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Determination of Bile Acids in Piglet Bile by Solid Phase Extraction and Liquid Chromatography-Electrospray Tandem Mass Spectrometry

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Lipids

Abstract

An LC/MS/MS-based method was developed for the determination of individual bile acids (BA) and their conjugates in porcine bile samples. The C18-based solid-phase extraction (SPE) procedure was optimized so that all 19 target BA and their glycine and taurine conjugates were collected with high recoveries for standards (89.1–100.2 %). Following this, all 19 compounds were separated and quantified in a single 12 min chromatographic run. The method was validated in terms of linearity, sensitivity, accuracy, precision, and recovery. An LOD in the low ppb range with measured precisions in the range of 0.5–9.3 % was achieved. The recoveries for all of the 19 analytes in bile samples were all >80 %. The validated method was successfully applied to the profiling of BA and their conjugates in the bile from piglets treated with exogenous glucagon-like peptide-2 (GLP-2) in a preclinical model of neonatal parenteral nutrition-associated liver disease (PNALD). The method developed is rapid and could be easily implemented for routine analysis of BA and their conjugates in other biofluids or tissues.

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Abbreviations

BA:

Bile acid

CA:

Cholic acid

CDCA:

Chenodeoxycholic acid

DCA:

Deoxycholic acid

ESI:

Electrospray

GCA-d4 :

Glycocholic acid-2,2,4,4-d4

GCDCA:

Glycochenodeoxycholic acid

GDCA:

Glycodeoxycholic acid

GHDCA:

Glycohyodeoxycholic acid

GLCA:

Glycolithocholic acid

GLP-2:

Glucagon-like peptide-2

GUDCA:

Glycoursodeoxycholic acid

HCA:

Hyocholic acid

HDCA:

Hyodeoxycholic acid

IS:

Internal standard

LCA:

Lithocholic acid

LC/MS/MS:

Liquid chromatography/tandem mass spectrometry

LOD:

Limit of detection

MCA:

Muricholic acid

MRM:

Multiple reaction monitoring

PNALD:

Parenteral nutrition-associated liver disease

SIR:

Selected ion recording

SPE:

Solid phase extraction

TCA:

Taurocholic acid

TCDCA:

Taurochenodeoxycholic acid

TMCA:

Tauromuricholic acid

TDCA:

Taurodeoxycholic acid

THCA:

Taurohyocholic acid

THDCA:

Taurohyodeoxycholic acid

TLCA:

Taurolithocholic acid

UDCA:

Ursodeoxycholic acid

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Acknowledgments

S. Mi acknowledges the generous support of a student award from the China Scholarship Council (CSC) and technical assistance from Dr. Y-Y. Zhao. D.W. Lim is supported by the University of Alberta Clinician-Investigator Program, the Canadian Institutes of Health Research (Frederick Banting and Charles Best Canada Graduate Scholarship—Doctoral), Alberta Innovates—Health Solutions (Clinician Fellowship), Killam Trusts (Honorary Izaak Walton Killam Memorial Doctoral Scholarship) and the Women’s and Children’s Health Research Institute (WCHRI, Graduate Studentship) through the generous support of the Stollery Children’s Hospital Foundation. J.M. Curtis acknowledges support from an NSERC Discovery grant.

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Authors and Affiliations

  1. Department of Agricultural, Food and Nutritional Science, University of Alberta, 3-60D South Academic Building, Edmonton, AB, T6G 2P5, Canada

    Si Mi & Jonathan M. Curtis

  2. Department of Surgery, University of Alberta, Edmonton, AB, Canada

    David W. Lim

  3. Department of Pediatrics, University of Alberta, Edmonton, AB, Canada

    Justine M. Turner

  4. Department of Surgery, Hospital for Sick Children and University of Toronto, Toronto, ON, Canada

    Paul W. Wales

Authors
  1. Si Mi
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  2. David W. Lim
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  3. Justine M. Turner
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  4. Paul W. Wales
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  5. Jonathan M. Curtis
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Corresponding author

Correspondence to Jonathan M. Curtis.

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Mi, S., Lim, D.W., Turner, J.M. et al. Determination of Bile Acids in Piglet Bile by Solid Phase Extraction and Liquid Chromatography-Electrospray Tandem Mass Spectrometry. Lipids 51, 359–372 (2016). https://doi.org/10.1007/s11745-016-4125-1

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  • DOI: https://doi.org/10.1007/s11745-016-4125-1

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