Method for efficiently detecting 15 kinds of bile acid in blood

A bile acid and blood technology, which is applied in the field of efficient detection of 15 kinds of bile acids in blood, can solve the problems of difficult bile acid analysis and measurement, biosafety problems, cross-contamination, etc., and achieves strong specificity, strong stability and reliability sex high effect

Inactive Publication Date: 2017-11-17
HANGZHOU HEALTH BANK MEDICAL LAB CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, it is difficult to accurately analyze and measure the contents of different bile acids in the blood by the conventional methods adopted in the current prior art
[0005] At the same time, in the traditional bile acid detection method, due to the long-distance transportation of blood samples, it is not only inconvenient to store, but also prone to cross-contamination and lead to corresponding biosafety problems.

Method used

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  • Method for efficiently detecting 15 kinds of bile acid in blood
  • Method for efficiently detecting 15 kinds of bile acid in blood
  • Method for efficiently detecting 15 kinds of bile acid in blood

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0048] The detection of 15 kinds of bile acids in embodiment 1 human blood

[0049] 1 material

[0050] 1.1 Instruments and materials

[0051] American AB Sciex tandem mass spectrometer, Angilent 1290 liquid chromatograph; Agilent C8 column (2.1×50mm, 1.7μm).

[0052] 1.2 Drugs and reagents

[0053] Standard substances: cholic acid (CA), ursodeoxycholic acid (UDCA), chenodeoxycholic acid (CDCA), deoxycholic acid (DCA), lithocholic acid (LCA), glycocholic acid sodium salt (GCA), glycocholate Ursodeoxycholic acid (GUDCA), glycochenodeoxycholic acid sodium salt (GCDCA), glycodeoxycholic acid sodium salt (GDCA), taurocholic acid sodium salt (TDCA), taurodeoxycholic acid sodium salt ( TCA), tauroursodeoxycholic acid sodium salt (TUDCA), taurochenodeoxycholic acid sodium salt (TCDCA), glycinyllithocholic acid (GLCA), taurolithocholic acid sodium salt (TLCA) were purchased from Sigma company.

[0054] The isotope internal standard of the above bile acids is deoxycholic acid DCA~...

experiment example 1

[0088] The recovery rate analysis of experimental example 1 inventive method

[0089] Take 200 μL each of the standard solutions of high, medium and low concentrations, and make up to 1ml with methanol solution, make it fully mixed, and directly take 10-200 μL of the supernatant and drop it on the filter paper, air-dry for more than 3 hours, and use it with bile acid isotope The internal standard is extracted with 0.01-10% formic acid methanol or 0.01-10% formic acid-acetonitrile mixed solution, and mixed with ultrasonic / constant temperature oscillation / hybridization oven, and then centrifuged at 10000-18000rpm for 10min to take the supernatant, freeze-dried / dried with nitrogen; Add 50-300 μL of 10-80% methanol water to redissolve, vortex, centrifuge at 10000-18000 rpm for 10 min, and take the supernatant for LC-MS analysis. The liquid chromatography and mass spectrometry conditions of the liquid chromatography-mass spectrometry in the detection process are all the same as the...

experiment example 2

[0096] Experimental example 2 practical application cases of the inventive method

[0097] 95 samples were taken, including 30 healthy neonates, 25 physiological jaundice neonates, and 40 pathological jaundice neonates. When processing the sample, drop 10-200 μL of whole blood onto a filter paper sheet and air-dry at room temperature for >3 hours; punch a hole in the air-dried dried blood sheet with a pore size of 3 mm, take a certain number of sheets and place them in a 1.5 mL centrifuge tube, and use The isotopic internal standard is extracted with 0.01-10% formic acid methanol or 0.01-10% formic acid-acetonitrile mixed solution, and mixed with ultrasonic / constant temperature oscillation / hybridization oven, and then centrifuged at 10000-18000rpm for 10min to take the supernatant, freeze-dried / nitrogen blow-dried ; Add 50-300 μL of 10-80% methanol water to redissolve, vortex, centrifuge at 10000-18000 rpm for 10 min, and take the supernatant for LC-MS analysis. The ratio of ...

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Abstract

The invention provides a method for efficiently detecting 15 kinds of bile acid in blood. In the method, serum is directly preserved on a filter paper scrip, and through an efficient extraction process, extracted bile acid is directly injected on a liquid chromatography-mass spectroscopy; 15 kinds of the bile acid in blood can be efficiently detected in a high-throughput and high-repeatability mode, and the method has the wide application value.

Description

technical field [0001] The invention relates to the field of bile acid detection, in particular to a method for efficiently detecting 15 kinds of bile acids in blood. Background technique [0002] Bile acid is the main organic component in bile, which contains a variety of steroid compounds with similar structures. Bile acids have important physiological functions such as emulsifying fat, producing bile, and preventing gallstones in the body. [0003] Current studies have found that a variety of hepatobiliary diseases can cause bile acid enterohepatic circulation disorders, resulting in abnormal serum bile acid profiles, and this abnormality will also affect the synthesis and secretion of bile acids in the liver. Therefore, serum bile acid is an important marker to measure whether the liver and gallbladder function is normal or not. [0004] In traditional clinical practice, only total bile levels are generally measured. However, because the toxicity of different bile aci...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N30/02G01N30/06
CPCG01N30/02G01N30/06G01N2030/062
Inventor 童鸿斌朱雅玲叶剑
Owner HANGZHOU HEALTH BANK MEDICAL LAB CO LTD
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