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Single stable colorimetric reagent of enzyme in liquid state and its application

A colorimetric and reagent technology, applied in the field of single liquid enzyme colorimetric reagents, can solve the problems of error-prone, crowded, large differences between reagent bottles, etc., and achieve the effects of less error-prone, convenient use and simple programming.

Inactive Publication Date: 2003-10-08
肖洪武
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The disadvantage of dry powder reagents is that they need to be reconstituted when used, the reconstitution time is long and the water quality requirements for reconstitution are high, the difference between reagent bottles after reconstitution by different users is large, the quality cannot be guaranteed, and it is easy to cause waste of reagents. Moreover, the packaging of the dry powder single reagent is not suitable for all kinds of automatic analyzers. It needs to be reconstituted and then transferred to the special bottle of the analyzer.
However, due to the increase of current clinical testing items, the reagent positions of the analytical instruments have become crowded. As the amount of tested specimens tends to increase, the frequency of reagent replacement is high, and errors are prone to occur. Dual reagents are measured in two steps, which takes a long time. Due to each The measurement items are divided into two kinds of reagents. It is easy to cause confusion when loading reagents, causing confusion in measurement results, wasting reagents, and affecting work quality. Some users who use small and medium-sized instruments have insufficient reagent storage space.

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0012] Comparative example 1

[0013] Use a new water-soluble developer to replace the previous phenol or chlorophenol: such as TOPS, TOOS, TODB, ADPS, HDAOS, ADOS, DHBS, TBHBA, N-ethyl-(βaminoethyl) m-aniline, etc., the dosage is 0.1 ~50mmol / L. The novel water-soluble developer of the present invention can also be used in combination with conventional phenol, p-hydroxybenzoic acid, 4-chlorophenol, 2,4-dichlorophenol, etc. in appropriate amounts. Most of the new water-soluble chromogenic reagents are purple or blue in color and avoid the absorption peak of hemoglobin, which can effectively reduce the interference of hemoglobin (red) in the specimen, and the dual-wavelength measurement at 600nm / 700nm can reduce the interference of lipid turbidity, and there is no Corrosiveness of phenol.

[0014] Comparative example 2

[0015] The test results show that the new water-soluble chromogenic agent used in the present invention can greatly reduce the damage t...

Embodiment 3

[0016] Use different surfactants to promote the dissolution of the protein in the specimen to release the substance to be tested, and reduce the interference caused by the high fat content of the specimen. Nonionic surfactants, amphoteric surfactants, and polyethers can be used alone or in combination, and the dosage is 0.05~ 100g / L

[0017] Example 2

[0018] Result of the test shows that embodiment 3 of the present invention does not use sodium cholate, uses nonionic surfactant alone to have high stability equally, and its advantage: comparative example 2, embodiment 2 reagent contain sodium cholate and easily interfere other clinical measurement items such as bile Acid, high-density and low-density cholesterol are directly measured, but Example 3 does not have this drawback. Example 4:

Embodiment 4

[0019] This example provides how to improve enzyme stability in enzyme colorimetric reagents.

[0020] Usually, enzymes are not very stable in liquid state, which can easily cause enzyme inactivation, protein precipitation, and reagent blank rise, while polyols, albumin, amino acids, monosaccharides, disaccharides, chelating agents, antioxidants, preservatives, protease inhibitors are all It can enhance its stabilizing effect and can be used alone or in combination.

[0021] Polyols below six carbons not only have a stabilizing effect but also have an antifreeze effect (high temperature and freezing can damage enzyme activity): common ones are glycerin, ethylene glycol, propylene glycol, mannitol, diethylene glycol, etc., and the dosage is 1 to 500g / L.

[0022] Proteins can interact with enzyme molecules to prevent dissociation into subunits or denaturation and inactivation tendency: bovine serum albumin (with certain antioxidant properties), human serum albumin, ovalbumin, ...

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Abstract

A high-stability liquid chromometric enzyme reagent is prepared from buffering liquid, water-soluble chromogenic agent, 4-aminoantipyrin, peroxidase, oxidase for generating hydrogen peroxide and enzyme stabilizer. Its advantages are durable stability, and high efficiency and correctness.

Description

technical field [0001] The invention relates to an analysis reagent and its application, in particular to a stable single liquid enzyme colorimetric reagent and its application. technical background [0002] Nowadays, enzyme colorimetric assay reagents are used clinically. Common assay reagents include triglyceride, cholesterol, glucose, uric acid, creatinine and other assay reagents. The principle is that almost all the compounds to be detected generate hydrogen peroxide through specific oxidases for colorimetry. Determination. [0003] In the past, the determination reagents of enzyme colorimetry were mainly dry powder reagents. The disadvantage of dry powder reagents is that they need to be reconstituted when used, the reconstitution time is long and the water quality requirements for reconstitution are high, the difference between reagent bottles after reconstitution by different users is large, the quality cannot be guaranteed, and it is easy to cause waste of reagents...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/26C12Q1/54C12Q1/60C12Q1/61C12Q1/62
Inventor 肖洪武
Owner 肖洪武
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