title: An UPLC-MS/MS method for quantitative profiling of bile acids in sea lamprey plasma and tissues.
abstract: Bile acids (BAs) have recently gained more attention because of their diverse roles from digestion to signaling. Simultaneous analyses of various BAs in biological samples are challenging due to their structural similarity, relatively low concentrations, and the presence of isomeric forms. In this study, we report a simple and sensitive UPLC-MS/MS method for simultaneous quantifications of 13 BAs including four unique sea lamprey BAs in sea lamprey plasma, liver, intestine, and gills. A straightforward protein precipitation (PPT) method was used to extract BAs from the biological samples. Separation of all target analytes was achieved on a reverse-phase UPLC column in 15min, and detection was carried out on MS/MS with ESI in the negative ionization mode. This method was validated regarding its linearity, limits of detection (LOD), recovery, matrix effect, reproducibility, accuracy and precision. Significant improvements compared to previous LC-MS/MS methods were observed as a result of the application of UPLC and extensive optimization of experimental conditions. The method showed excellent linearity with high regression coefficients (>0.99) over a range of 0.5-1000ng/mL and LODs ranged from 0.009 to 0.11ng/mL. The applications of the developed method demonstrated that it simultaneously determined all target BAs in different biological sample matrices with excellent sensitivity, selectivity and reproducibility.
authors.affiliation: Department of Fisheries and Wildlife, Michigan State University, Room 13, Natural Resources Building, 480 Wilson Road, East Lansing, MI 48824, USA. Electronic address: hywang@msu.edu.
authors.forename: Huiyong
authors.initials: H
authors.lastname: Wang
authors.affiliation: Department of Fisheries and Wildlife, Michigan State University, Room 13, Natural Resources Building, 480 Wilson Road, East Lansing, MI 48824, USA.
authors.forename: Chu-Yin
authors.initials: CY
authors.lastname: Yeh
authors.affiliation: Department of Fisheries and Wildlife, Michigan State University, Room 13, Natural Resources Building, 480 Wilson Road, East Lansing, MI 48824, USA.
authors.forename: Ke
authors.initials: K
authors.lastname: Li
authors.affiliation: Department of Fisheries and Wildlife, Michigan State University, Room 13, Natural Resources Building, 480 Wilson Road, East Lansing, MI 48824, USA.
authors.forename: Yu-Wen
authors.initials: YW
authors.lastname: Chung-Davidson
authors.affiliation: Department of Fisheries and Wildlife, Michigan State University, Room 13, Natural Resources Building, 480 Wilson Road, East Lansing, MI 48824, USA. Electronic address: liweim@msu.edu.
authors.forename: Weiming
authors.initials: W
authors.lastname: Li
mesh_terms: D000818:Animals; D001647:Bile Acids and Salts / Q000097:blood* / Q000378:metabolism*; D048428:Petromyzon / Q000097:blood / Q000378:metabolism; D015203:Reproducibility of Results; D053719:Tandem Mass Spectrometry / Q000379:methods*
chemical_list: D001647:Bile Acids and Salts
keywords: Bile acids; Plasma; Protein precipitation; Sea lamprey; Tissue; UPLC–MS/MS