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Simultaneous Determination of Five Porphyrins in Human Urine and Plasma Using High Performance Liquid Chromatography-Tandem Mass Spectrometry

  • Hur, Yeoun (Department of Analytical Research Service, International Scientific Standards Co. Ltd.) ;
  • Tae, Sookil (Department of Analytical Research Service, International Scientific Standards Co. Ltd.) ;
  • Koh, Yun-Joo (The Korea Institute for children's Social Development and Rudolph Child Research Center) ;
  • Hong, Sung-Hyun (Metabolomics Research Center, Institute for EONE Laboratories) ;
  • Yoon, Young Ho (Metabolomics Research Center, Institute for EONE Laboratories) ;
  • Jang, Haejong (Department of Analytical Research Service, International Scientific Standards Co. Ltd.) ;
  • Kim, Sooji (Department of Analytical Research Service, International Scientific Standards Co. Ltd.) ;
  • Kim, Kyeong Ho (Department of Pharmacy, College of Pharmacy, Kangwon National University) ;
  • Kang, Seung Woo (Department of Analytical Research Service, International Scientific Standards Co. Ltd.) ;
  • Lee, Youngshin (Department of Analytical Research Service, International Scientific Standards Co. Ltd.) ;
  • Han, Sang Beom (Department of Pharmaceutical Analysis, College of Pharmacy, Chung-Ang University)
  • 발행 : 2014.06.30

초록

A specific and sensitive liquid chromatography-electrospray ionization tandem mass spectrometry method (LC-ESI-MS/MS) was developed and validated for the simultaneous quantification of porphyrins (coproporphyrin, pentacarboxylporphyrin, hexacarboxylporphyrin, heptacarboxylporphyrin, and uroporphyrin) in human plasma and urine. Acidified plasma samples and urine samples were prepared by using liquid-liquid extraction using ethyl acetate and protein precipitation with acetonitrile, respectively. The separation was achieved onto a Synergi Fusion RP column ($150mm{\times}2.0mm$, $4{\mu}m$) with a gradient elution of mobile phase A (0.1% formic acid in 2 mmol/L ammonium acetate, v/v) and mobile phase B (20% methanol in acetonitrile, v/v) at a flow rate of $450{\mu}L$/min. Porphyrins and the internal standard (IS), coproporphyrin I-$^{15}N_4$, were detected by a tandem mass spectrometer equipped with an electrospray ion source operating in positive ion mode. Multiple reaction monitoring (MRM) transitions of the protonated precursor ions and the related product ions were optimized to increase selectivity and sensitivity. The proposed method was validated by assessing selectivity, linearity, limit of quantification (LOQ), precision, accuracy, recovery, and stability. The calibration curves were obtained in the range of 0.1-100 nmol/L and the LOQs were estimated as 0.1 nmol/L for all porphyrins. Results obtained from the validation study of porphyrins showed good accuracy, precision, recovery, and stability. Finally, the proposed method was successfully applied to clinical studies on the autism spectrum disorder (ASD) diagnosis of 203 Korean children.

키워드

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피인용 문헌

  1. UHPLC–MS/MS bioanalysis of human plasma coproporphyrins as potential biomarkers for organic anion-transporting polypeptide-mediated drug interactions vol.10, pp.9, 2018, https://doi.org/10.4155/bio-2017-0246