title: Optimization and Evaluation Strategy of Esophageal Tissue Preparation Protocols for Metabolomics by LC-MS.
abstract: Sample preparation is a critical step in tissue metabolomics. Therefore, a comprehensive and systematic strategy for the screening of tissue preparation protocols is highly desirable. In this study, we developed an Optimization and Evaluation Strategy based on LC-MS to screen for a high-extractive efficiency and reproducible esophageal tissue preparation protocol for different types of endogenous metabolites (amino acids, carnitines, cholines, etc.), with a special focus on low-level metabolites. In this strategy, we first selected a large number of target metabolites based on literature survey, previous work in our lab, and known metabolic pathways. For these target metabolites, we tested different solvent extraction methods (biphasic solvent extraction, two-step [TS], stepwise [SW], all-in one [AO]; single-phase solvent extraction, SP) and esophageal tissue disruption methods (homogenized wet tissue [HW], ground wet tissue [GW], and ground dry tissue [GD]). A protocol involving stepwise addition of solvents and a homogenized wet tissue protocol (SWHW) was superior to the others. Finally, we evaluated the stability of endogenous metabolites in esophageal tissues and the sensitivity, reproducibility, and recovery of the optimal protocol. The results proved that the SWHW protocol was robust and adequate for bioanalysis. This strategy will provide important guidance for the standardized and scientific investigation of tissue metabolomics.
authors.affiliation: State Key Laboratory of Bioactive Substance and Function of Natural Medicines, Institute of Materia Medica, Chinese Academy of Medical Sciences and Peking Union Medical College , Beijing 100050, People's Republic of China.
authors.forename: Huiqing
authors.initials: H
authors.lastname: Wang
authors.affiliation: State Key Laboratory of Bioactive Substance and Function of Natural Medicines, Institute of Materia Medica, Chinese Academy of Medical Sciences and Peking Union Medical College , Beijing 100050, People's Republic of China.
authors.forename: Jing
authors.initials: J
authors.lastname: Xu
authors.affiliation: State Key Laboratory of Bioactive Substance and Function of Natural Medicines, Institute of Materia Medica, Chinese Academy of Medical Sciences and Peking Union Medical College , Beijing 100050, People's Republic of China.
authors.forename: Yanhua
authors.initials: Y
authors.lastname: Chen
authors.affiliation: State Key Laboratory of Bioactive Substance and Function of Natural Medicines, Institute of Materia Medica, Chinese Academy of Medical Sciences and Peking Union Medical College , Beijing 100050, People's Republic of China.
authors.forename: Ruiping
authors.initials: R
authors.lastname: Zhang
authors.affiliation: State Key Laboratory of Bioactive Substance and Function of Natural Medicines, Institute of Materia Medica, Chinese Academy of Medical Sciences and Peking Union Medical College , Beijing 100050, People's Republic of China.
authors.forename: Jiuming
authors.initials: J
authors.lastname: He
authors.affiliation: State Key Laboratory of Bioactive Substance and Function of Natural Medicines, Institute of Materia Medica, Chinese Academy of Medical Sciences and Peking Union Medical College , Beijing 100050, People's Republic of China.
authors.forename: Zhonghua
authors.initials: Z
authors.lastname: Wang
authors.affiliation: State Key Laboratory of Bioactive Substance and Function of Natural Medicines, Institute of Materia Medica, Chinese Academy of Medical Sciences and Peking Union Medical College , Beijing 100050, People's Republic of China.
authors.forename: Qingce
authors.initials: Q
authors.lastname: Zang
authors.affiliation: New Drug Safety Evaluation Center, Institute of Materia Medica, Peking Union Medical College , Beijing 100050, People's Republic of China.
authors.forename: Jinfeng
authors.initials: J
authors.lastname: Wei
authors.affiliation: State Key Laboratory of Bioactive Substance and Function of Natural Medicines, Institute of Materia Medica, Chinese Academy of Medical Sciences and Peking Union Medical College , Beijing 100050, People's Republic of China.
authors.forename: Xiaowei
authors.initials: X
authors.lastname: Song
authors.affiliation: State Key Laboratory of Bioactive Substance and Function of Natural Medicines, Institute of Materia Medica, Chinese Academy of Medical Sciences and Peking Union Medical College , Beijing 100050, People's Republic of China.
authors.forename: Zeper
authors.initials: Z
authors.lastname: Abliz
mesh_terms: D000818:Animals; D002851:Chromatography, High Pressure Liquid; D004947:Esophagus / Q000378:metabolism*; D013058:Mass Spectrometry; D055432:Metabolomics / Q000379:methods*; D051381:Rats