title: High-Throughput and Reliable Isotope Label-free Approach for Profiling 24 Metabolic Enzymes in FVB Mice and Sex Differences.
abstract: FVB mice are extensively used in transgenic and pharmacokinetic research. In this study, a validated isotope label-free method was constructed using ultrahigh-performance liquid chromatography-tandem mass spectrometry (UHPLC-MS/MS) to quantify 24 drug-metabolizing enzymes (DMEs) in FVB mice. The DMEs include cytochrome P450s (CYP450s/Cyp450s), UDP-glucuronsyltransferases (UGTs/Ugts), and sulfotransferases (SULTs/Sults), which catalyze a variety of reactions to detoxify xenobiotics and endobiotics. The proposed UHPLC-MS/MS method exhibited good range and high sensitivity for signature peptides, as well as acceptable accuracy, precision, and recovery. The protein expression profiles of the DMEs were determined in male and female mice. Overall, the major Cyps, Ugts, and Sults were expressed in male mice followed the rank order: Cyp2c29 > 2e1 > 3a11 > 1a2 > 2d22 > 27a1 > 2c39; Ugt2b5 > 2b1 > 1a6a > 1a9 > 1a1 > 2a3 > 1a2 > 1a5; and Sult1a1 > 3a > 1d1. In contrast, the rank order in female mice was Cyp2c29 > 2e1 > 2c39 > 2d22 > 3a11 > 1a2 > 27a1; Ugt1a6a > 2b5 > 1a1 > 2b1 > 2a3 > 1a9 > 1a5 > 1a2; and Sult1a1 > 3a1 > 1d1. Cyp2c29, Cyp1a2, Cyp27a1, Ugt2b1, Ugt2b5 and Ugt2b36 were male predominant, whereas Cyp2c39, Cyp2d22, Cyp7a1, Ugt1a1, Ugt1a5, Sult1a1, Sult3a1, and Sult1d1 were female predominant. This work could serve as a useful reference for the metabolic study of new drugs and for elucidating the effectiveness and toxicity of drugs. The method is stable, simple, and rapid for determining the expression of DMEs in animals.
authors.affiliation: International Institute for Translational Chinese Medicine, Guangzhou University of Chinese Medicine, Guangzhou, Guangdong, China (J.C., L.Z., X.L., H.Z., S.Z., J.Y., H.J., L.L., X.Q., Y.W., M.H., Z.L.); State Key Laboratory of Quality Research in Chinese Medicine, Macau University of Science and Technology, Macau, China (T.Y.); Department of Pharmacological and Pharmaceutical Sciences, College of Pharmacy, University of Houston, Houston, Texas (M.H.); and Department of Pharmaceutics, School of Pharmaceutical Sciences, Southern Medical University, Guangzhou, Guangdong, China (C.X.).
authors.forename: Jiamei
authors.initials: J
authors.lastname: Chen
authors.affiliation: International Institute for Translational Chinese Medicine, Guangzhou University of Chinese Medicine, Guangzhou, Guangdong, China (J.C., L.Z., X.L., H.Z., S.Z., J.Y., H.J., L.L., X.Q., Y.W., M.H., Z.L.); State Key Laboratory of Quality Research in Chinese Medicine, Macau University of Science and Technology, Macau, China (T.Y.); Department of Pharmacological and Pharmaceutical Sciences, College of Pharmacy, University of Houston, Houston, Texas (M.H.); and Department of Pharmaceutics, School of Pharmaceutical Sciences, Southern Medical University, Guangzhou, Guangdong, China (C.X.).
authors.forename: Lijun
authors.initials: L
authors.lastname: Zhu
authors.affiliation: International Institute for Translational Chinese Medicine, Guangzhou University of Chinese Medicine, Guangzhou, Guangdong, China (J.C., L.Z., X.L., H.Z., S.Z., J.Y., H.J., L.L., X.Q., Y.W., M.H., Z.L.); State Key Laboratory of Quality Research in Chinese Medicine, Macau University of Science and Technology, Macau, China (T.Y.); Department of Pharmacological and Pharmaceutical Sciences, College of Pharmacy, University of Houston, Houston, Texas (M.H.); and Department of Pharmaceutics, School of Pharmaceutical Sciences, Southern Medical University, Guangzhou, Guangdong, China (C.X.).
