Alpha-oxoglutarate content measurement kit and measurement method thereof

Abstract

An Alpha-oxoglutarate content measurement kit comprises a box main body, a cover plate, a rotation shaft and a fixed seat. A detection method comprises the steps of sample preparation, in which a sample is taken, PH7.8 phosphate buffer liquid is added for grinding, centrifugation is performed for 10 minutes, a supernatant liquid is waited for measurement; detection step, in which the detection step comprises the steps of 1, pre-heating by a photometer, and performing zero setting with distilled water; 2, adding a standard point, alanine, NAD+, glutamate dehydrogenase, 2-(4-iodobenzene)-3-(4-nitrobenzene)-5-(thiobenzoate)-2H-tetrazolium salt, a phosphate buffer liquid and alanine aminotransferase into a standard measurement pipe, adding alanine, NAD+, glutamate dehydrogenase, 2-(4-iodobenzene)-3-(4-nitrobenzene)-5-(thiobenzoate)-2H-tetrazolium salt, a phosphate buffer liquid and alanine transaminase into a blank pipe, adding the to-be-measured supernatant liquid and then adding constituents in the blank pipe into a sample measurement pipe, and adding the to-be-measured supernatant liquid, NAD+, glutamate dehydrogenase, L2-(4-iodobenzene)-3-(4-nitrobenzene)-5-(thiobenzoate)-2H-tetrazolium salt and a phosphate buffer liquid into a contrast pipe; and 3, immediately and uniformly mixing, reacting for 30 min, transferring to a glass cell, reading a light absorption value A of each pipe, and calculating Alpha-oxoglutarate(Alpha-KG) content according to sample mass. The method is simple and easy to operate.

Description

technical field [0001] The invention relates to the field of detection of α-ketoglutarate content, in particular to a method for determining the content of α-ketoglutarate. Background technique [0002] At present, with the advancement of science and technology, the determination of α-ketoglutaric acid content has been more and more applied to many aspects, and the domestic method of detecting α-ketoglutaric acid content is generally using high performance liquid chromatography. (HPLC) detection, this method requires expensive detectors, requires experimenters to have relevant instrument operation knowledge and experience, and it takes a long time from preparing samples to preparing test solutions for on-machine detection and requires a large amount of organic reagents. The cost of instruments, consumables and reagents used in this method is high, and there are certain professional requirements for instrument operators. It is difficult for general laboratories to implement, ...

AI Technical Summary

Problems solved by technology

[0002] At present, with the advancement of science and technology, the determination of α-ketoglutaric acid content has been more and more applied to many aspects, and the domestic method of detecting α-ketoglutaric acid content is generally using high performance liquid chromatography. (HPLC) detection, this method requires expensive detectors, requires experimenters to have relevant instrument operation knowledge and experience, and it takes a long time from preparing samples to preparing the liquid to be tested on the machine and requires a large amount of organic reagents

The cost of instruments, consumables and reagents used in this method is relatively high, and there are certain professional requirements for instrument operators. It is difficult for general laboratories to implement, especially for the experimental screening of large batches of samples, both in terms of time and reagent cost.

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