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Kit for detecting total bile acid in urine by electrochemical method and application thereof

A total bile acid, electrochemical technology, applied in the field of electrochemical detection, can solve the problems of large volume and expensive special equipment, and achieve the effects of high sensitivity, accurate quantification, and low production cost

Pending Publication Date: 2022-04-15
重庆医科大学国际体外诊断研究院
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

This method is widely used clinically, but its detection sensitivity is at the μmol / L level, which needs to be improved, and the special instruments used for detection are expensive and bulky

Method used

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  • Kit for detecting total bile acid in urine by electrochemical method and application thereof
  • Kit for detecting total bile acid in urine by electrochemical method and application thereof
  • Kit for detecting total bile acid in urine by electrochemical method and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0025] Embodiment 1: the preparation of screen printing carbon electrode of the present invention

[0026] The screen printing carbon electrode of the present invention is to sequentially print carbon paste, Ag / AgCl paste and insulating paste on a PET substrate. Specifically include the following steps:

[0027] 1) Clean the PET substrate, print carbon paste on the PET substrate after drying, make carbon working electrode and carbon paste auxiliary electrode, and dry at room temperature;

[0028] 2) printing silver paste containing silver chloride on the above-mentioned PET substrate to make Ag / AgCl reference electrode, and drying at normal temperature;

[0029] 3) Avoid the circular working area, and print insulating paste on the above-mentioned PET substrate to cover the wire;

[0030] 4) The above-mentioned working electrode, auxiliary electrode and reference electrode form a circular working area, and each electrode is connected to the interface through a wire under the ...

Embodiment 2

[0032] Embodiment 2: The specific steps of using the kit for detecting total bile acids in urine by electrochemical method according to the present invention:

[0033] 1) Pretreatment of urine samples: Take 10 μL of urine and 990 μL of ultrapure water and add 1.5 mL of E P Vortex and mix well in the tube to obtain a diluted urine sample; then take 10 μL and 990 μL of ultrapure water and add 1.5 mL of diluted urine sample P Vortex and mix well in the tube to obtain the second diluted urine sample;

[0034] 2) Take 8μL 100mmol / L NAD respectively + , 12μL 10kU / L 3α-HSD, 7μL 100mmol / L Ru(bpy) 3 2+ , 23μL ultrapure water and 50μL PBS solution with a concentration of 100.0mmol / L, the pH value is 8.5, add 1.5mL E P Vortex and mix well in the tube to obtain the detection mixture;

[0035]3) Take 50 μL of the mixed solution obtained in step 2 and add it dropwise to the working area of ​​SPCE. The SPCE is connected to the electrochemical workstation, and the current-time (I-t) curv...

Embodiment 3

[0037] This embodiment investigates the correlation between the oxidation peak current of the electrochemical detection kit and the urine bile acid concentration, and the results are as follows figure 2 shown. Add a series of concentration (1.0-100.0μmol / L) bile acid standard solution to the urine matrix, after 10 4 After two-fold dilution, the assay was carried out under the optimal detection conditions. In the concentration range of 1.0-100.0 μmol / L, the oxidation peak current has a good linear relationship with the logarithm of the bile acid concentration, and the regression equation is: I=2.3065lgc+8.182, R 2 = 0.9922. LOD was 0.2 μmol / L (S / N=3).

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Abstract

The invention discloses a kit for detecting total bile acid in urine by an electrochemical method and application of the kit. The kit for detecting the total bile acid in the urine by the electrochemical method comprises the following reagents: a phosphate buffer solution, 3 alpha-hydroxysteroid dehydrogenase (3 alpha-HSD), oxidized coenzyme I (NAD < + >), terpyridyl ruthenium (Ru (bpy) 3 < 2 + >) and a screen-printed electrode. According to the kit for detecting the total bile acid in the urine by the electrochemical method, a screen-printed electrode is combined with an I-t curve, and rapid quantitative detection of the total bile acid in the urine is realized according to a linear relationship between an electrochemical response signal and the logarithm of the concentration of an object to be detected. The electrochemical detection kit is simple, convenient and rapid, has good repeatability, stability and accuracy, can realize rapid quantitative detection of the total bile acid content in urine, and provides a new choice for non-invasive real-time monitoring of liver and gall diseases.

Description

technical field [0001] The invention belongs to the field of electrochemical detection, and mainly relates to a kit for detecting total bile acids in urine by an electrochemical method and an application thereof. Background technique [0002] Bile acids are a class of 24-carbon cholic acids that are synthesized in the liver from cholesterol. Bile acids have important physiological functions. They can not only promote the digestion and absorption of fat, but also act as signal molecules to activate various receptors, mediate various signal regulation pathways, and play a role in the body's glucose, fat, self-metabolism and energy metabolism. important role. In liver disease, damage to liver cells can cause bile acid synthesis and excretion disorders, and the synthesis of bile acids is significantly reduced. Dysfunction of bile acid uptake by liver cells slows down the clearance of bile acids from blood, resulting in increased bile acid content in blood and urine. Serum bil...

Claims

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Application Information

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IPC IPC(8): G01N27/30
CPCG01N27/308
Inventor 张晓清李彦茹丁敏吕文静朱阳宋紫苹
Owner 重庆医科大学国际体外诊断研究院
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