Colorimetric Determination of Pyrophosphate Anion and Its Application to Adenylation Enzyme Assay
Colorimetric Determination of Pyrophosphate Anion and Its Application to Adenylation Enzyme Assay
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DOI：
10.2116/analsci.29.1095
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发表时间：
2013-11-01
影响因子：
1.6
通讯作者：
Hamano, Yoshimitsu
中科院分区：
文献类型：
--
作者：
Katano, Hajime;Watanabe, Hiro;Hamano, Yoshimitsu
本文对乙腈-水混合溶剂中18-溴代焦磷酸（[（P_2O_7）Mo_（18）O_（54）]（4-））阴离子的形成和还原比色法进行了改进。[（P2 O 7）Mo 18 O 54]（4-）阴离子从含有MoO 42-和HCl的乙腈-水溶液中沉淀出来，并重新溶解在纯乙腈或碳酸丙烯酯中。这种分离过程减少了ATP的干扰，并防止了由于过量的Mo（VI）物质而使还原剂抗坏血酸呈黄色。在有机溶剂中，[（P2 O 7）Mo 18 O 54]（4-）阴离子被还原成更强烈的蓝色磷酰焦磷酸盐。本文还介绍了比色法在腺苷酸化酶测定中的应用。
A colorimetric pyrophosphate assay based on the formation and reduction of the 18-molybdopyrophosphate ([(P2O7)Mo18O54](4-)) anion in an acetonittile-water mixed solvent was modified and improved. The [(P2O7)Mo18O54](4-) anion is precipitated from the acetonitrile-water solution containing MoO42- and HCl, and is re-dissolved in neat acetonitrile or propylene carbonate. This separation process decreases the interference by ATP, and prevents a yellow coloration of the reducing agent, ascorbic acid, due to excess Mo(VI) species. In the organic solvent, the [(P2O7)Mo18O54](4-) anion is reduced to a more intense blue molybdopyrophosphate species. The application of the colorimetry to the assay of adenylation enzymes is also described in this note.