Method, reagent and kit for quantitatively detecting free fatty acid

A technology for the quantitative determination of free fatty acids, applied in biological testing, material inspection products, etc., can solve the problems of unsuitable routine testing, high price, and long time consumption, and achieve the effect of fully automated analysis and easy operation

Inactive Publication Date: 2015-06-03
ZHEJIANG KAICHENG BIOTECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] At present, the enzymatic method is generally used to determine the content of free fatty acids in clinical practice, but the existing methods have the disadvantages of poor stability, narrow linear range, cumbersome operation,

Method used

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  • Method, reagent and kit for quantitatively detecting free fatty acid
  • Method, reagent and kit for quantitatively detecting free fatty acid
  • Method, reagent and kit for quantitatively detecting free fatty acid

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0032] Configure the following reagent I and reagent II of the present invention according to the following ingredients and ratios:

[0033] Reagent I:

[0034]

[0035]

[0036] Reagent II:

[0037]

[0038] Mix 300 μl reagent I and 5 μl serum sample in the sample tube, incubate at 37°C for 2-5 minutes, use Hitachi 7180 automatic biochemical analyzer, measure the absorbance A1 at the main wavelength of 546nm and the secondary wavelength of 600nm, and then add to the sample Add 75 μl of reagent II to the medium, mix well, incubate at 37°C for 5 minutes, and measure the absorbance A2 at the same wavelength. Calculate the ΔA sample according to the following formula (1). Use the same method and conditions to measure the absorbance value of the standard solution, and calculate the △A standard. Then calculate the free fatty acid content of serum sample according to formula (2):

[0039] △A=A2-A1 (1)

[0040] Free fatty acid content in serum (mmol / L) = △A sample / △A st...

Embodiment 2

[0043] Configure the following reagent I and reagent II of the present invention according to the following ingredients and ratios:

[0044] Reagent I:

[0045]

[0046]

[0047] Reagent II:

[0048]

[0049] Mix 300 μl reagent I and 5 μl serum sample in a sample tube, incubate at 37°C for 2-5 minutes, use the Abbott C16000 automatic biochemical analyzer, measure the absorbance A1 at the main wavelength of 546nm and the secondary wavelength of 600nm, and then add to the sample Add 75 μl of reagent II to the solution, mix well, incubate at 37°C for 5 minutes, and measure the absorbance A2 at the same wavelength. Calculate the ΔA sample according to the following formula (1). Use the same method and conditions to measure the absorbance value of the standard solution, and calculate the △A standard. Then calculate the free fatty acid content of serum sample according to formula (2):

[0050] △A=A2-A1 (1)

[0051] Free fatty acid content in serum (mmol / L) = △A sample / ...

Embodiment 3

[0053] Configure the following reagent I and reagent II of the present invention according to the following ingredients and ratios:

[0054] Reagent I:

[0055]

[0056]

[0057] Reagent II:

[0058]

[0059]Mix 300 μl reagent I and 5 μl serum sample in a sample tube, incubate at 37°C for 2-5 minutes, use Olympus AU400 automatic biochemical analyzer, measure the absorbance A1 at the main wavelength of 546nm and the secondary wavelength of 600nm, Then add 75 μl of reagent II to the sample, mix well, incubate at 37°C for 5 minutes, and measure the absorbance A2 at the same wavelength. Calculate the ΔA sample according to the following formula (1). Use the same method and conditions to measure the absorbance value of the standard solution, and calculate the △A standard. Then calculate the free fatty acid content of serum sample according to formula (2):

[0060] △A=A2-A1 (1)

[0061] Free fatty acid content in serum (mmol / L) = △A sample / △A standard × standard soluti...

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Abstract

The invention relates to a reagent for quantitatively detecting the content of free fatty acid in human serum. The reagent comprises a reagent I and a reagent II which are respectively placed, wherein the reagent I comprises sodium dihydrogen phosphate, disodium hydrogen phosphate, magnesium chloride, fatty acyl-coenzyme A synthetase, ascorbic acid oxidase, coenzyme A, ATP and 4-aminoantipyrine, and the reagent II contains sodium dihydrogen phosphate, disodium hydrogen phosphate, phenoxy ethanol, fatty acyl-coenzyme A oxidase, peroxidase, chromogen and a surfactant. The kit and the detection method only need dozens of microlitres of serum without centrifugal or electrophoresis and other separation treatments, are simple and convenient to operate, can be used for meeting the requirement of full-automatic analysis, and are suitable for timely accurate detection of large-scale samples.

Description

technical field [0001] The application relates to a quantitative determination method, reagent and kit for free fatty acids in human serum. Background technique [0002] Free fatty acids are substances that neutral fats are broken down into. When glycogen, the energy source for muscle activity, is exhausted, adipose tissue will decompose neutral fat into free fatty acids for energy use. Therefore, free fatty acids can be said to be substances required for sustained activities. [0003] Increased free fatty acids are common in hyperthyroidism, alcohol poisoning, acute myocardial infarction in diabetes, hepatic encephalopathy, long-term fasting, glycogen storage disease and so on. [0004] Reduced seen in hypothyroidism, pituitary insufficiency, Addison's disease. Physiological decrease can be caused after meal. [0005] At present, the enzymatic method is generally used to determine the content of free fatty acids in clinical practice, but the existing methods have the di...

Claims

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Application Information

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IPC IPC(8): G01N33/68
CPCG01N33/92
Inventor 梁朝阳
Owner ZHEJIANG KAICHENG BIOTECH
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