Ultra-high performance liquid chromatography-tandem mass spectrometry detection method

A kind of ultra-high performance liquid phase, detection method technology, applied in the field of biological analysis and detection

Active Publication Date: 2017-08-11
苏长青
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Problems solved by technology

[0005] However, there is no rapid, accurate, specific and sensitive method for the simultaneous quantification of bile acids, lysolecithin and unsaturated fatty acids in body fluids

Method used

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Embodiment 1

[0032] 1. Materials and Methods

[0033] 1.1 Reagents

[0034] Milli-Q pure water (Millipore, USA), chromatographically pure organic reagents (Aladdin, China) were used for protein precipitation, and mass spectrometrically pure reagents (Merck, Germany) were used for liquid chromatography-mass spectrometry analysis experiments. Eight bile acid standards, four lysolecithin standards, three polyunsaturated fatty acid standards and two internal standards (see Table 1) were purchased from Sigma-Aldrich.

[0035] Table 1. Details of target metabolites and internal standards

[0036]

[0037]

[0038] 1.2 Collection of plasma, serum and urine samples

[0039] The blood samples of four patients with liver cancer or cholangiocarcinoma and four healthy people were collected with anticoagulant tubes and procoagulant tubes respectively, and the whole blood collected in the anticoagulant tubes was centrifuged at low temperature (4°C, 1000g) for 10 minutes, and the supernatant plas...

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Abstract

The invention relates to an ultra-high performance liquid chromatography-tandem mass spectrometry detection method. On the basis of an ultra-high performance liquid chromatography-tandem mass spectrometry detection platform, firstly, mass spectrometry detection parameters and chromatography separation conditions are optimized one by one by use of standard substances of target metabolites; an extraction method of 15 key target metabolites (including 4 lysolecithin, 3 unsaturated fatty acids and 8 bile acids) in blood plasma is further explored with an optimized chromatography-mass spectrometry method; LOD (limit of detection), LOQ (limit of quantitation), degree of precision, accuracy, the linear range and the like of the metabolites, detected simultaneously, in the blood plasma and urine are studied with an optimized analysis method. Results prove that the detection method can rapidly, accurately, specifically and sensitively quantify the content of the 15 key target metabolites in a body fluid, and a foundation is laid for screening of non-intrusive clinical diagnostic markers of liver and gall diseases.

Description

technical field [0001] The invention relates to the technical field of biological analysis and detection, in particular to a detection method of ultra-high performance liquid chromatography tandem mass spectrometry. Background technique [0002] The liver is the most important metabolic organ for endogenous and exogenous small molecule metabolites in the body. A large number of studies have shown that with the progression of liver disease (normal liver-hepatitis-cirrhosis-hepatic cancer), the metabolic homeostasis of bile acids, lysolecithin and unsaturated fatty acids will be disrupted, and the three levels corresponding to different degrees of liver damage The metabolic characteristics of metabolites are also different. Therefore, if the content of these three metabolites in body fluids can be quantitatively analyzed, it is of great significance for the screening of small molecule diagnostic markers for hepatobiliary diseases. Individual quantification methods for each of...

Claims

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Application Information

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IPC IPC(8): G01N30/02G01N30/72
CPCG01N30/02G01N30/7266
Inventor 苏长青彭章晓林雪晶钱海华
Owner 苏长青
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