Kit for quantitative determination of active oxygen materials in sperms as well as application and using method of kit

A detection kit and oxygen quantification technology, applied in the direction of color/spectral characteristic measurement, measuring device, and analysis through chemical reaction of materials, etc. Convenience and other issues, achieve high sensitivity, overcome imprecise detection, and simple operation

Pending Publication Date: 2018-06-08
BRED LIFE SCI TECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The thiobarbituric acid (TBA) method is not rigorous enough and the results are inaccurate. This method is not convenient for evaluating the situation of active oxygen; the luminol chemiluminescence method is the methodology recommended by the World Health Organization, but it needs to use Strongly acidic luminol is harmful to the operator and the environment; the horseradish peroxidase method requires the use of trichloroacetic acid, an acidic corrosion product, which is also harmful to the operator and the environment, and requires ultrasonic treatment at 0°C. Inconvenient for clinical use

Method used

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  • Kit for quantitative determination of active oxygen materials in sperms as well as application and using method of kit
  • Kit for quantitative determination of active oxygen materials in sperms as well as application and using method of kit
  • Kit for quantitative determination of active oxygen materials in sperms as well as application and using method of kit

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0050]1. Prepare the quantitative detection kit for sperm reactive oxygen species in Example 1, as follows.

[0051] Prepare NBT freeze-dried powder (0.212mmol / L): Accurately weigh 1.2134g of NBT dry powder, make up to 1000mL with purified water, dispense 1mL / bottle, freeze-dry, and add 7mL NBT solution to redissolve when used, that is, the concentration of NBT is 0.212mmol / L.

[0052] Prepare NBT solution (phosphate buffer solution with a concentration of 0.05mol / L): Weigh 2.5857g of potassium dihydrogen phosphate and 29.0093g of disodium hydrogen phosphate dodecahydrate, add an appropriate amount of purified water and stir to dissolve, then dilute the purified water to 1000mL, adjust the phosphate pH to 7.2-7.6.

[0053] Prepare color developer A (0.25mol / L sodium bicarbonate): weigh 21.0025g of sodium bicarbonate, add purified water to the volumetric flask to make up to 1000mL.

[0054] Prepare color developer B (1.62% triethanolamine): take 16.2 mL of triethanolamine, ad...

Embodiment 2

[0084] 1. Preparation of the quantitative detection kit for sperm reactive oxygen species in Example 2, as follows.

[0085] Preparation of freeze-dried powder (0.1mmol / L NBT): Accurately weigh 0.2862g of NBT dry powder, make up to 500mL with purified water, dispense 1mL / bottle, freeze-dry, and add 7mL NBT solution to redissolve before use, that is, the concentration of NBT is 0.1mmol / L.

[0086] Preparation of NBT solution (phosphate buffer solution with a concentration of 0.01mol / L): Weigh 0.25857g of potassium dihydrogen phosphate and 2.90093g of disodium hydrogen phosphate dodecahydrate, add an appropriate amount of purified water and stir to dissolve, then dilute the purified water to 1000mL, use concentrated hydrochloric acid or concentrated sodium hydroxide solution to adjust the pH to 7.2-7.6.

[0087] Prepare color developer A (0.01mol / L sodium bicarbonate): Weigh 0.8401g of sodium bicarbonate, add purified water to the volumetric flask to make up to 1000mL.

[0088...

Embodiment 3

[0118] 1. Preparation of the quantitative detection kit for sperm reactive oxygen species in Example 3, as follows.

[0119] Prepare NBT freeze-dried powder (1mmol / L): Accurately weigh 5.7236g of NBT dry powder, make up to 1000mL with purified water, dispense 1mL / bottle, freeze-dry, and add 7mL NBT solution to redissolve (that is, the concentration of NBT is 1mmol / L.

[0120] Preparation of NBT solution (phosphate buffer solution with a concentration of 0.5mol / L): Weigh 12.9285 g of potassium dihydrogen phosphate and 145.0465 g of disodium hydrogen phosphate dodecahydrate, add an appropriate amount of purified water and stir to dissolve, then dilute the purified water to 1000mL, use concentrated hydrochloric acid or concentrated sodium hydroxide solution to adjust the pH to 7.2-7.6.

[0121] Prepare color developer A (0.3mol / L sodium bicarbonate): weigh 12.6046g sodium bicarbonate, add purified water to the volumetric flask to make up to 500mL.

[0122] Prepare color develop...

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PUM

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Abstract

The invention provides an active oxygen quantitative detection kit. The kit is prepared from Nitro blue tetrazolium chloride monohydrate freeze-dried powder, a blue tetrazolium chloride monohydrate solution, a color developing agent A, a color developing agent B and a standard substance, wherein the chlorinated nitro tetrazole blue freeze-dried powder is powder obtained by freeze-drying blue tetrazolium chloride monohydrate liquid with a concentration of 0.1-1mmol/L; the blue tetrazolium chloride monohydrate solution is a phosphate buffer solution which has a concentration of 0.01-0.5mol/L anda pH value of 7.2-7.6; the color developing agent A is a weak acid buffer solution with a concentration of 0.01-0.3mol/L; the color developing agent B is 0.01-2% triethanolamine solution; the standard substance contains a hydrogen peroxide solution with a known concentration of 0-36mmol/L. The kit provided by the invention has the advantages that the method is scientific, clean and environmentally-friendly, manual operation and automatic batch inspection can be realized, and the kit is simple to operate and easy in clinical promotion.

Description

technical field [0001] The invention relates to a kit for quantitatively detecting reactive oxygen species and its application, and to a method for using the kit to detect the concentration of reactive oxygen species in human sperm in vitro. Background technique [0002] Semen reactive oxygen species (reactive oxygen species, referred to as ROS) substances are mainly produced by the sperm itself and the white blood cells in the semen. In the reproductive system, the antioxidant enzyme system and non-enzymatic antioxidant components keep the production and removal of ROS in a dynamic balance . An appropriate amount of reactive oxygen species plays a variety of roles in sperm function. It is necessary for sperm capacitation and acrosome reaction to maintain the normal function of sperm cells. However, when ROS exists in excess, it will cause cellular oxidative stress and damage the body. The oxidative defense system of the sperm can change the motility function of the sperm, ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N21/01G01N21/78G01N21/31
CPCG01N21/01G01N21/3103G01N21/78
Inventor 胡家纯陶思恩赵毅刘少栩杨红芳程浩
Owner BRED LIFE SCI TECH
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