Detection of related substances of beta-nicotinamide adenine dinucleotide phosphate disodium salt_Industrial additives

Background and overview^[1]

Beta-nicotinamide adenine dinucleotide phosphate disodium salt is usually obtained by hydrolyzing β-nicotinamide adenine dinucleotide phosphate NADP into salt. NADP is a coenzyme. It is a substance that combines β-nicotinic acid amide adenine dinucleotide NAD with a phosphate molecule through an ester bond. It is widely used in the energy metabolism process of organisms. Purine nucleotides are important energy substances in the body. β-Nicotinamide adenine dinucleotide NAD and β-nicotinamide adenine dinucleotide phosphate NADP are coenzymes necessary for cellular energy metabolism and play a central role in energy metabolism. In addition, purine nucleotides also play an important transfer role in biological processes. Protons in energy reactions are usually transferred to NAD and NADP first, and then reduced to NADH and NADPH, and then transferred to oxygen through electrons to release energy. The generated reduced NADH and NADPH are coenzymes of glutathione reductase and have important antioxidant functions.

Detection^[1]

A method for determining the content of β-NADP disodium salt and related substances. The related substances are NAD, ATP, and ADP, which is characterized by including the following steps:

A. Take the sigma reference substance: beta calcium phosphate a-nicotinamide adenine dinucleotide phosphate disodium salt is dissolved with diluent into a reference substance solution with a concentration of 1 mg/ml; take beta-nicotinamide adenine dinucleotide Dissolve the nucleotide disodium phosphate salt sample with diluent into a test solution with a concentration of 0.5mg/ml;

B. Take 20ul each of the reference solution and the test solution, inject them into the liquid chromatograph, record the peak area of ​​beta-nicotinamide adenine dinucleotide phosphate disodium salt in the chromatogram, and follow the external standard Method, use anhydrous beta-nicotinamide adenine dinucleotide phosphate disodium salt to calculate the HPLC content, not less than 98%;

C. Dissolve the sigma reference substances NAD, ATP, and ADP into impurity reference substance solutions with concentrations of 5ug/ml, 2.5ug/ml, and 2.5ug/ml respectively in the mobile phase. Take beta-nicotinamide adenine di Dissolve the nucleotide disodium phosphate salt sample with mobile phase into a sample solution with a concentration of 0.5mg/ml;

D. Take 20ul of each impurity reference solution of NAD, ATP and ADP, inject it into the liquid chromatograph, record the peak areas of NAD, ATP and ADP in the chromatogram, and then calculate beta-nicotinamide according to the external standard method The contents of NAD, ADP, and ATP in the adenine dinucleotide phosphate disodium salt sample, where NAD, ADP, and ATP do not exceed 1.0%, 0.5%, and 0.5% respectively.