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

Yudai Suzuki, Rina Kaneko, Mina Nomura, Hisao Naito, Kazuya Kitamori, Tamie Nakajima, Tadashi Ogawa, Hideki Hattori, Hiroshi Seno, Akira Ishii

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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 μg/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).

Original languageEnglish
Pages (from-to)57-71
Number of pages15
JournalNagoya journal of medical science
Volume76
Issue number1-2
Publication statusPublished - 20-03-2014

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High Fat Diet
Bile Acids and Salts
Glycine
Liver
Serum
Cholesterol
Diet
Taurine
Inbred SHR Rats
Tandem Mass Spectrometry
Liquid Chromatography
Calibration
Limit of Detection
Stroke
Fats
Acids

All Science Journal Classification (ASJC) codes

  • Medicine(all)

Cite this

Suzuki, Yudai ; Kaneko, Rina ; Nomura, Mina ; Naito, Hisao ; Kitamori, Kazuya ; Nakajima, Tamie ; Ogawa, Tadashi ; Hattori, Hideki ; Seno, Hiroshi ; Ishii, Akira. / 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. In: Nagoya journal of medical science. 2014 ; Vol. 76, No. 1-2. pp. 57-71.
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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 μg/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).",
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Suzuki, Y, Kaneko, R, Nomura, M, Naito, H, Kitamori, K, Nakajima, T, Ogawa, T, Hattori, H, Seno, H & Ishii, A 2014, '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', Nagoya journal of medical science, vol. 76, no. 1-2, pp. 57-71.

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. / Suzuki, Yudai; Kaneko, Rina; Nomura, Mina; Naito, Hisao; Kitamori, Kazuya; Nakajima, Tamie; Ogawa, Tadashi; Hattori, Hideki; Seno, Hiroshi; Ishii, Akira.

In: Nagoya journal of medical science, Vol. 76, No. 1-2, 20.03.2014, p. 57-71.

Research output: Contribution to journalArticle

TY - JOUR

T1 - Simple and rapid quantitation of 21 bile acids in rat serum and liver by UPLC-MS-MS

T2 - Effect of high fat diet on glycine conjugates of rat bile acids

AU - Suzuki, Yudai

AU - Kaneko, Rina

AU - Nomura, Mina

AU - Naito, Hisao

AU - Kitamori, Kazuya

AU - Nakajima, Tamie

AU - Ogawa, Tadashi

AU - Hattori, Hideki

AU - Seno, Hiroshi

AU - Ishii, Akira

PY - 2014/3/20

Y1 - 2014/3/20

N2 - 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 μg/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).

AB - 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 μg/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).

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