Determination reagent for cholesterol in high density lipoprotein and preparing method thereof

A high-density lipoprotein and cholesterol technology, applied in the fields of medicine and biochemistry, can solve the problem that the specificity is not necessarily good, and achieve the effects of good specificity, lower detection costs, and reduced detection workload.

Active Publication Date: 2009-12-02
浙江伊利康生物技术有限公司
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  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

And the specificity is not necessarily good

Method used

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  • Determination reagent for cholesterol in high density lipoprotein and preparing method thereof
  • Determination reagent for cholesterol in high density lipoprotein and preparing method thereof
  • Determination reagent for cholesterol in high density lipoprotein and preparing method thereof

Examples

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preparation example Construction

[0042] Two, the preparation of high-affinity cholesterolase compound:

[0043] 1. Activate one or more mixtures of the above compounds in aqueous medium

[0044]One or more mixtures of the above-mentioned compounds are activated in an aqueous medium by the known azide method, carbodiimide method, succinimide method or periodate oxidation method, and the selected method is preferably able to Maximum maintenance of the activity of the prepared enzyme compound. The activation conditions are: the molar ratio of the compound to the activator is 1:1-1:10, the concentration of the compound is 1-50 mg / ml, the temperature is 0-25°C, the pH is in the range of 4-11, and the time is 0.1-24 hours . As we all know, the activators are azide, p-cyclohexyl-2-(4-morpholine) in the azide method, carbodiimide method, succinimide method or periodate oxidation method, respectively. ) ethyl-carbodiimide-methyl-p-toluenesulfonate, N-hydroxysuccinimide and sodium periodate. Some of the compounds d...

Embodiment 1

[0060] Example 1: The high-affinity enzyme cholesterol esterase compound and the high-affinity enzyme cholesterol oxidase compound in this example are prepared from the agave steroid glycoside and enzyme in the steroid glycoside compound; the configuration method is as described above, in:

[0061] The activation conditions for compound activation are:

[0062] The molar ratio of the compound to the activator is 1:1, the temperature is 15° C., the pH value is 8, and the time is 10 hours.

[0063] The molar ratio of the buffer solution is 100mmol / L, and the pH value of the buffer solution is 7.

[0064] The activated compound was mixed with the dissolved enzyme, and the solution was reacted at a temperature of 15° C. for 20 hours.

[0065] The agave glycoside is pre-dissolved in the following organic solvent mixture: a mixture of formyl dimethylamine and toluene at a ratio of 2:1, and the weight ratio of the compound to the organic solvent mixture is 1:0.01.

[0066] Reagent...

Embodiment 2

[0088] In this embodiment, cholesterol esterase compound and cholesterol oxidase compound are used to prepare from tea saponin and enzyme in triterpene glycoside compounds; the configuration method is as described above, wherein:

[0089] The activation conditions for compound activation are:

[0090] The molar ratio of the compound to the activator is 1:5, the temperature is 0° C., the pH value is 4, and the time is 24 hours.

[0091] The molar ratio of the buffer solution is 50mmol / L, and the pH value of the buffer solution is 9.

[0092] The activated compound is mixed with the dissolved enzyme, and the solution is reacted at 0°C for 24 hours;

[0093] The tea saponin is predissolved in the following organic solvent mixture: a mixture of formyl dimethylamine and ethanol at a ratio of 2:1; the weight ratio of the compound to the organic solvent mixture is 1:15.

[0094] Reagent I

[0095] The dosage of cholesterol esterase compound is 5000U / L

[0096] The dosage of chole...

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Abstract

The invention relates to a reagent for measuring high-density lipoprotein cholesterol in serum and a preparation method. The technical problem to be solved by the present invention is to provide a method for preparing a reagent for measuring cholesterol in high-density lipoprotein and the improvement of the reagent. The reagent prepared by this method should have good specificity, simple and convenient detection, high test accuracy, and It can be detected on various types of automatic biochemical analyzers. The technical scheme is: the components of the reagent are as follows: A. high-affinity enzyme compound, which is obtained by the reaction of enzyme and compound, and the molar mixing ratio of the two components when reacting is 1:1-20:1; B. surfactant, The mixing ratio is 0.003-30% of the weight percentage of component A; C. appropriate amount of preservatives, stabilizers and chromogens. An appropriate amount of catalyst and anti-interference substances are also added to the reagent. The preparation method comprises the following steps: 1. compound activation; 2. enzyme dissolution; 3. mixed reaction; 4. configuration.

Description

technical field [0001] The invention relates to a preparation method and a reagent for measuring serum components, in particular to a preparation method and a reagent for measuring high-density lipoprotein cholesterol in serum, which can be widely used in the technical fields of medicine and biochemistry. Background technique [0002] The incidence of cardiovascular and cerebrovascular diseases is negatively correlated with serum high-density lipoprotein cholesterol (HDL-C), which has been confirmed by epidemiology and related clinics. The detection of high-density lipoprotein cholesterol can also be widely used in relevant clinical diagnosis and treatment, and as a lipid One of the risk indicators of metabolic abnormalities. Therefore, the accurate determination of high-density lipoprotein cholesterol is very important, especially at the lower limit of high-density lipoprotein cholesterol, a very small error may cause a relatively large impact, and the result of high-densit...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12Q1/60C12Q1/44C12Q1/26G01N21/78
Inventor 王贤理蒙凯
Owner 浙江伊利康生物技术有限公司
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