Quantitative analysis of bile acids in plasma is critical for diagnosing liver disease as well as assessing drug safety. Accurate reporting can be difficult because of analyte characteristics, matrix effects, and other factors. Here, we establish a rapid, robust, and selective LC-MS/MS method for the analysis of bile acids in human plasma using a Restek Raptor C18 column. One of the significant improvements over other reported methods was the baseline separation of all 17 bile acids in 6 minutes, including three isomer groups.
Bile acids are a group of major catabolic products of cholesterol, and they are critical signaling molecules that regulate cholesterol and glucose. The analysis of bile acids in human plasma is an important diagnostic tool as bile acids are biomarkers of liver disease and are also used as indicators of potentially harmful side effects of new drugs.
Quantitation of bile acids in matrix can be very challenging due to a number of factors. These include structural similarities, varying polarity and stereochemistry, the presence of isomers, limited fragmentation of unconjugated bile acids in a mass spectrometer, high endogenous levels, and matrix effects caused by phospholipids or triglycerides.
Traditionally, the analysis of bile acids in human plasma takes from 20 to over 60 min and results in only minimum resolution of isomers. Now, a simple UHPLC method was developed for the simultaneous quantification of the major bile acids, as well as
their glycine and taurine conjugates in human plasma. Using a Raptor C18 column and the methodology established here, baseline resolution and good chromatographic and quantitative results were obtained for all compounds in a fast, 6-min separation with a total analysis time of only 8.5 min.
From Restek: Lit. Cat.# CFAN2911-UNV
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