Development and Validation of a Highly Sensitive LC-MS/MS Method for the Analysis of Bile Acids in Serum, Plasma, and Liver Tissue Samples

Abstract

<jats:p>Bile acids control lipid homeostasis by regulating uptake from food and excretion. Additionally, bile acids are bioactive molecules acting through receptors and modulating various physiological processes. Impaired bile acid homeostasis is associated with several diseases and drug-induced liver injury. Individual bile acids may serve as disease and drug toxicity biomarkers, with a great demand for improved bile acid quantification methods. We developed, optimized, and validated an LC-MS/MS method for quantification of 36 bile acids in serum, plasma, and liver tissue samples. The simultaneous quantification of important free and taurine- and glycine-conjugated bile acids of human and rodent species has been achieved using a simple workflow. The method was applied to a mouse model of statin-induced myotoxicity to assess a possible role of bile acids. Treatment of mice for three weeks with 5, 10, and 25 mg/kg/d simvastatin, causing adverse skeletal muscle effects, did not alter plasma and liver tissue bile acid profiles, indicating that bile acids are not involved in statin-induced myotoxicity. In conclusion, the established LC-MS/MS method enables uncomplicated sample preparation and quantification of key bile acids in serum, plasma, and liver tissue of human and rodent species to facilitate future studies of disease mechanisms and drug-induced liver injury.</jats:p>

Impact on Bile Acid Concentrations by Alveolar Echinococcosis and Treatment with Albendazole in Mice

“…To assess the potential effects of E. multilocularis infection and its main treatment ABZ on the serum bile acid profiles in mice, a recently established LC-MS/MS-based method was applied to quantify a series of conjugated and unconjugated bile acids [ 28 ]. Glycine-conjugated bile acids were at or below the lower limit of detection of the quantification method and are not listed in Table 1 .…”

“…In addition, bile acids were quantified in the liver tissue using the previously described LC-MS/MS method [ 28 ]. The mean concentrations of each individual bile acid in addition to the sums of unconjugated, conjugated, and total bile acids analyzed for the four treatment groups are listed in Table 2 , and the data for individual bile acids are depicted in Supplementary Figure S2 .…”

“…≥98%) from Sigma-Aldrich (St. Louis, MO, USA). Bile acids and internal standards were purchased from Sigma-Aldrich or Steraloids (Newport, RI, USA) as described recently [ 28 ].…”

“…Bile acids were quantified as described recently [ 28 ]. Briefly, 10 µL of serum were diluted 1:4 with water, followed by adding 900 µL of 2-propanol for protein precipitation and a mixture of deuterated internal standards.…”

Using nontargeted LC-MS metabolomics to identify the association of biomarkers in pig feces with feed efficiency

“…The speci c synthesis process is that cholesterol 7α-hydroxylase (CYP27A1) catalyzes the oxidation of steroid side chains to form THC26 or DHCA in the mitochondria of liver cells and then obtains the primary bile acid cholic acid (CD) or chenodeoxycholic acid (CDCA) under the catalysis of various enzymes [19,[29][30][31][32]. Interestingly, although the synthesis of bile acids is determined by a variety of cytochrome P450 enzymes (CYPs), both THC26 and DHCA are intermediate products catalyzed by CYP27A1 [33]. Bile acids start from the catabolism of cholesterol and are the nal product of cholesterol catabolism; they play a critical role in food digestion and nutrient absorption, helping the absorption of lipids and fat-soluble vitamins in the intestine [32,[34][35][36].…”