Looking for breakthrough ideas for innovation challenges? Try Patsnap Eureka!

Quantitative determination method and kit for malondialdehyde content in seminal plasma

A technology for quantitative determination of malondialdehyde, applied in the field of in vitro detection of sperm biochemical indicators, can solve the problems of large error in test results, cumbersome operation, high test results, etc., and achieve the effect of increasing absorbance value, increasing sensitivity and eliminating influence

Pending Publication Date: 2021-01-22
珠海高瑞特医疗科技有限公司
View PDF4 Cites 0 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0003] In the prior art, the invention patent with the application number CN03818036.7 discloses a kit for the determination of MDA in urine. Although it can detect the presence of MDA in urine, it is limited to qualitative measurement and cannot be quantitatively measured; while the application number is CN201910985706 The invention patent of .0 discloses a reagent strip for detecting MDA in urine. The detection result is analyzed by a chemical analyzer to obtain the content of MDA in urine, but this product cannot eliminate the impact of soluble sugar and unsaturated fatty acid in the specimen. The influence of MDA determination, the error of the test result is large, and the clinical application value is low
[0004] The invention patent with the application number CN108572224A discloses a method for measuring the content of malondialdehyde in biological tissues. The method adopts the use of acridone acetylhydrazide as a fluorescent labeling reagent under the catalysis of trichloroacetic acid solution to extract the biological tissue and The malondialdehyde in the malondialdehyde standard solution is derivatized, and the high-performance liquid chromatography with a fluorescence detector is used for quantitative detection and analysis. Although the detection accuracy of this method is high, the operation is cumbersome and requires a high-performance liquid chromatography detection, not suitable for promotion
[0005] The invention with the application number CN201910242240.5 discloses a method for determining the content of malondialdehyde in catalpa leaves. The TBA method is used to detect MDA, but this patent is applicable to the determination of malondialdehyde content in plant leaf specimens such as catalpa leaves. , the sample processing method is quite different from that of seminal plasma, so it is not suitable for the determination of MDA content in seminal plasma
In addition, because human seminal plasma samples contain a certain amount of fructose, which will interfere with the TBA reaction, soluble sugars such as fructose in seminal plasma samples can also react with TBA reagents, and the reaction product has an absorption peak at 450nm, and at 532nm Absorbance will also be generated, which will make the MDA measurement results higher. This patented technology does not provide a method to eliminate the influence of soluble sugars such as fructose on the detection results, and is not suitable for the determination of malondialdehyde content in human seminal plasma.

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
  • Quantitative determination method and kit for malondialdehyde content in seminal plasma
  • Quantitative determination method and kit for malondialdehyde content in seminal plasma
  • Quantitative determination method and kit for malondialdehyde content in seminal plasma

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0027] Reagent composition: 10umol / L malondialdehyde solution (standard product), 2mol / L trichloroacetic acid (specimen treatment solution), TBA reagent.

[0028] Reagent preparation:

[0029] Preparation of 0.02mol / L hydrochloric acid: Measure 10mL of 0.24mol / L hydrochloric acid and dilute with water to 120mL.

[0030] Preparation of malondialdehyde standard (10umol / L) (molecular weight: 164.2): Accurately weigh 16.42mg of malondialdehyde dimethyl acetal (TMP), add 0.02mol / L hydrochloric acid 1000ml, stir at room temperature for 60min, after TMP is fully dissolved , aliquoted into 0.5ml / bottle, stored at 2-8°C.

[0031] Preparation of specimen treatment solution: Accurately weigh 16.34g of trichloroacetic acid, dissolve in 50ml of deionized water, and store at 2-8°C.

[0032] Preparation of 1mol / L sodium hydroxide solution: Accurately weigh 4g of sodium hydroxide, dissolve in 100ml of deionized water, and store at 2-8°C.

[0033] Preparation of 100mg / L ferric sulfate solut...

Embodiment 2

[0074] A quantitative detection kit for malondialdehyde content in seminal plasma, comprising, malondialdehyde standard product, specimen treatment solution and TBA reagent; wherein, the concentration of the malondialdehyde standard product is 10umol / L, and the specimen treatment solution is 2mol / L trichloroacetic acid, the TBA reagent includes 0.8% thiobarbituric acid solution and 0.1mg / L ferric sulfate solution. The kit includes a centrifuge tube with a specification of 0.5-5.0ml, and the malondialdehyde standard, specimen processing solution and TBA reagent are separately packed in bottles.

