ATP content determination kit and method thereof

Abstract

The invention discloses an ATP content determination kit and a method thereof. The kit comprises a reagent I prepared by dissolving concentrated perchloric acid in distilled water, a reagent II prepared by dissolving sodium hydroxide in distilled water, a reagent III prepared from sodium phosphate dibasic dodecahydrate, a reagent IV prepared from sodium dihydrogen phosphate dihydrate and a reagent V prepared from ATP. The method improves the extraction process of ATP and is capable of removing influence of impurities in a sample by firstly adding an acidic extraction solution into the sample for fully extracting and then adding an alkaline extraction solution for neutralizing. The minimum detection limit of the method reaches up to 0.1Mu mol / mL; compared with a conventional spectrophotometric method, the minimum detection limit is significantly improved. The reagent components of the kit only comprise absolute methanol, disodium hydrogen phosphate and sodium dihydrogen phosphate; the detection cost is greatly reduced. The method is qualitative and quantitative in through-peak area and retention time; the detection accuracy is significantly improved.

Description

technical field [0001] The invention belongs to the field of life sciences and relates to a kit, in particular to a kit for measuring ATP content and a method thereof. Background technique [0002] ATP widely exists in animals, plants, microorganisms and cultured cells, and is the currency of biological energy. Energy charge is the main parameter describing the energy metabolism state of cells. Determination of ATP content and calculation of energy charge can reflect the state of energy metabolism. [0003] At present, the existing method for detecting ATP content in the market is the spectrophotometric method based on the colorimetry of phosphomolybdic acid. The method utilizes creatine kinase to catalyze the reaction of creatine and ATP to generate phosphocreatine, and detects the content of phosphocreatine at 700 nm to reflect the content of ATP. But this method also has the following disadvantages: [0004] 1. The detection sensitivity is low, and the minimum detectio...

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Problems solved by technology

[0004] 1. The detection sensitivity is low, and the minimum detection limit is 1 μmol / mL

[0005] 2. Susceptible to the interference of impurities in the sample, affecting the detection accuracy

[0006] 3. The reaction system is greatly affected by temperature and pH value, resulting in poor stability and poor repeatability of the measurement results

[0007] 4. The creatine kinase reagent that needs to be used is expensive and the cost is high