Colorimetric method for enzymatic screening assay of ATP using Fe(III)-xylenol orange complex formation

Analytical and Bioanalytical Chemistry
Akihiko IshidaTamio Kamidate

Abstract

In hygiene management, recently there has been a significant need for screening methods for microbial contamination by visual observation or with commonly used colorimetric apparatus. The amount of adenosine triphosphate (ATP) can serve as the index of a microorganism. This paper describes the development of a colorimetric method for the assay of ATP, using enzymatic cycling and Fe(III)-xylenol orange (XO) complex formation. The color characteristics of the Fe(III)-XO complexes, which show a distinct color change from yellow to purple, assist the visual observation in screening work. In this method, a trace amount of ATP was converted to pyruvate, which was further amplified exponentially with coupled enzymatic reactions. Eventually, pyruvate was converted to the Fe(III)-XO complexes through pyruvate oxidase reaction and Fe(II) oxidation. As the assay result, yellow or purple color was observed: A yellow color indicates that the ATP concentration is lower than the criterion of the test, and a purple color indicates that the ATP concentration is higher than the criterion. The method was applied to the assay of ATP extracted from Escherichia coli cells added to cow milk.

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Citations

May 21, 2013·Enzyme and Microbial Technology·Victor M BalcãoMarta M D C Vila
Jan 7, 2019·Analytical and Bioanalytical Chemistry·Sathya SrinivasanBhaskar Mohan Murari
Feb 28, 2021·Environmental Science and Pollution Research International·Sindhunath Chakraborty, Arun Kumar Mishra

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