[0075] In the above example, the preparation steps of malondialdehyde standard (10umol / L): Accurately weigh 16.42mg of malondialdehyde dimethyl acetal (TMP), add 1000ml of 0.02mol / L hydrochloric acid, fully stir at room temperature for 60min, and wait until TMP is fully dissolved Finally, aliquot into 0.5ml / bottle and store at 2-8°C.

[0076] Preparation steps of specimen treatment so...

Embodiment 3

[0080] A quantitative detection kit for malondialdehyde content in seminal plasma, comprising, malondialdehyde standard product, specimen treatment solution and TBA reagent; wherein, the concentration of the malondialdehyde standard product is 1 μmol / L, and the specimen treatment solution is 0.5mol / L trichloroacetic acid, the TBA reagent includes 0.1% thiobarbituric acid solution and 0.05mg / L ferric sulfate solution. The kit includes a centrifuge tube with a specification of 0.5-5.0ml, and the malondialdehyde standard, specimen processing solution and TBA reagent are separately packed in bottles.

[0081] In the above example, the preparation steps of malondialdehyde standard product (1umol / L): Accurately weigh 1.64mg of malondialdehyde dimethyl acetal (TMP), add 1000ml of 0.01mol / L hydrochloric acid, stir well at room temperature for 10min, and wait until TMP is fully dissolved Afterwards, aliquot into 0.5ml / bottle and store at 8°C.

[0082] Preparation steps of specimen tre...

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 quantitative determination method and a kit for malondialdehyde content in seminal plasma. The quantitative determination method comprises the following steps of: taking a seminal fluid specimen, centrifuging the seminal fluid specimen, taking a supernatant in a centrifuge tube to obtain a seminal plasma specimen, adding a specimen treatment fluid into the seminal plasmaspecimen to eliminate the interference of proteins and lipids in the seminal fluid specimen, performing uniformly mixing, standing and centrifuging on the product, and drawing the supernatant to obtain a treated specimen; uniformly mixing a malondialdehyde standard substance and a TBA reagent to obtain a standard tube, and uniformly mixing the treated specimen and the TBA reagent to obtain a sample tube, wherein the TBA reagent to comprise a thiobarbituric acid solution and a ferric sulfate solution; reacting the standard tube and the sample tube at 95 DEG C for 30 minutes, cooling the tubes,and measuring the absorbance of the standard tube and the sample tube at 532 nm, 450 nm and 600 nm by using a spectrophotometer; and calculating the measured values through using a formula to obtain the concentration of malondialdehyde in the sample tube; According to the quantitative determination method, the trichloroacetic acid and the TBA reagent of the ferric sulfate solution are added into the seminal plasma specimen, and treatment calculation is conducted by adopting a spectrophotometric method.

Description

technical field [0001] The invention belongs to the technical field of in vitro detection of sperm biochemical indicators, and relates to a quantitative determination method and a kit for the content of malondialdehyde in seminal plasma. Background technique [0002] The etiology of male infertility in modern society has found that the peroxidation of excessive active oxygen free radicals in sperm can lead to the loss of sperm function, which is one of the important reasons for male infertility. Oxygen free radicals are a type of highly active chemical groups produced during the body's oxygen metabolism, which can attack polyunsaturated fatty acids in sperm cell membranes to produce lipid peroxidation products such as malondialdehyde (MDA). Therefore, seminal plasma MDA The content is an indicator of the severity of sperm lipid peroxidation damage. The increase of MDA content in seminal plasma indicates that the lipid peroxidation damage of sperm plasma membrane is strengthe...

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 Applications(China)
IPC IPC(8): G01N21/31
CPCG01N21/31G01N2021/3133
Inventor 张道兵
Owner 珠海高瑞特医疗科技有限公司
Who we serve
  • R&D Engineer
  • R&D Manager
  • IP Professional
Why Patsnap Eureka
  • Industry Leading Data Capabilities
  • Powerful AI technology
  • Patent DNA Extraction
Social media
Patsnap Eureka Blog
Learn More
PatSnap group products

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

© 2024 PatSnap. All rights reserved.Legal|Privacy policy|Modern Slavery Act Transparency Statement|Sitemap|About US| Contact US: help@patsnap.com