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Lactic acid detection reagent and detection method thereof

A lactic acid detection and reagent technology, applied in the field of chemical detection, can solve the problems of difficulty in simultaneously completing the detection of large quantities of samples, limited instrument throughput, and large demand for samples, and achieves low unit detection costs, simple and easy-to-obtain raw materials, and relevant high coefficient effect

Inactive Publication Date: 2019-11-08
杭州皓阳生物技术有限公司
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AI Technical Summary

Problems solved by technology

Although the instrument detection is highly automated and convenient, it is limited by the throughput of the instrument, it is difficult to complete the detection of a large number of samples at the same time, and it is easily interfered by many coexisting components in the sample
In addition, the use of biochemical analyzers to detect Lactate is limited by the instrument, and supporting reagents, calibration materials and quality control materials are required, and the unit sample cost is relatively high
Although the cost of the ultraviolet method is relatively low, it requires a relatively large amount of sample, which is not conducive to the use of small volume cell culture
The detection cost of the existing technology is relatively high, the required sample volume is large, and a large number of sample detection cannot be completed at the same time, and the use is limited by instruments, consumables, resources, etc.; the market needs a method that can quickly, accurately, and large amounts of lactic acid with a small amount of samples. The detection reagent of content detection, the present invention solves such problem

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  • Lactic acid detection reagent and detection method thereof
  • Lactic acid detection reagent and detection method thereof
  • Lactic acid detection reagent and detection method thereof

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Embodiment Construction

[0040] The present invention will be specifically introduced below in conjunction with the accompanying drawings and specific embodiments.

[0041] Detection principle: Lactate oxidase (LOD) and horseradish peroxidase (HRP) can specifically catalyze L-lactic acid to generate pyruvate and hydrogen peroxide under aerobic conditions, 2'-hydrazine-bis-3 -Ethylbenzothiazoline-6-sulfonic acid (ABTS) is transformed into a soluble green cationic radical under the action of hydrogen peroxide, and the absorbance value can be read using a 430nm spectrum. The higher the lactic acid content in the sample, the more hydrogen peroxide generated after reduction, and the more obvious the color reaction. By setting different concentrations of L-lactic acid standards and samples at the same time to react in a transparent 96-well detection plate, react for 15-20 minutes Afterwards, use a microplate reader to read the absorbance value, and after making a standard curve, you can use the standard cur...

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Abstract

The invention discloses a lactic acid detection reagent and a detection method thereof. The detection reagent includes LOD, HRP, ABTS and a buffer solution; the configuration range of a lactic acid detection reaction system is that the ratio of an ABTS solution to an LOD solution to an HRP solution is (2.5-5):(1.4-2.6):(2-5). The detection method includes the following steps that the absorbance value is read through a 430 nm spectrum, the absorbance value is read through a microplate reader, a standard curve is made, and then the lactic acid content in samples can be calculated out through thestandard curve. The content of a large amount of lactic acid can be detected quickly and accurately by using a small number of samples, and meanwhile, the cost of unit sample detection is reduced.

Description

technical field [0001] The invention relates to the field of chemical detection, in particular to a lactic acid detection reagent and a detection method thereof. Background technique [0002] Lactic acid, an organic acid with formula C 3 h 6 o 3 . In the process of cell culture, lactic acid mainly comes from the incomplete oxidation of glucose in the metabolic process. When the oxidation is insufficient, excessive NADH accumulates, which starts the glycolysis pathway, and lactic acid is produced accordingly. The accumulation of lactic acid will make the culture system The pH decreases, the activity of intracellular enzymes is affected, which limits the normal growth and metabolism of cells, resulting in acidification and even death of cells. In addition, the lye added to adjust the pH and the high concentration of lactic acid itself will lead to an increase in the osmotic pressure of the system, thereby affecting cell growth, protein production, and quality. In the proc...

Claims

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Application Information

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IPC IPC(8): G01N21/31
CPCG01N21/31
Inventor 罗立彦唐慧娴李浩强林帅
Owner 杭州皓阳生物技术有限公司
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