A NEW AND CONVENIENT COLORIMETRIC DETERMINATION OF INORGANIC ORTHO-PHOSPHATE AND ITS APPLICATION TO THE ASSAY OF INORGANIC PYROPHOSPHATASE
A NEW AND CONVENIENT COLORIMETRIC DETERMINATION OF INORGANIC ORTHO-PHOSPHATE AND ITS APPLICATION TO THE ASSAY OF INORGANIC PYROPHOSPHATASE
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DOI：
10.1016/0003-2697(81)90082-8
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发表时间：
1981-01-01
影响因子：
2.9
通讯作者：
LAHTI, RJ
中科院分区：
文献类型：
--
作者：
HEINONEN, JK;LAHTI, RJ
建立了一种新的简便的测定磷的方法。通过将整个测定混合物溶解在丙酮中比色测量磷钼酸盐，而不还原为钼蓝，其中磷钼酸盐为亮黄色。酸不稳定磷酸盐的水解（例如，磷酸肌酸）不会引起问题，因为额外的肌酸在颜色显现后立即与柠檬酸盐络合。如果存在高浓度的强还原剂和SH化合物，则通过加入H2 O2消除。洗涤剂、有机碱、蛋白质和蔗糖不干扰。该测定法与Fiske-SubbaRow方法的大多数改进方法一样灵敏。在常规程序中，有用的范围是50-1500 nmol的Pi。本文介绍了该方法在大肠杆菌无机焦磷酸酶测定中的应用。
A new and convenient method for the determination of Pi was developed. Phosphomolybdate is measured colorimetrically, without reduction to molybdenum blue, by dissolving the whole assay mixture in acetone, where phosphomolybdate is bright yellow. The hydrolysis of acid-labile phosphates (e.g., creatine phosphate) causes no problems, because extra molybdate is complexed with citrate immediately after the color has been developed. Strong reductants and SH compounds which interfere, if present in high concentrations, are eliminated by adding H2O2. Detergents, organic bases, protein and sucrose do not interfere. The assay is as sensitive as most modifications of the Fiske-SubbaRow method. In the routine procedure the useful range is 50-1500 nmol of Pi. The application of the method to the assay of inorganic pyrophosphatase in the cells of Escherichia coli is described.