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Stable p-nitrophenyl phosphate solution and preparation method thereof

A technology of p-nitrophenylphosphoric acid and nitrophenylphosphoric acid, which is applied in the direction of biochemical equipment and methods, microbiological measurement/inspection, etc., can solve problems such as complicated operation steps, and achieve simple process, simplified production process, and good correlation Effect

Active Publication Date: 2018-05-08
ZHONGSHAN CHUANGYI BIOCHEM ENG
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Problems solved by technology

Patent US4132598 mentions that the use of phenols or compounds containing imidazole structures can be used as a stabilizer for PNPP in solution. After mixing under certain conditions, dissolve it in deionized water, and then adjust the pH of the solution to about 8.6. The operation steps are more complicated

Method used

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  • Stable p-nitrophenyl phosphate solution and preparation method thereof
  • Stable p-nitrophenyl phosphate solution and preparation method thereof
  • Stable p-nitrophenyl phosphate solution and preparation method thereof

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Embodiment

[0017] 1. Preparation process

[0018] The alkaline phosphatase kit consists of two parts: reaction buffer and substrate storage solution, wherein the alkaline phosphatase substrate is stored

[0019] Preparation method of stock solution:

[0020] 1) Dissolve Trisbase and imidazole in deionized water successively;

[0021] 2) Adjust the pH of the above solution to 8.4 with hydrochloric acid (1.0mol / L) solution, then add sodium azide, and stir to fully dissolve;

[0022] 3) Add disodium p-nitrophenyl phosphate, mix thoroughly and then settle to volume;

[0023] 4) Transfer the prepared storage solution into a brown reagent bottle and store it in the dark at 2-8°C.

[0024] 2. Relevant formula

[0025] Prepare alkaline phosphatase substrate stock solution according to the following recipe:

[0026]

[0027] Prepare the reaction buffer according to the following recipe

[0028]

[0029]

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Abstract

The invention discloses a stable p-nitrophenyl phosphate solution. The solution contains a Trisbase-HCl buffer solution, imidazole, p-nitrophenyl phosphate disodium and sodium azide. The preparation method comprises the following steps: 1) sequentially dissolving Trisbase and the imidazole into deionized water; 2) adjusting the pH value of the solution to 8.4 with a hydrochloric acid solution, adding the sodium azide and stirring to completely dissolve the sodium azide; 3) adding the p-nitrophenyl phosphate disodium, mixing completely and uniformly and fixing volume; and 4) transferring the prepared storage liquid into a brown reagent bottle and keeping in a dark place at 2 to 8 DEG C. The p-nitrophenyl phosphate solution has high stability, the storage liquid has high precision degree andrelativity when being applied to an alkaline phosphatase measuring kit, and the product has high stability.

Description

technical field [0001] The invention relates to the field of biological detection, in particular to a stable p-nitrophenyl phosphoric acid solution and a preparation method thereof. Background technique [0002] Alkaline phosphatase (ALP or AKP) is an enzyme that is widely distributed in tissues such as the human liver, bone, intestine, kidney and placenta, and is excreted from the liver to the bile. This enzyme can catalyze the removal of the 5' phosphate group from nucleic acid molecules, thereby converting the 5'-P end of the DNA or RNA fragment into a 5'-OH end. Alkaline phosphatase is mainly used for the examination of obstructive jaundice, primary liver cancer, secondary liver cancer, and cholestatic hepatitis. When suffering from these diseases, the liver cells over-produce ALP, which enters the blood through the lymphatic channel and hepatic sinusoids, and at the same time, due to the obstruction of bile excretion in the intrahepatic biliary tract, it flows back int...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/42
CPCC12Q1/42
Inventor 崔海林黄小全李冰曾晓君黄清媚
Owner ZHONGSHAN CHUANGYI BIOCHEM ENG
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