Abstract
A simple, sensitive, and specific liquid chromatography-electrospray tandem mass spectrometry (LC-MS/MS) method for the determination of bile acids in human bile has been developed. The bile acids were extracted with a C18 (octadecyl) reversed-phase column and identified and quantified by simultaneous monitoring of their parent and daughter ions, using the multiple reaction monitoring mode. Identification and quantification of conjugated bile acids in bile was achieved in 5 min. The detection limit was 1 ng, and the determination was linear for concentrations up to 100 ng. The percent recovery of standards made of single conjugated (glycine and taurine) bile acid or of mixture of glycine- or taurine-conjugated cholic acid, chenodeoxycholic acid, deoxycholic acid, ursodeoxycholic acid, and lithocholic acid averaged 71.73% to 95.92%. The percent recovery of the same standard bile acids was also determined by gas chromatography-mass spectrometry (GC-MS), using the selected ion monitoring mode, and averaged 66% to 96%. A biliary bile acid profile of human gallbladder bile was obtained by LC-MS/MS and GC-MS. The results showed a good correlation between the two techniques and no significant differences between the two methods were observed. The LC-MS/MS method was also used for the analysis of serum, urine, and fecal bile acids. In conclusion, LC-MS/MS is a simple, sensitive, and rapid technique for the analysis of conjugated bile acids in bile and other biological samples.—Perwaiz, S., B. Tuchweber, D. Mignault, T. Gilat, and I. M. Yousef. Determination of bile acids in biological fluids by liquid chromatography-electrospray tandem mass spectrometry. J. Lipid Res. 2001. 42: 114–119.
Highlights
A simple, sensitive, and specific liquid chromatography-electrospray tandem mass spectrometry (LC-MS/ MS) method for the determination of bile acids in human bile has been developed
Validation of the LC-MS/MS method The detection limit for the individual conjugated bile acids was 1 ng and the assay was linear over the range of 1–100 ng
We have developed a rapid, reproducible, accurate, and sensitive method, using LC-MS/MS, to determine conjugated and total bile acids in bile
Summary
A simple, sensitive, and specific liquid chromatography-electrospray tandem mass spectrometry (LC-MS/ MS) method for the determination of bile acids in human bile has been developed. The percent recovery of the same standard bile acids was determined by gas chromatography-mass spectrometry (GC-MS), using the selected ion monitoring mode, and averaged 66% to 96%. LC-MS/MS is a simple, sensitive, and rapid technique for the analysis of conjugated bile acids in bile and other biological samples.—Perwaiz, S., B. Determination of bile acids in biological fluids by liquid chromatography-electrospray tandem mass spectrometry. In the last decade liquid chromatography-mass spectrometry has been used for qualitative analysis of bile acids (14 –17). Bile acids are the major components of bile They are synthesized in the liver and secreted in the gallbladder or in the intestine, conjugated mainly with taurine and glycine.
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