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Caged ATP - an internal calibration method for ATP bioluminescence assays
R M Calvert1, H C Hopkins, M J Reilly
1Department of Life Sciences, The Nottingham Trent University, Nottingham and Biotrace Ltd, Bridgend, UK.
Letters in Applied Microbiology
|April 4, 2000
Summary
Caged adenosine triphosphate (ATP) serves as an internal standard for ATP bioluminescence assays. Industrial cleansing solutions, ethanol, and pH significantly affect firefly luciferase activity, impacting hygienic monitoring in the food industry.
Area of Science:
- Biochemistry
- Analytical Chemistry
- Food Science
Background:
- Adenosine triphosphate (ATP) bioluminescence, utilizing the firefly luciferase system, is a key method for assessing hygiene in the food industry.
- Accurate hygienic monitoring requires understanding factors that may interfere with ATP detection.
Purpose of the Study:
- To evaluate caged ATP as a reliable internal standard for ATP bioluminescence.
- To quantify the impact of common industrial substances and conditions on firefly luciferase activity.
Main Methods:
- Utilized caged ATP as an internal standard in ATP bioluminescence assays.
- Quantified the effects of five industrial cleansing solutions, varying concentrations of ethanol, and a range of pH values on firefly luciferase activity.
Main Results:
- Cleansing solutions inhibited bioluminescence by 6-47% at working concentrations.
- 1% ethanol inhibited light output by 15%, while concentrations above 4% enhanced it.
- pH values below 4 or above 10 quenched the light signal by 20-25%.
Conclusions:
- Caged ATP is a viable internal standard for ATP bioluminescence assays.
- Industrial cleansing agents, ethanol concentration, and extreme pH levels can significantly interfere with hygienic monitoring results.
- These findings are crucial for optimizing ATP bioluminescence protocols in food industry hygiene assessments.