou Road, Taipei 10055, Taiwan; National Environmental Health Research Center, National Health Research Institutes, No. 35 , Keyan Road, Zhunan, Miaoli County 35053, Taiwan. Electronic address: kuenyuhwu@ntu.edu.tw.
authors.forename: Kuen-Yuh
authors.initials: KY
authors.lastname: Wu
mesh_terms: D000818:Animals; D002853:Chromatography, Liquid / Q000379:methods; D008297:Male; D008298:Maleates / Q000097:blood* / Q000493:pharmacokinetics / Q000652:urine*; D017207:Rats, Sprague-Dawley; D015203:Reproducibility of Results; D052616:Solid Phase Extraction; D053719:Tandem Mass Spectrometry
chemical_list: D008298:Maleates; C030272:maleic acid
keywords: Isotope dilution method; Liquid chromatography–tandem mass spectrometry; Maleic acid; On-line solid-phase extraction; Serum; Urine