Simultaneous Absolute Quantification of 11 Cytochrome.pdf

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Simultaneous Absolute Quantification of 11 Cytochrome.pdf

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Simultaneous Absolute Quantification of 11 Cytochrome

Simultaneous Absolute Quantification of 11 Cytochrome P450 Isoforms in Human Liver Microsomes by Liquid Chromatography Tandem Mass Spectrometry with In Silico Target Peptide SelectionHIROTAKA KAWAKAMI,1 SUMIO OHTSUKI,1 JUNICHI KAMIIE,2 TAKASHI SUZUKI,3 TAKAAKI ABE,4 TETSUYA TERASAKI1 1Division of Membrane Transport and Drug Targeting, Graduate School of Pharmaceutical Sciences, Tohoku University, Aoba, Aramaki, Aoba-ku, Sendai 980-8578, Japan 2Laboratory of Veterinary Pathology, School of Veterinary Medicine, Azabu University, 1-17-71 Fuchinobe, Sagamihara, Kanagawa 229-8501, Japan 3Division of Pathology, Tohoku University Graduate School of Medicine, 2-1 Seiryou-machi, Aoba-ku, Sendai 980-8574, Japan 4Division of Nephrology, Endocrinology, and Vascular Medicine, Tohoku University Graduate School of Medicine, 2-1 Seiryou-machi, Aoba-ku, Sendai 980-8574, Japan Received 10 February 2010; revised 28 April 2010; accepted 3 May 2010 Published online 16 June 2010 in Wiley Online Library (). DOI 10.1002/jps.22255Abbreviation mass scan; EPI HLM, human liv tandem mass sp MS, mass spect Corresponden 6831; Fax: t81- E-mail: terasak Journal of Pharm 2010 Wiley-LissABSTRACT: Cytochrome P450 (CYP) proteins are involved in the biological oxidation and reduction of xenobiotics, affecting the pharmacological efficiency of drugs. This study aimed to establish a method to simultaneously quantify 11 CYP isoforms by multiplexed-multiple reaction monitoring analysis with liquid chromatography tandem mass spectrometry and in silico peptide selection to clarify CYP isoform expression profiles in human liver tissue. CYP1A2, 2A6, and 2D6 target peptides were identified by shot-gun proteomic analysis, and those of other isoforms were selected by in silico peptide selection criteria. The established quantification method detected target peptides at 10 fmol, and the dynamic range of calibration curves was at least 500-fold. The quantification value of CYP1A2 in Supersomes was not significantl