Method for detecting five free bile acids in serum through high performance liquid chromatography-tandem mass spectrum

A high-performance liquid chromatography and tandem mass spectrometry technology, which is applied in the field of high-performance liquid chromatography-tandem mass spectrometry detection of five free bile acids, can solve the problems of complicated operation, insufficient sensitivity and specificity in the detection of free bile acids, and achieves the removal of serum Matrix interference, simple pretreatment steps, and good specificity

Inactive Publication Date: 2017-06-13
HANGZHOU BAICHEN MEDICAL LAB CO LTD
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Problems solved by technology

[0005] In order to overcome the deficiencies in the prior art, the object of the present invention is to provide a method for detecting five kinds of free bile acids in serum by high perfor...

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  • Method for detecting five free bile acids in serum through high performance liquid chromatography-tandem mass spectrum
  • Method for detecting five free bile acids in serum through high performance liquid chromatography-tandem mass spectrum
  • Method for detecting five free bile acids in serum through high performance liquid chromatography-tandem mass spectrum

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Embodiment Construction

[0039] Below, in conjunction with accompanying drawing and specific embodiment, the present invention is described further:

[0040] Five free bile acids: cholic acid, lithocholic acid, deoxycholic acid, ursodeoxycholic acid, chenodeoxycholic acid standard and isotope-labeled internal standard: deuterated cholic acid, deuterated deoxycholic acid, deuterated ursodeoxycholic acid Deoxycholic acid was purchased from Sigma Aldrich. The standard and internal standard are first dissolved in acetonitrile, and then stored at -80°C. When the actual sample detection is required, the internal standard deuterated cholic acid is diluted to 76nmol / L with acetonitrile, and the internal standard deuteoxycholic acid is diluted to 76nmol / L. The concentration is 104nmol / L, and the dilution concentration of internal standard deuterated ursodeoxycholic acid is 126nmol / L, which is used as the working solution.

[0041] (1) Standard product preparation: Take 0.1mL of 9 different concentrations of b...

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Abstract

The invention discloses a method for detecting five free bile acids in serum through high performance liquid chromatography-tandem mass spectrum. The five free bile acids comprises a cholic acid, a lithocholic acid, a deoxycholic acid, a ursodeoxycholic acid and a chenodesoxycholic acid; the method comprises the following steps of preparing a standard substance, performing high performance liquid chromatography separation, performing tandem mass spectrum detection, and detecting the free bile acids in the serum. The method can be used to simultaneously perform determination and accurately quantify the content of the free bile acids in human serum, the detection time is short, the flux is high, the detection sensitivity is high, and the specificity is good.

Description

technical field [0001] The invention relates to the field of bile acid detection, in particular to a method for detecting five kinds of free bile acids by high performance liquid chromatography tandem mass spectrometry. Background technique [0002] Bile acid, the main organic component of bile, is a general term for a class of 24-carbon cholic acid hydroxyl derivatives. Bile acids can be divided into two types according to their structure: free bile acid and conjugated bile acid. Free bile acids in the human body are divided into primary and secondary bile acids according to their production sites and structures. Primary bile acids are converted from intrahepatic cholesterol, and through a series of reactions, finally generate cholic acid (CA) and chenodeoxycholic aic (CDCA). After the primary bile acid is synthesized in the liver cells, a series of transport proteins in the capillary membrane of the liver cells enter the capillary bile duct in the form of bile salts and ...

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Application Information

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IPC IPC(8): G01N30/88G01N30/06
CPCG01N30/06G01N30/88G01N2030/8822
Inventor 吴超超
Owner HANGZHOU BAICHEN MEDICAL LAB CO LTD
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