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A kind of free fatty acid determination kit and preparation method thereof

A free fatty acid and kit technology, applied in biochemical equipment and methods, microbial determination/inspection, etc., can solve problems such as large amount of enzyme used, short storage time, and the activity of enzyme is easily interfered by various factors, Achieve the effect of not easy precipitation, easy filtration and fast filtration

Active Publication Date: 2019-10-11
中拓生物有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] 1) Reagent R2 containing peroxidase and acyl-CoA oxidase has a complex composition and contains a variety of enzymes, whose stability is related to enzyme activity, and enzyme activity is easily interfered by various factors, such as pH, temperature, Ultraviolet rays, heavy metal salts, inhibitors, activators, etc. lead to lack of stability of the kit and short storage time, which affects clinical use;
[0006] 2) The amount of enzyme used in reagent R2 is large, and the reagent is prone to precipitation

Method used

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  • A kind of free fatty acid determination kit and preparation method thereof
  • A kind of free fatty acid determination kit and preparation method thereof
  • A kind of free fatty acid determination kit and preparation method thereof

Examples

Experimental program
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Effect test

Embodiment 1

[0023]

[0024]

[0025] Reagent R1 preparation method: take 800mL of water, add sodium dihydrogen phosphate to 50mmol / L, fully dissolve, adjust the pH to 7.2 with NaOH, add coenzyme A to 0.05mmol / L, ATP to 3mmol / L, ACS to 0.4KU / L, MgCl 2 to 2mmol / L, Trinder substrate to 0.5g / L

[0026] Reagent R2 preparation method: take 800mL of water, add sodium dihydrogen phosphate to 60mmol / L, fully dissolve, adjust pH to 7.2 with NaOH, add FAD to 2mmol / L, 4-AA to 10mmol / L, ACOD to 40KU / L L. Tween 20 to <1%, glycerol to 20%-30%, Brj35 to 1-1.5g / L, EDTA-2Na to 1g / L.

Embodiment 2

[0028]

[0029]

[0030] The preparation method is the same as in Example 1.

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Abstract

The invention relates to the technical field of free fatty acid determining, in particular to a free fatty acid determining kit. The kit comprises a reagent R1 and a reagent R2, wherein the R1 is prepared from coenzyme A, triphosadenine, acyl coenzyme A synthetase, MgCl2, a Trinder substrate and Tween-20, and has the pH of 7.2; the R2 is prepared from flavin adenine dinucleotide, 4 amino-antipyrine, peroxidase, acyl coenzyme A oxidase, Tween-20, glycerinum, 1 to 1.5g / L of brij35-1 and ethylene diamine tetraacetic acid disodium salt and has the pH of 7.2. By adding different combinations of protein protecting agents, the enzyme activity stability is prolonged; the 37-DEG C stability of the enzyme is prolonged from 7 days to 10 days; the enzyme denaturation is slowed down; the reagent R2 cannot easily generate precipitate.

Description

technical field [0001] The invention relates to the technical field of free fatty acid measurement, in particular to a free fatty acid measurement kit, and also to a preparation method of the free fatty acid measurement kit. Background technique [0002] Free fatty acids (NFFA) refer to non-esterified fatty acids or unlipidated fatty acids. The metabolic activity of NEFA in serum is extremely high, and it is easily affected by fat metabolism, glucose metabolism and endocrine metabolism. With the deepening of research and the continuous improvement of technology, the relationship between serum NEFA and diseases has gradually become clear. NEFA has been confirmed to be associated with the occurrence and development of cardiovascular and cerebrovascular, respiratory, digestive, endocrine, immune and other system diseases, as well as tumors and traumatic stress. are closely related to energy metabolism. It is an important substance that induces insulin resistance, and it is a ...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12Q1/48C12Q1/28C12Q1/26
Inventor 解永生梁琳琳隗勇李晓云张强
Owner 中拓生物有限公司
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