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Method for measuring diethyl p-nitrophenyl phosphate esterase active concentration and diagnostic kit

A technology of nitrophenyl phosphatase and diagnostic kit, which is applied in the field of determination of diethyl-p-nitrophenyl phosphatase activity concentration and diagnostic kit, and can solve the problems of decreased antioxidant effect, heart disease or stroke, activity Low concentration and other problems, to achieve the effect of high accuracy, simple method and high precision

Inactive Publication Date: 2007-06-06
SUZHOU ANJ BIOTECHNOLOGY CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0003] The active concentration of diethyl-p-nitrophenyl phosphatase in smokers is lower than that of non-smokers or ex-smokers, so the antioxidant effect is also reduced, resulting in more cholesterol deposition in blood vessels, and excess Cholesterol can clog blood vessels, eventually leading to heart attack or stroke

Method used

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  • Method for measuring diethyl p-nitrophenyl phosphate esterase active concentration and diagnostic kit
  • Method for measuring diethyl p-nitrophenyl phosphate esterase active concentration and diagnostic kit
  • Method for measuring diethyl p-nitrophenyl phosphate esterase active concentration and diagnostic kit

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Experimental program
Comparison scheme
Effect test

Embodiment 1

[0043] Embodiment one (single dose)

[0044] Prepare the diethyl-p-nitrophenyl phosphatase diagnostic kit according to the following ingredients and dosage:

[0045] Tris(carboxymethyl)aminomethane-hydrochloric acid buffer 100mmol / L,

[0046] Ammonium sulfate 80g / ml,

[0047] NADH 0.25mmol / L,

[0048] Aldehyde dehydrogenase 8000U / L,

[0049] Phenyl acetate 16mmol / L.

[0050] After the reagents are all dissolved and prepared, they are divided into bottles and freeze-dried to make dry powder reagents; before use, add purified water and reconstitute for use.

[0051] Set on the automatic biochemical analyzer: temperature 37°C, reaction time 10 minutes, initial absorbance 1.8±0.7, test main wavelength 340nm, test subwavelength 405nm, tested diethyl-p-nitrophenyl phosphatase sample and The volume ratio of reagents is 1:25, the reaction direction is negative reaction (falling reaction), the delay time is about 1 minute, the detection time is about 2 minutes, and the theoretical K...

Embodiment 2

[0053] Embodiment two (two doses)

[0054] Prepare the diethyl-p-nitrophenyl phosphatase diagnostic kit according to the following ingredients and dosage:

[0055] Reagent I——

[0056] Tris(carboxymethyl)aminomethane-hydrochloric acid buffer 100mmol / L,

[0057] Glycerol 2mol / L,

[0058] thio-NADH 0.25mmol / L,

[0059] Phenyl acetate 12mmol / L;

[0060] Reagent II——

[0061] Tris(carboxymethyl)aminomethane-hydrochloric acid buffer 100mmol / L,

[0062] Propylene Glycol 50% v / v,

[0063] Aldehyde dehydrogenase 12000U / L.

[0064] After the reagents are all dissolved and prepared, they are divided into bottles to make liquid double reagents, which can be used directly.

[0065] Set on the automatic biochemical analyzer: temperature 37°C, reaction time 10 minutes, initial absorbance 1.8±0.7, test main wavelength 340nm, test subwavelength 405nm, tested diethyl-p-nitrophenyl phosphatase sample and The volume ratio of reagent I and reagent II is 2:20:5, the reaction direction is ...

Embodiment 3

[0067] Embodiment three (three doses)

[0068] Prepare the diethyl-p-nitrophenyl phosphatase diagnostic kit according to the following ingredients and dosage:

[0069] Reagent I——

[0070] Tris(carboxymethyl)aminomethane-hydrochloric acid buffer 100mmol / L,

[0071] Ammonium sulfate 80g / ml,

[0072] NADPH 0.25mmol / L;

[0073] Reagent II——

[0074] Tris(carboxymethyl)aminomethane-hydrochloric acid buffer 100mmol / L,

[0075] Ethylene glycol 2mol / L,

[0076] Phenyl acetate 20mmol / L;

[0077] Reagent III——

[0078] Tris(carboxymethyl)aminomethane-hydrochloric acid buffer 100mmol / L,

[0079] Glycerin 50% v / v,

[0080] Aldehyde dehydrogenase 6000U / L.

[0081] After all the reagents are dissolved and prepared, they are divided into bottles to make liquid three reagents, which can be used directly.

[0082] When measuring the activity concentration of diethyl-p-nitrophenyl phosphatase, set on the automatic biochemical analyzer: temperature 37°C, reaction time 10 minutes, ini...

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Abstract

This invention disclosed a determination method for paraoxonase activity and the reagent box. The procedures are as follows: react with phenylacetate to produce acetic acid, oxidate the deoxidize style coenzyme to coenzyme, measure the coenzyme at 340nm absorbency, calculate the activity and concentration. The reagent box contains buffer, deoxidize style coenzyme, phenylacetate and stabilizer. The result obtained by this method was precise and accurate.

Description

technical field [0001] The invention relates to the detection of the activity concentration of diethyl-p-nitrophenyl phosphatase in the fields of medicine, food, environment, etc., and in particular to the determination of diethyl-p-nitrophenyl phosphate by using an enzyme colorimetric method and an enzyme (coupling) method The method for esterase activity concentration and the diethyl-p-nitrophenyl phosphatase diagnostic kit prepared thereby belong to the technical field of analysis and detection of diethyl-p-nitrophenyl phosphatase activity concentration. Background technique [0002] Diethyl-p-nitrophenyl phosphatase in serum is mainly expressed in the liver, and its function is to hydrolyze phosphate in tissues. Diethyl-p-nitrophenyl phosphatase in serum is related to the metabolism of high and low density lipoproteins in the human body. Diethyl-p-nitrophenyl phosphatase can protect low-density lipoproteins from oxidative modification, so it can be considered that dieth...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N21/31C12Q1/26C12Q1/00
Inventor 王尔中
Owner SUZHOU ANJ BIOTECHNOLOGY CO LTD
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