Application Note 55
Rapid Analysis of 17 Bile Acids in Human Plasma by LC-MS/MS
Haley Berkland1, Andrew J. Percy, PhD2
1. Restek Corporation, Bellefonte, PA USA
2. Cambridge Isotope Laboratories, Inc., Tewksbury, MA USA
Abstract
The quantitative analysis of bile acids in plasma is critical for diagnosing many structural liver diseases. Accurate reporting can be difficult because of analyte characteristics, matrix effects, and other factors. In this work, we describe a robust, selective LC-MS/MS method for the analysis of 17 bile acids in under 10 minutes, including three isomer groups, with special attention given to a matrix interference observed in routine human plasma analysis.
Introduction
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. There are two main types of bile acids based upon their functional groups: unconjugated (or free) and conjugated, primarily with glycine- or taurine-based residues (Figure 1, Table 1). Quantitation of bile acids in matrix can be very challenging due to several 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
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