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Total bile acid quantitative determination method and determination reagent kit

A technology for quantitative determination of total bile acids, which can be used in biological testing, color/spectral property measurement, material inspection products, etc.

Inactive Publication Date: 2012-07-04
宁波天康生物科技有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

After the search of relevant patents, no patent was found on the reagent (box) for the determination of TBA by the fifth-generation enzyme cycle method

Method used

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  • Total bile acid quantitative determination method and determination reagent kit
  • Total bile acid quantitative determination method and determination reagent kit
  • Total bile acid quantitative determination method and determination reagent kit

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0040] One, prepare following reagent 1 and reagent 2 of the present invention according to following composition and ratio:

[0041] Reagent 1:

[0042]

[0043] Reagent 2:

[0044]

[0045] 2. The determination uses the HITACHI7100 automatic biochemical analyzer, and the analysis parameters are set as follows: analysis method: rate method; measurement point: 19, 28; detection wavelength 405nm (main) / 660nm (secondary); sample volume: 3.0μl; reagent R1 : 210μl; Reagent R2: 70μl; Reaction direction: rising; Calibration mode: linear mode, 2-point calibration; Unit: μmol / L. The instrument automatically performs the following operation process: first add 3 μl of the serum sample to be tested into the cuvette, then add 1210 μl of the reagent, mix well, and keep it in a constant temperature environment at 37°C for 5 minutes at regular intervals, then add reagent 2 to start the enzyme reaction, Continue to keep warm, and after a short delay time, start to continuously monitor...

Embodiment 2

[0054] Using the reagents listed in this Example 1, the TBA content of 20 serum samples was measured according to the method and conditions described in Example 1, and each serum sample was simultaneously tested with a commercially available TBA assay kit (Yantai Ausbang Biology) The product of Engineering Co., Ltd., batch number 0911.2) carries out comparative timing determination, and the results are listed in Table 2 below:

[0055] Table 2 The assay result of reagent of the present invention and contrast reagent to TBA

[0056]

[0057]

[0058] After statistical analysis of the experimental data, we get:

[0059] Related equation: Y=1.03x+0.2067(n=20)

[0060] R 2 =0.9985 or R=0.9994 (analyzed by statistical analysis software)

[0061] The accuracy of the method of the present invention is measured by a correlation experiment with the control method. Gained correlation equation Y=1.03x+0.2067 from the statistical result of experimental data can find out that slo...

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Abstract

The invention provides a reagent for quantitatively determining content of total bile acid in a human serum sample through an enzymological method. The reagent is suitable for an automatic biochemical analyzer to automatically and quantitatively determine the total bile acid. The reagent consists of a solution reagent 1 and a reagent 2 which are separately placed, wherein the reagent 1 contains oxidized thio-nicotinamide-adenine dinucleotide (Thio-NAD), buffer and stabilizer; and the reagent 2 contains reduced coenzyme I (NADH), hydroxysteroiddehydrogenase (HSD), stabilizer and buffer. The invention additionally provides a reagent kit with the reagent 1 and the reagent 2, and a method for determining the content of total bile acid in the serum. The reagent, the reagent kit and the method provided by the invention have the advantages of high sensitivity, low cost, simplicity, convenience and rapidness in operation and easiness in popularization.

Description

technical field [0001] The invention relates to a quantitative determination method, reagent and kit of total bile acid (TBA). Background technique [0002] Bile acids are 24-carbon steroid compounds produced in the liver from cholesterol. There are five main forms of bile acids in body fluids: cholic acid, chenodeoxycholic acid, deoxycholic acid, lithocholic acid, and ursodeoxycholic acid. These bile acids are collectively referred to as total bile acids (TBA). Serum bile acid levels are an important indicator of liver parenchyma damage. The postprandial TBA assay is more sensitive than all other liver function tests for detecting mild liver disease. Sensitivity for detecting liver cell damage caused by alcohol or industrial chemicals is superior to other liver function tests. [0003] Most bile acid determination methods require a separation and extraction step before determination, which is due to factors such as bile acid insolubility, polymorphism, and existence in b...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/96G01N21/31
Inventor 李清华宋高峰
Owner 宁波天康生物科技有限公司
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