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Method and kit for simultaneously detecting various bile acids in blood sample

A technology for blood samples and bile acids, which is applied to measurement devices, instruments, scientific instruments, etc., can solve the problems of lack of reference range for healthy people, low method throughput, etc., and achieves the effect of saving time, simple and fast operation, and reliable results.

Pending Publication Date: 2021-06-22
广东南芯医疗科技有限公司 +1
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  • Claims
  • Application Information

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Problems solved by technology

However, there are relatively few studies on the detection of bile acid subtypes in human serum, and the throughput of the method is low, and there is also a lack of reference ranges for healthy people.

Method used

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  • Method and kit for simultaneously detecting various bile acids in blood sample
  • Method and kit for simultaneously detecting various bile acids in blood sample
  • Method and kit for simultaneously detecting various bile acids in blood sample

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Embodiment 1

[0054] A method for simultaneously detecting multiple bile acids in a blood sample, the steps are as follows:

[0055] (1) Prepare the sample to be tested

[0056] After taking the sample and vortexing for 30s, take 400μl of the suspension sample in a 1.5ml Ep tube, centrifuge at 13000rpm at 4°C for 15min; take 100μL of the supernatant, put it in a 1.5mL Ep tube, add 100μL of methanol to dilute; add 1μL of N-1- Naphthylphthalic acid internal standard solution (0.033mM) was mixed and then filtered with a 0.22μm microporous membrane, and all samples were transferred to a 96-well plate, covered with aluminum foil, and used for detection;

[0057] (2) Ultra-high performance liquid chromatography-mass spectrometry detection

[0058] The sample to be tested enters the chromatographic column for separation through the gradient elution mode, and the reference conditions of liquid chromatography are:

[0059] Chromatographic column: Waters ACQUITYUPLC HSS T3 (1.8μm, 2.1×100mm)

[00...

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Abstract

The invention discloses a method and a kit for simultaneously detecting various bile acids in a blood sample, and belongs to the technical field of bile acid detection. The invention discloses a method for simultaneously detecting various bile acids in a blood sample, which comprises the following steps: pretreating the sample, performing chromatographic separation and mass spectrometric detection, respectively selecting a pair of qualitative ions and a pair of quantitative ions for each bile acid, and determining the content of various bile acids in the blood sample by taking the relative retention time of each bile acid and the qualitative ion pair as a qualitative basis; and making a standard curve by using a standard product for quantification. Meanwhile, the method provided by the invention is accurate and effective by applying three levels of quality control product inspection methods, and detection result distortion is avoided. According to the method, the aim of simultaneously detecting fifteen bile acids in a serum sample by applying the LC-MS technology is achieved for the first time, the influence of interferents is reduced, the method is simple, convenient and rapid to operate, the bile acid level in a human body is effectively monitored, and the method has guiding significance on reasonable and safe supplement of the bile acids and is easy to clinically popularize and popularize.

Description

technical field [0001] The invention relates to the technical field of bile acid detection, in particular to a method and a kit for simultaneously detecting multiple bile acids in blood samples. Background technique [0002] Bile acids are produced by the metabolism of cholesterol. According to the synthesis route, they can be divided into primary bile acids directly synthesized from cholesterol and secondary bile acids produced by metabolism. Bile acid is an important component of bile and an essential substance for human fat metabolism. Bile acids can not only regulate glucose and fat metabolism, but also interact with intestinal flora, which has important clinical significance. Existing detection methods can only simply quantitatively measure the level of total bile acids, but cannot quantitatively detect the levels of each subtype of bile acids in the body. [0003] In the prior art, commonly used bile acid detection methods mainly include gas chromatography, liquid ch...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N30/88G01N30/06G01N30/14
CPCG01N30/88G01N30/06G01N30/14G01N2030/8822G01N2030/027G01N2030/146
Inventor 张召刁嘉茵郑康帝陈涛
Owner 广东南芯医疗科技有限公司
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