authors.forename: Xiaoyan
authors.initials: X
authors.lastname: Li
authors.affiliation: International Institute for Translational Chinese Medicine, Guangzhou University of Chinese Medicine, Guangzhou, Guangdong, China (J.C., L.Z., X.L., H.Z., S.Z., J.Y., H.J., L.L., X.Q., Y.W., M.H., Z.L.); State Key Laboratory of Quality Research in Chinese Medicine, Macau University of Science and Technology, Macau, China (T.Y.); Department of Pharmacological and Pharmaceutical Sciences, College of Pharmacy, University of Houston, Houston, Texas (M.H.); and Department of Pharmaceutics, School of Pharmaceutical Sciences, Southern Medical University, Guangzhou, Guangdong, China (C.X.).
authors.forename: Haihui
authors.initials: H
authors.lastname: Zheng
authors.affiliation: International Institute for Translational Chinese Medicine, Guangzhou University of Chinese Medicine, Guangzhou, Guangdong, China (J.C., L.Z., X.L., H.Z., S.Z., J.Y., H.J., L.L., X.Q., Y.W., M.H., Z.L.); State Key Laboratory of Quality Research in Chinese Medicine, Macau University of Science and Technology, Macau, China (T.Y.); Department of Pharmacological and Pharmaceutical Sciences, College of Pharmacy, University of Houston, Houston, Texas (M.H.); and Department of Pharmaceutics, School of Pharmaceutical Sciences, Southern Medical University, Guangzhou, Guangdong, China (C.X.).
authors.forename: Tongmeng
authors.initials: T
authors.lastname: Yan
authors.affiliation: International Institute for Translational Chinese Medicine, Guangzhou University of Chinese Medicine, Guangzhou, Guangdong, China (J.C., L.Z., X.L., H.Z., S.Z., J.Y., H.J., L.L., X.Q., Y.W., M.H., Z.L.); State Key Laboratory of Quality Research in Chinese Medicine, Macau University of Science and Technology, Macau, China (T.Y.); Department of Pharmacological and Pharmaceutical Sciences, College of Pharmacy, University of Houston, Houston, Texas (M.H.); and Department of Pharmaceutics, School of Pharmaceutical Sciences, Southern Medical University, Guangzhou, Guangdong, China (C.X.).
authors.forename: Cong
authors.initials: C
authors.lastname: Xie
authors.affiliation: International Institute for Translational Chinese Medicine, Guangzhou University of Chinese Medicine, Guangzhou, Guangdong, China (J.C., L.Z., X.L., H.Z., S.Z., J.Y., H.J., L.L., X.Q., Y.W., M.H., Z.L.); State Key Laboratory of Quality Research in Chinese Medicine, Macau University of Science and Technology, Macau, China (T.Y.); Department of Pharmacological and Pharmaceutical Sciences, College of Pharmacy, University of Houston, Houston, Texas (M.H.); and Department of Pharmaceutics, School of Pharmaceutical Sciences, Southern Medical University, Guangzhou, Guangdong, China (C.X.).
authors.forename: Sijing
authors.initials: S
authors.lastname: Zeng
authors.affiliation: International Institute for Translational Chinese Medicine, Guangzhou University of Chinese Medicine, Guangzhou, Guangdong, China (J.C., L.Z., X.L., H.Z., S.Z., J.Y., H.J., L.L., X.Q., Y.W., M.H., Z.L.); State Key Laboratory of Quality Research in Chinese Medicine, Macau University of Science and Technology, Macau, China (T.Y.); Department of Pharmacological and Pharmaceutical Sciences, College of Pharmacy, University of Houston, Houston, Texas (M.H.); and Department of Pharmaceutics, School of Pharmaceutical Sciences, Southern Medical University, Guangzhou, Guangdong, China (C.X.).
authors.forename: Jia
authors.initials: J
authors.lastname: Yu
authors.affiliation: International Institute for Translational Chinese Medicine, Guangzhou University of Chinese Medicine, Guangzhou, Guangdong, China (J.C., L.Z., X.L., H.Z., S.Z., J.Y., H.J., L.L., X.Q., Y.W., M.H., Z.L.); State Key Laboratory of Quality Research in Chinese Medicine, Macau University of Science and Technology, Macau, China (T.Y.); Department of Pharmacological and Pharmaceutical Sciences, College of Pharmacy, University of Houston, Houston, Texas (M.H.); and Department of Pharmaceutics, School of Pharmaceutical Sciences, Southern Medical University, Guangzhou, Guangdong, China (C.X.).
