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Method and kit for detecting content of bilirubin

A detection method and technology for bilirubin, which are applied in the field of analysis and detection, can solve problems such as direct quantitative determination of bilirubin, etc., and achieve the effects of reliable detection results, low detection limit and convenient sampling.

Pending Publication Date: 2022-06-21
HUMAN METABOLOMICS INST INC
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] Quantitative determination of indirect bilirubin and direct bilirubin is provided in the above prior art, but there is no quantitative determination of each component of direct bilirubin

Method used

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  • Method and kit for detecting content of bilirubin
  • Method and kit for detecting content of bilirubin
  • Method and kit for detecting content of bilirubin

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0068] Example 1 Establishment and optimization of liquid chromatography-tandem mass spectrometry method

[0069] In this example, the establishment of a method for qualitative and quantitative detection of bilirubin components in biological samples based on liquid chromatography-tandem mass spectrometry is described for illustrative purposes, including selecting liquid phase conditions, setting mass spectrometry conditions, and preparing standard solutions , optimize the analysis method, draw the standard curve.

[0070] Bilirubin samples contain a variety of components. The inventor found through research that when the pH value of the mobile phase A of liquid chromatography is 3-4.5, the main components in the bilirubin sample are free bilirubin, biliverdin, bilirubin monoglucuronide and bilirubin. It is suitable for a wide range of chromatographic columns, especially C8 and C18 silica gel columns, and has good applicability to column temperature and injection volume.

[0...

Embodiment 2

[0095] Example 2 Qualitative and quantitative determination of bilirubin in blood samples

[0096] Sampling: Take serum, plasma and whole blood samples of 3 test subjects, and each sample is divided into 3 parts, such as serum 01, serum 02, and serum 03, a total of 9 biological samples.

[0097] Biological sample processing:

[0098] Serum 01: Take 20 microliters of samples, add 10 microliters of taurine bilirubin as the internal standard, and then add 80 microliters of methanol:acetonitrile (volume ratio 1:1) solution containing 1mg / ml BHT and 200mmol / L ascorbic acid , shake at 1450rpm for 20 minutes in the dark at 10°C. Centrifuge at 18,000g for 20 minutes at 4°C. 60 microliters of supernatant was taken into a 96-well plate as a detection sample for high-performance liquid chromatography tandem mass spectrometry detection.

[0099] Serum 02: Take 20 microliters of samples, add 10 microliters of taurine bilirubin as internal standard, and then add 80 microliters of methano...

Embodiment 3

[0110] Example 3 Qualitative and quantitative determination of bilirubin components in dried blood spots

[0111] Dried blood spot sample preparation

[0112] Dried blood spot samples were prepared by collecting fingertip blood within 4 days of birth. Proceed as follows:

[0113] a) Pretreatment of filter paper: Soak the 903 filter paper in an ethanol solution containing 1 mg / mL BHT and 200 mmol / L ascorbic acid for 2 minutes, take it out and drain, and then naturally dry the filter paper for later use.

[0114] b) Preparation: Take a suitable test tube and add an appropriate amount of diluent. Connect the micropipette to the latex pipette, check whether the connection is leaking, or take a disposable micropipette (siphon principle), blood collection needle, 75% ethanol or iodophor, cotton swab, etc.

[0115] c) Massage: Gently massage the central part of the left ring finger with more muscles on the ulnar side of the finger, so that the local tissue is naturally congested. ...

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Abstract

The invention discloses a bilirubin content detection method. The detection method is a liquid chromatography-tandem mass spectrometry detection method, and the content of bilirubin comprises the content of any one or more of free bilirubin, biliverdin, bilirubin monoglucuronate and bilirubin diglucuronate or the sum of the content of the free bilirubin, the biliverdin, the bilirubin monoglucuronate and the bilirubin diglucuronate. According to the liquid chromatography-tandem mass spectrometry detection method, liquid chromatography is selected from high performance liquid chromatography, ultra-high performance liquid chromatography and nano-liter liquid chromatography. The content detection method disclosed by the invention can be used for qualitatively and quantitatively detecting one or more of the contents of free bilirubin, biliverdin, bilirubin monoglucuronate and bilirubin diglucuronate simultaneously or respectively; compared with a traditional biochemical method, the content detection method disclosed by the invention has the characteristics of rapidness in operation, high accuracy and low detection limit.

Description

technical field [0001] The invention relates to the technical field of analysis and detection, in particular to a qualitative and quantitative detection and analysis method of bilirubin, which can be applied to the qualitative and quantitative detection of bilirubin in a test sample. Background technique [0002] Bilirubin is formed by deferrication of heme released after red blood cells are destroyed. 80% of bilirubin comes from the spleen and liver, and the rest comes from the bone marrow. Bilirubin exists in three forms, indirect bilirubin or unconjugated bilirubin, conjugated bilirubin, and delta bilirubin. Among them, conjugated bilirubin and delta bilirubin can react directly with the diazo reagent, which is called direct bilirubin. Direct bilirubin, also known as conjugated bilirubin, is formed by the combination of glucuronic acid and glucuronic acid in the liver after indirect bilirubin enters the liver. Bilirubin is an important basis for clinical judgment of jau...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N30/88
CPCG01N30/88G01N2030/8822
Inventor 贾伟谢国祥周科军
Owner HUMAN METABOLOMICS INST INC
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