title: Simple and rapid quantitation of 21 bile acids in rat serum and liver by UPLC-MS-MS: effect of high fat diet on glycine conjugates of rat bile acids.
abstract: In this report, we present a simple and rapid method for analysis of 21 kinds of bile acids and the conjugates in rat serum and liver samples by ultra-performance liquid chromatography-tandem mass spectrometry (UPLC-MS-MS) in the negative ionization mode, using cholic-2, 2, 4, 4-d4 acid as internal standard. After liquid-liguid extraction from serum and liver samples, specimens were analyzed by UPLC equipped with an Acquity TQD tandem quadrupole mass spectrometer. All of the 21 bile acids were sufficiently separated within 5 min. For most bile acids, calibration curves showed good linearities in the range of 0.25 to 5000 ng/mL for serum samples, 2.5 ng/g to 50 microg/g for liver samples. The limits of detection (LOD) were estimated to be less than 0.25 to 7.5 ng/mL in serum, less than 2.5 to 10 ng/g in liver samples. The present method was validated with respect to repeatability; the coefficient of variation (CV) values were less than 26.7% in the serum and 25.9% in the liver. In the animal study, we compared 21 bile acids in the serum and liver samples of the stroke-prone spontaneously hypertensive (SHRSP) rats fed with control (SP) diet or high-fat and high-cholesterol-containing (HFC) diet. By feeding with HFC diet, the glycine conjugates of some bile acids significantly increased and the taurine conjugate of ulsodeoxicolate (TUDC) decreased in serum and liver samples. Our results suggest that the change of bile acid profiles could be applied for the diagnosis of non-alcoholic fatty liver disease (NAFLD).
authors.affiliation: Department of Legal Medicine and Bioethics, Nagoya University Graduate School of Medicine, Nagoya, Japan.
authors.forename: Yudai
authors.initials: Y
authors.lastname: Suzuki
authors.forename: Rina
authors.initials: R
authors.lastname: Kaneko
authors.forename: Mina
authors.initials: M
authors.lastname: Nomura
authors.forename: Hisao
authors.initials: H
authors.lastname: Naito
authors.forename: Kazuya
authors.initials: K
authors.lastname: Kitamori
authors.forename: Tamie
authors.initials: T
authors.lastname: Nakajima
authors.forename: Tadashi
authors.initials: T
authors.lastname: Ogawa
authors.forename: Hideki
authors.initials: H
authors.lastname: Hattori
authors.forename: Hiroshi
authors.initials: H
authors.lastname: Seno
authors.forename: Akira
authors.initials: A
authors.lastname: Ishii
mesh_terms: D000818:Animals; D001647:Bile Acids and Salts / Q000097:blood*; D015415:Biomarkers / Q000097:blood; D002138:Calibration; D002791:Cholesterol, Dietary / Q000097:blood*; D002853:Chromatography, Liquid / Q000379:methods* / Q000592:standards; D059305:Diet, High-Fat; D004195:Disease Models, Animal; D005234:Fatty Liver / Q000097:blood* / Q000175:diagnosis / Q000209:etiology; D005998:Glycine / Q000031:analogs & derivatives / Q000097:blood*; D057230:Limit of Detection; D016014:Linear Models; D059625:Liquid-Liquid Extraction; D008099:Liver / Q000378:metabolism* / Q000473:pathology; D065626:Non-alcoholic Fatty Liver Disease; D011237:Predictive Value of Tests; D051381:Rats; D011918:Rats, Inbred SHR; D012015:Reference Standards; D015203:Reproducibility of Results; D053719:Tandem Mass Spectrometry / Q000592:standards; D013654:Taurine / Q000097:blood; D013997:Time Factors
chemical_list: D001647:Bile Acids and Salts; D015415:Biomarkers; D002791:Cholesterol, Dietary; D013654:Taurine; D005998:Glycine
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