authors.forename: Huangyu
authors.initials: H
authors.lastname: Jiang
authors.affiliation: International Institute for Translational Chinese Medicine, Guangzhou University of Chinese Medicine, Guangzhou, Guangdong, China (J.C., L.Z., X.L., H.Z., S.Z., J.Y., H.J., L.L., X.Q., Y.W., M.H., Z.L.); State Key Laboratory of Quality Research in Chinese Medicine, Macau University of Science and Technology, Macau, China (T.Y.); Department of Pharmacological and Pharmaceutical Sciences, College of Pharmacy, University of Houston, Houston, Texas (M.H.); and Department of Pharmaceutics, School of Pharmaceutical Sciences, Southern Medical University, Guangzhou, Guangdong, China (C.X.).
authors.forename: Linlin
authors.initials: L
authors.lastname: Lu
authors.affiliation: International Institute for Translational Chinese Medicine, Guangzhou University of Chinese Medicine, Guangzhou, Guangdong, China (J.C., L.Z., X.L., H.Z., S.Z., J.Y., H.J., L.L., X.Q., Y.W., M.H., Z.L.); State Key Laboratory of Quality Research in Chinese Medicine, Macau University of Science and Technology, Macau, China (T.Y.); Department of Pharmacological and Pharmaceutical Sciences, College of Pharmacy, University of Houston, Houston, Texas (M.H.); and Department of Pharmaceutics, School of Pharmaceutical Sciences, Southern Medical University, Guangzhou, Guangdong, China (C.X.).
authors.forename: Xiaoxiao
authors.initials: X
authors.lastname: Qi
authors.affiliation: International Institute for Translational Chinese Medicine, Guangzhou University of Chinese Medicine, Guangzhou, Guangdong, China (J.C., L.Z., X.L., H.Z., S.Z., J.Y., H.J., L.L., X.Q., Y.W., M.H., Z.L.); State Key Laboratory of Quality Research in Chinese Medicine, Macau University of Science and Technology, Macau, China (T.Y.); Department of Pharmacological and Pharmaceutical Sciences, College of Pharmacy, University of Houston, Houston, Texas (M.H.); and Department of Pharmaceutics, School of Pharmaceutical Sciences, Southern Medical University, Guangzhou, Guangdong, China (C.X.).
authors.forename: Ying
authors.initials: Y
authors.lastname: Wang
authors.affiliation: International Institute for Translational Chinese Medicine, Guangzhou University of Chinese Medicine, Guangzhou, Guangdong, China (J.C., L.Z., X.L., H.Z., S.Z., J.Y., H.J., L.L., X.Q., Y.W., M.H., Z.L.); State Key Laboratory of Quality Research in Chinese Medicine, Macau University of Science and Technology, Macau, China (T.Y.); Department of Pharmacological and Pharmaceutical Sciences, College of Pharmacy, University of Houston, Houston, Texas (M.H.); and Department of Pharmaceutics, School of Pharmaceutical Sciences, Southern Medical University, Guangzhou, Guangdong, China (C.X.).
authors.forename: Ming
authors.initials: M
authors.lastname: Hu
authors.affiliation: International Institute for Translational Chinese Medicine, Guangzhou University of Chinese Medicine, Guangzhou, Guangdong, China (J.C., L.Z., X.L., H.Z., S.Z., J.Y., H.J., L.L., X.Q., Y.W., M.H., Z.L.); State Key Laboratory of Quality Research in Chinese Medicine, Macau University of Science and Technology, Macau, China (T.Y.); Department of Pharmacological and Pharmaceutical Sciences, College of Pharmacy, University of Houston, Houston, Texas (M.H.); and Department of Pharmaceutics, School of Pharmaceutical Sciences, Southern Medical University, Guangzhou, Guangdong, China (C.X.) liuzq@gzucm.edu.cn.
authors.forename: Zhongqiu
authors.initials: Z
authors.lastname: Liu
mesh_terms: D000818:Animals; D002851:Chromatography, High Pressure Liquid / Q000379:methods; D003577:Cytochrome P-450 Enzyme System / Q000378:metabolism; D004798:Enzymes / Q000378:metabolism*; D005260:Female; D014453:Glucuronosyltransferase / Q000378:metabolism; D007554:Isotopes / Q000378:metabolism*; D008297:Male; D055442:Metabolome / Q000502:physiology*; D055432:Metabolomics / Q000379:methods; D051379:Mice; D008815:Mice, Inbred Strains; D012727:Sex Characteristics; D015238:Sulfotransferases / Q000378:metabolism; D053719:Tandem Mass Spectrometry / Q000379:methods
chemical_list: D004798:Enzymes; D007554:Isotopes; D003577:Cytochrome P-450 Enzyme System; D014453:Glucuronosyltransferase; D015238:Sulfotransferases