Assessment of ionic quenching on soil ATP bioluminescence reaction
Assessment of ionic quenching on soil ATP bioluminescence reaction
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DOI：
10.1016/s0038-0717(00)00104-8
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发表时间：
2001-01-01
影响因子：
9.7
通讯作者：
Chikushi, J
中科院分区：
文献类型：
--
作者：
Wen, G;Voroney, RP;Chikushi, J
当检测ATP时，从土壤中提取的带有ATP的go离子可能会干扰ATP荧光素-荧光素酶的发光反应。研究了土壤中可提取阳离子和阴离子的典型浓度范围。使用商用ATP测定产品（Sigma Chemical Co.）。当Cu2+和Zn2+浓度为0.10 mM， Ca2+浓度为1.00 mM时，淬灭效果明显。在1.00 mM处淬火顺序为：Cu2+ > Zn2+ Ca2+ = Na+ = Mg2+, Mg2+ = Mn2+， Ca2+和Na+ > Mn2+。选用特殊的重金属阳离子，其淬火效果为Ti3+ > Hg2+ > Cr3+。由于EDTA的加入可以缓解阳离子猝灭，因此对三种形式的EDTA （Mg、Na和酸性EDTA）进行了测试。发现Mg-EDTA优于其他两种。0.01和0.05 mM浓度的PO43-和0.01和0.10 mM浓度的NO3-显著提高了ATP的发光率（8-13%）。然而，相同浓度的SO42-显著降低了光发射。CO32-和Cl-的猝灭仅在高浓度（3.20 mM及以上）下观察到。在6.4 mM浓度下，阴离子的猝灭顺序为：PO43- > CO32- > SO42- > NO3- > Cl-。离子诱导的光发射增强或减弱会导致土壤ATP的显著高估或低估。虽然添加Mg EDTA可以缓解阳离子猝灭，但阴离子的干扰可能需要在类似于土壤ATP提取物的化学成分的溶液中制备ATP测定标准。2001爱思唯尔科学有限公司版权所有。
Go-extracted ions with ATP from soils may interfere with ATP luciferin-luciferase luminescence reaction when ATP is assayed. The effects were investigated in a typical concentration range of cations and anions potentially extractable in soils. A commercial ATP assay product (Sigma Chemical Co.) was used. Significant quenching is evidenced from a concentration of 0.10 mM with Cu2+ and Zn2+, and 1.00 mM with Ca2+. The order of quenching at 1.00 mM was: Cu2+ > Zn2+ Ca2+ = Na+ = Mg2+, while Mg2+ = Mn2+, both Ca2+ and Na+ > Mn2+. The quenching was found to be much more severe with selected special heavy metal cations with quenching in the order: Ti3+ > Hg2+ > Cr3+. Because cation quenching can be alleviated by addition of EDTA, three forms of EDTA (Mg, Na and acid EDTA) were tested for their suitability for the assay. The Mg-EDTA was found superior to the other two. Presence of PO43- at concentrations of 0.01 and 0.05 mM, and NO3- at 0.01 and 0.10 mM, significantly enhanced ATP light emission (8-13%). However, SO42- at similar concentrations significantly decreased light emission. The quenching by CO32- and Cl- was only observed at high concentrations (3.20 mM and up). The order of quenching for the anions at a concentration of 6.4 mM was: PO43- > CO32- > SO42- > NO3- > Cl-. Enhanced or depressed light emission induced by ions would produce significant over or underestimation of soil ATP. While addition of Mg EDTA may alleviate cation quenching, the interference from anions may require the ATP assay standards be prepared in a solution of similar chemical composition to that in soil ATP extracts. (C) 2001 Elsevier Science Ltd. All rights reserved.