Method for detecting bile acid in excrement by UPLC-MS/MS

A bile acid and feces technology, which is applied in the field of bile acid detection in feces, can solve the problems of insufficient detection range of bile acid, and achieve the effect of absolute quantification, strong specificity, and high sensitivity

Pending Publication Date: 2021-03-16
BEIJING NOVOGENE TECH CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0006] The main purpose of the present invention is to provide a method for UPLC-MS / MS combined detection of bile acids in feces to solve the problem of insufficient detection range of bile acids in feces in the prior art

Method used

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  • Method for detecting bile acid in excrement by UPLC-MS/MS
  • Method for detecting bile acid in excrement by UPLC-MS/MS
  • Method for detecting bile acid in excrement by UPLC-MS/MS

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0046] Internal standard working solution preparation:

[0047] Firstly, chenodeoxycholic acid-D4, cholic acid-D4, ursodeoxycholic acid-D4, glycochenodeoxycholic acid-D4, glycocholic acid-D4 and lithocholic acid- D4 was prepared into 1mg / mL internal standard mother solution respectively, and then the mother solution was diluted according to the concentration shown in Table 1 to obtain the internal standard stock solution, and the internal standard stock solution was processed in the next step according to the requirements in Table 1 to obtain the internal standard work liquid.

[0048] Table 1

[0049]

[0050] Chromatographic conditions:

[0051] Chromatographic column: Agela Venusil MP C18 2.1mm×100mm×2.5μm;

[0052] Mobile phase: A phase: 0.1% formic acid aqueous solution; B phase: acetonitrile;

[0053] Column temperature: 50°C;

[0054] Injection volume: 2μL;

[0055] The chromatographic gradient is shown in Table 2.

[0056] Table 2

[0057] time min...

Embodiment 2

[0082] Compared with Example 1, the difference is that the chromatographic gradient is as shown in Table 4.

[0083] Table 4

[0084] time min Flow rateml / min A％ B％ 0.0 0.5 80 20 0.5 0.5 80 20 2 0.5 65 35 10 0.5 50 50 16 0.5 0 100 16.1 0.5 80 20 19 0.5 80 20

[0085] According to the chromatographic gradient of Example 2, the mixed standard sample is separated, and the separation effect is worse than that of Example 1. The specific performance is that the peaks are unevenly distributed, and there are miscellaneous peaks at the end of the gradient. For details, see Figure 5 The total ion chromatogram is shown.

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Abstract

The invention provides a method for detecting bile acid in excrement by UPLC-MS/MS. The method comprises the following steps: establishing a standard curve; treating a to-be-detected excrement sample,and dissolving the frozen to-be-detected excrement sample by adopting mass spectrum water to obtain to-be-detected slurry; carrying out extraction and centrifugal separation on the to-be-detected slurry by adopting an extracting agent to obtain supernatant; drying the supernatant, and performing redissolving by adopting an initial mobile phase to obtain a redissolving system; centrifuging the redissolving system to obtain a to-be-detected solution, wherein the extracting agent comprises a mixed solution of acetonitrile and methanol, the volume ratio of acetonitrile to methanol in the mixed solution is 7:3-9:1, and the initial mobile phase comprises a mixed solution of acetonitrile and a 0.1% formic acid aqueous solution in a volume ratio of 7:3-9:1; detecting the to-be-detected solution by adopting UPLC-MS/MS to obtain ion spectrum peaks of bile acids; and converting the content of each bile acid according to the standard curve and the ion spectrum peak. The method can be used for quantifying 33 bile acids.

Description

technical field [0001] The invention relates to the technical field of detection of bile acids in feces, in particular to a method for detecting bile acids in feces by UPLC-MS / MS. Background technique [0002] Bile acid is an important component of bile. Its main physiological function is to maintain the balance of lipid metabolism in the body. In organisms, it usually uses cholesterol as a raw material to synthesize in the liver to generate primary bile acid. Primary bile acid is secreted with bile to the duodenum, and then through a series of reactions to generate secondary bile acids. In addition, primary bile acids are also combined with glycine or taurine to form conjugated bile acids. Bile acids will be reabsorbed by the intestine in the organism and taken up by the liver , so as to form bile for re-secretion. This process is enterohepatic circulation, which is repeated several times a day. During this process, 95% of bile acids are reabsorbed, and the other 5% will be...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N30/02G01N30/06G01N30/60G01N30/30G01N30/34G01N30/72
CPCG01N30/02G01N30/06G01N30/60G01N30/30G01N30/34G01N30/7266G01N2030/062G01N2030/045
Inventor 刘振东王劲松赵晓雯刘莹李朋朋
Owner BEIJING NOVOGENE TECH CO LTD
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