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Two-step enzyme testing method of triglycercide in blood serum

A technology of triglyceride and determination method, which is applied in the direction of material analysis by observing the influence of chemical indicators, and analysis by chemical reaction of materials, etc., can solve problems such as endogenous glycerol interference, and achieve economical, convenient and easy-to-use High performance, high accuracy, and the effect of increasing the cost of reagents

Inactive Publication Date: 2010-06-30
WHITMAN BIOTECH NANJING
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0008] In order to solve the problem of the interference of endogenous glycerol in the determination method of TG in serum in the prior art, the present invention provides an economical, convenient, high-accuracy, and capable of eliminating the influence of endogenous glycerol in serum triglycerides. step-by-step enzyme assay

Method used

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  • Two-step enzyme testing method of triglycercide in blood serum
  • Two-step enzyme testing method of triglycercide in blood serum
  • Two-step enzyme testing method of triglycercide in blood serum

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0021] Composition of reagents:

[0022] a. Reagent I:

[0023] Each liter of Tris-HCl buffer contains Tris 0.15mmol (pH7.6), 2,4-dichlorophenol 3.0 mmol, MgSO 4 12mmol, sodium cholate 4.2mmol, Triton X-100 0.12g, ATP 1.2mmol, GK 600U, GPO 3600U, POD1200U, 4-aminoantipyrine 1.6mmol, Proclin-300 preservative 200μl.

[0024] b. Reagent II:

[0025] Each liter of Tris-HCl buffer contains Tris 0.15 mmol (pH7.6), LPL 24000U, Proclin-300 preservative 200μl.

[0026] c. Standard solution: 2mmol / L trioleate aqueous solution.

[0027] Among them, MgSO 4 It is an activator of glycerol kinase, Triton X-100 is a surfactant, Proclin-300 is a liquid high-efficiency preservative, and sodium cholate is a bacteriostatic agent.

Embodiment 2

[0029] In reagent I, 2,4-dichlorophenol was changed to 4-chlorophenol. Content unchanged. All other ingredients remain unchanged, and reagent II remains unchanged.

Embodiment 3

[0031] Measurement procedure

[0032] Two-reagent method: On the Japanese OLYMPUS AU2700 fully automated biochemical analyzer, the instrument automatically adds 2 µm of sample to 150 µm of Reagent I and mixes evenly, incubates at 37°C for 3 minutes, adds 50 µm of Reagent II, mixes evenly, and incubates at 37°C for 5.1 minutes , Automatic analyzer detects at 500nm wavelength. The instrument automatically calculates the TG result. See Table 1 for details

[0033] Table 1. Automatic biochemical analyzer test conditions of the present invention

[0034]

[0035] response OD TG Calculated value = OD 2 -OD 1 ×[(SV+R 1 V 1 ) / (SV+R 1 V 1 +R 2 V 2 )]

[0036] Triglyceride concentration = F × OD TG

[0037] where OD TG is the absorbance produced by triglycerides. OD 1 Is the sample added to the reagent? Absorbance measured after reaction, OD 2 Is the sample added to the reagent? Absorbance measured after reaction, SV is the volume of serum sample, R 1 V 1 Is it ...

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Abstract

The invention discloses a two-step enzyme testing method of triglycercide in blood serum, belonging to a method for testing material according to the color change of the result of test reaction with visible light. The method has the technical scheme as follows: double-color raw materials are in reagent I, and reagent II only has active constituent of lipoprotein lipase. The test method comprises the steps of: bathing the blood serum and the reagent I under the temperature of 37 DEG C for 3-5 minutes; reaching free glycerin in the blood serum with the reagent I to generate quinone imide; bathing under the temperature of 37 DEG C for 4-7 minutes after adding the reagent II; and hydrolyzing the triglycercide and reacting to generate the red quinone imide. An instrument is used for detecting at the position with the wave length of 500-520nm, the quinone imide generated by the reaction of the reagent I is taken as blank, and the content of the triglycercide is computed through the quinone imide generated by the reaction of the reagent II. The method is not influenced by endogenic glycerine, has the same steps and range as the existing two-step method, does not increase the cost of the reagent, and is economic and convenient, thereby being a triglycercide testing method with higher accuracy.

Description

technical field [0001] The present invention belongs to a kind of determination method that comprises enzyme; Or utilize visible light, produce the method for testing material through the color change of test reaction result, especially relate to a kind of two-step enzyme determination that uses biochemical analyzer to detect triglyceride in serum method. Background technique [0002] The determination methods of triglyceride (TG) in serum can be generally divided into three categories: chemical method, enzymatic method and chromatographic method. Early methods of determination were estimated as the difference between total lipid and cholesterol and phospholipids. The chemical method extracts TG in the specimen with an organic solvent, removes phospholipids and other interfering substances in the extract, hydrolyzes (saponifies) TG with alkali, oxidizes glycerol with periodic acid to form formaldehyde, and then uses color reaction to measure formaldehyde. More accurate is ...

Claims

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Application Information

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IPC IPC(8): G01N21/78
Inventor 李立和
Owner WHITMAN BIOTECH NANJING
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