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Determination method of serum total bile acid

A technology of serum total bile acid and determination method, which is applied in the direction of measuring device, material analysis by electromagnetic means, instrument, etc., can solve the problems of further improvement of sensitivity, high cost of total bile acid, unsuitable for clinical detection, etc. Beneficial for clinical promotion, simple sample pretreatment, and fewer types of reagents

Inactive Publication Date: 2010-09-08
CHONGQING MEDICAL UNIVERSITY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

Derivatization and hydrolysis are required for the determination of total bile acids by gas or liquid chromatography, which is cumbersome and not suitable for clinical testing
At present, most of the CE detection of bile acids uses ultraviolet detection, and the detection sensitivity is low, which limits its application.
However, the enzymatic cycle amplification method used clinically to determine total bile acids is expensive, and the sensitivity needs to be further improved.

Method used

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  • Determination method of serum total bile acid
  • Determination method of serum total bile acid
  • Determination method of serum total bile acid

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0039]This embodiment is to investigate the influence of the pH of the buffer solution in the detection cell on the ECL signal. Experimental conditions: put the electrochemiluminescence detection cell into the dark box of the luminescence detector, so that the light transmission window is facing the photomultiplier tube (such as figure 1 ), the high voltage of the photomultiplier tube is -700V. The electrochemiluminescence detection cell adopts a three-electrode system: the working electrode is a platinum disc electrode with a diameter of 500 μm, the auxiliary electrode is a platinum electrode with a diameter of 1 mm, and the reference electrode is an Ag / AgCl electrode with a diameter of 300 μm (filled with saturated KCl solution), the detection voltage is 1.17V. The inner diameter of the open quartz capillary is 25 μm, the outer diameter is 365 μm, the total length is 50 cm, and the effective length is 45 cm. The running buffer contains 5nmol / L NAD + 5mmol / L PBS pH 7.0. P...

Embodiment 2

[0041] This embodiment is to investigate the influence of the PBS concentration in the detection cell on the ECL signal. Based on the above conditions, the pH value of the buffer solution was set at 8.0, and the concentration of the buffer solution was changed within the range of 25-100mmol / L to investigate its influence on the ECL intensity. The results are as follows image 3 . When the phosphate concentration does not exceed 75mmol / L, the ECL intensity of the system increases with the increase of the buffer solution concentration, and the signal stability is good; ratio becomes smaller. Therefore, the concentration of the phosphate buffer solution in the detection cell was chosen to be 70-75mmol / L in the experiment.

Embodiment 3

[0043] This embodiment is to investigate Ru(bpy) in the detection pool 3 2+ Effect of concentration on ECL intensity. Based on the above conditions, change Ru(bpy) in the range of 1~20mmol / L 3 2+ Concentration, examine Ru(bpy) 3 2+ The influence of concentration on the sensitivity of the present invention, such as Figure 4 shown. ECL intensity increases with Ru(bpy) 3 2+ When the concentration increases, the luminescent signal intensity decreases when the concentration exceeds 5mmol / L. Ru(bpy) in detection pool 3 2+ Set to 3 ~ 5mmol / L.

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Abstract

The invention relates to a determination method of serum total bile acid, which is characterized in that: bile acid is collected by a capillary electrophoresis serum sample, bile acid in a column-end detector cell is dehydrogenated through the catalysis of steroid dehydrogenase (3 alpha-HSD) with the coexistence of oxidized coenzyme I (NAD) +, and NAD + is reduced to nicotinamide adenine dinucleotide I (NADH); and the generated NADH is oxidized to NAD + by tris (bipyridine) ruthenium (Ru (bpy) 32 +) under the action of constant potential, simultaneously photon is released and recorded, thereby determining total bile acid in a conventional way. The method of the invention significantly reduces the amount of expensive enzyme reagent, chemical reagent and biological sample, decreasing the waste liquid, remarkably improving the sensitivity, and realizing the rapid determination of discontinuous batch of clinical samples.

Description

technical field [0001] The invention relates to a method for measuring total bile acids in serum, in particular to an analysis method for measuring total bile acids in serum by electrochemiluminescence, which belongs to the field of clinical biochemical analysis. Background technique [0002] Bile acid (BA) is the main metabolite of endogenous cholesterol in the liver. It is a general term for a class of cholanic acid that exists in bile. It exists in the form of sodium salt or potassium salt. The serum level is the only serological indicator that can simultaneously reflect the three aspects of liver secretion, liver synthesis and metabolism, and liver cell damage. In liver disease, liver cell damage can cause bile acid synthesis and excretion disorders, cholesterol 7α-hydroxylase and 12α-hydroxylase activities in liver cells are reduced, and bile acid synthesis is significantly reduced. The hepatocyte uptake of bile acid dysfunction slows down the clearance rate of bile ac...

Claims

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Application Information

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IPC IPC(8): G01N27/447
Inventor 丁敏邓文平王箭张晓清
Owner CHONGQING MEDICAL UNIVERSITY
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