Detection method and application of metabolic profile of bile acid pathway based on serum samples

Abstract

The invention discloses a method for detecting the metabolic profile of bile acid pathways based on serum samples, comprising: preparing samples; using ultra-high performance liquid chromatography tandem time-of-flight mass spectrometry to deduce and identify bile acids in serum samples, and if there are standard products, use the standard products Confirm the retention time of the primary mass spectrometry quasi-molecular ion and the accurate mass-to-charge ratio of the characteristic secondary fragment ion; if there is no standard, according to the cracking law and chromatographic characteristics of the same type of bile acid, combined with the primary and secondary mass spectrometry in serum Identify the data and chromatographic behavior; use ultra-high performance liquid chromatography in series triple quadrupole mass spectrometry to quantitatively analyze bile acids, cholesterol and hydroxysterols. If there are standards, optimize the appropriate detection conditions through standards; Based on the qualitative first-order and second-order mass spectrometry data of ultra-high performance liquid chromatography tandem time-of-flight mass spectrometry, the characteristic product ions are optimized, the cone voltage and collision energy are optimized, and quantitative analysis is realized; the metabolic profile of the whole bile acid pathway is obtained.

Description

technical field [0001] The application belongs to the technical field of biological detection, and in particular relates to a detection method and application of a serum sample-based metabolic profile of the whole bile acid pathway. Background technique [0002] Bile acid is the end product of cholesterol metabolism in the liver and is the main way for the body to clear cholesterol. The human liver synthesizes about 200 to 600 mg of bile acids per day, which are actively transported into the bile and stored in the gallbladder through transporters such as the bile salt excretion pump (BSEP) and multidrug resistance protein 2 (MRP2). released into the duodenum. After the body eats, it is released into the duodenum under the stimulation of cholecystokinin (CCK). About 95% of the primary and secondary bile acid mixture in the intestinal tract enters the small intestinal mucosal cells through the bile acid transporter (ASBT), and enters the portal vein through the ileal bile ac...

AI Technical Summary

Problems solved by technology

[0005] However, free bile acids have a stable structure and are not easy to break into characteristic fragments; the characteristic fragment ions of bound bile acids come from the cleavage of shared side chain amido bonds, and the characteristic fragments have the same mass-to-charge ratio. It is difficult to separate parent and child ion pairs, which puts great demands on the separation ability of front-end liquid chromatography

In addition, due to the above-mentioned difficulties and the lack of standard products or the need for customization, there are few quantitative studies on the quantitative analysis of oxo- and sulfated bile acids in biological specimens, which hinders the evaluation of their potential physiological effects and the metabolic mechanism of the bile acid pathway. analyze

In the prior art, the currently developed LC-MS method can realize the detection of more than 40 kinds of endogenous bile acids in serum, and has not yet used LC-MS / MS to realize the detection of cholesterol, hydroxylated sterols, primary bile acids, and secondary bile acids. Detection method for the whole pathway metabolism of acid and tertiary bile acid

Images