Unlock instant, AI-driven research and patent intelligence for your innovation.

Serum total bile acid detection kit and its application method

A serum total bile acid and detection kit technology, applied in the field of medical detection, can solve the problems of the stability of biological reagents, increase the detection time, and require special personnel to operate, so as to avoid cross-contamination in experiments, avoid grinding and polishing, and be mobile. strong effect

Inactive Publication Date: 2015-12-30
CHONGQING MEDICAL UNIVERSITY
View PDF4 Cites 0 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] (1) Mass spectrometry can accurately determine the molecular weight of bile acids and classify bile acids, but the structure of bile acids is relatively complex, and tedious analysis of the results is required
The detection instrument is expensive, the operation is complicated and requires special personnel to operate, and the sample needs to be purified to avoid clogging the instrument
And there are problems such as memory effect and pollution caused by ion source
[0005] (2) Chromatography can separate and quantitatively analyze serum bile acids well, but the sample needs to be derivatized and hydrolyzed before the determination of serum bile acids, which complicates the whole detection process and increases the detection time, which is not suitable for wide clinical application
[0006] (3) The radioimmunoassay method is simple to operate, but the stability of its biological reagents will be affected by many factors, and there is radioactive pollution, which will cause unpredictable harm to the environment and staff
[0007] (4) Enzyme cycling method has been used clinically, but the reagents are expensive, which may easily cause contamination of testing instruments and affect the measurement results, and requires special testing instruments, which are expensive and difficult to move
At present, there are no relevant research reports on the kit and its application method for the detection of serum total bile acids by electrochemical analysis method at home and abroad

Method used

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
View more

Image

Smart Image Click on the blue labels to locate them in the text.
Viewing Examples
Smart Image
  • Serum total bile acid detection kit and its application method
  • Serum total bile acid detection kit and its application method
  • Serum total bile acid detection kit and its application method

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0049] Example 1: Preparation of Serum Total Bile Acid Detection Kit

[0050] 1) Purchase analytically pure methanol and place it in a container;

[0051] 2) Preparation of phosphate buffer: prepare 0.2mol / L disodium hydrogen phosphate solution, then add saturated sodium hydroxide solution to adjust to pH = 12.2, and place in a container;

[0052] 3) Preparation of 40U / mL 3α-HSD: Prepare the purchased 3α-HSD into a solution with an activity of 40U / mL and place it in a container;

[0053] 4) Prepare 100mmol / L NAD + : NAD to be purchased + Prepare a solution with a concentration of 100mmol / L and place it in a container;

[0054] 5) Prepare the screen printing electrode of the present invention:

[0055] The screen-printed electrode of the present invention prints carbon paste, silver paste and insulating paste sequentially on a polyethylene terephthalate (PET) substrate 5 . Specifically include the following steps:

[0056] ① Clean the PET substrate 5, print carbon paste o...

Embodiment 2

[0061] Example 2: Use of Serum Total Bile Acid Detection Kit

[0062] 1) thawing the serum total bile acid detection kit at room temperature;

[0063] 2) Take 100 μL serum sample in EP tube (centrifuge tube), add 300 μL methanol dropwise under vortex condition to remove protein in serum;

[0064] 3) Centrifuge the protein-removed serum sample at 12,000×g for 10 min, take the supernatant and dry it with nitrogen at 60°C, and redissolve it with 25 μL of 0.2 mol / L, pH=12.2 phosphate buffer;

[0065] 4) Take 10 μL of the solution obtained in step 3), 1 μL of 40 U / mL 3α-HSD and 10 μL of 100 mmol / L NAD + Vortex and mix well, the activity of 3α-HSD in the mixed solution is 2U / mL, NAD + The concentration of the bile acid enzyme was 50 mmol / L, and the enzymatic reaction was carried out at room temperature for 10 minutes to obtain the bile acid enzyme catalyzed reaction mixture;

[0066] 5) Connect the screen printing electrode to CHI852C electrochemical workstation, and set its para...

Embodiment 3

[0068] This example is to investigate the electrochemical response of the screen-printed electrode to NADH, an enzyme-catalyzed reaction product of bile acids. Measure the blank serum (the human serum sample whose bile acid concentration is less than 0.5 μmol / L in the middle of the present invention) and the spiked serum (the serum sample added with the bile acid standard in the middle of the present invention), the operation steps are the same as in Example 2, and the results are shown in figure 2 . The solid line in the figure is the differential pulse voltammetry curve of the spiked serum, and the dotted line is the differential pulse voltammetry curve of the blank serum, indicating that the electrochemical signal generated is the electrochemical signal of NADH, the enzymatic reaction product of bile acid.

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
Login to View More

PUM

No PUM Login to View More

Abstract

The invention discloses a serum total bile acid detection kit and a using method thereof. The kit comprises a serum sample pretreatment solution, a phosphate buffer, a 3 alpha-hydroxysteroid dehydrogenase (3 alpha-HSD), beta-nicotinamide adenine dinucleotide trihydrate I (NAD<+>) and a screen-printed electrode. According to the serum total bile acid detection kit, an electrochemical detection is performed by adopting the screen-printed electrode, and the serum total bile acid can be quickly and accurately detected indirectly through an electrochemical analysis method. The serum total bile acid detection kit is simple in reagent, low in price, convenient to use and high in cost performance.

Description

technical field [0001] The invention belongs to medical detection technology, and mainly relates to a serum total bile acid detection kit and a use method thereof. Background technique [0002] Bile acids are the metabolites of cholesterol. They are mainly synthesized in the liver through the classical pathway of cholesterol 7α-dehydroxylase, secreted into the hepatic bile duct and stored in the gallbladder. After being stimulated by food, they are discharged into the intestinal tract and enter the hepatic portal vein through the enterohepatic circulation. Reabsorbed by hepatocytes. Bile acids can promote the digestion and absorption of lipids and increase the solubility of cholesterol to prevent gallstone formation. The increase of bile acid is mainly related to liver and gallbladder diseases, and it is also increased to varying degrees in other diseases such as intrahepatic cholestasis of pregnancy (ICP). At present, it is considered that the serum total bile acid level ...

Claims

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
Login to View More

Application Information

Patent Timeline
no application Login to View More
Patent Type & Authority Patents(China)
IPC IPC(8): G01N27/48
Inventor 丁敏罗娟张晓清朱明松崔悦左明
Owner CHONGQING MEDICAL UNIVERSITY
Features
  • R&D
  • Intellectual Property
  • Life Sciences
  • Materials
  • Tech Scout
Why Patsnap Eureka
  • Unparalleled Data Quality
  • Higher Quality Content
  • 60% Fewer Hallucinations
Social media
Patsnap Eureka Blog
Learn More

Browse by: Latest US Patents, China's latest patents, Technical Efficacy Thesaurus, Application Domain, Technology Topic, Popular Technical Reports.

© 2025 PatSnap. All rights reserved.Legal|Privacy policy|Modern Slavery Act Transparency Statement|Sitemap|About US| Contact US: help@patsnap.com