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ATP-bioluminescence technique for fruit quality assessments: a detailed methodology for analysis of adenylate nucleotides

The bioluminescence technique has been used during a long time for measurementof adenylate nucleotides (ATP, ADP, and AMP) in several investigation fields.Until the end of the seventies, the assay was complicated and the results uncertainbecause of the low purity reagents, which were initially, extracted directly fromdesiccated lanterns of firefly (Photinus pyralis), and poorly controlled reactionconditions at the laboratory. With the introduction of a new type of luciferinluciferasereagent, the ATP Monitoring Reagent (AMR), these disadvantages havebeen overcome, and therefore, the application of the assay has been extended.The new reagent results in the emission of light of an almost constant intensity,proportional to the ATP concentration. This is in contrast to the flash obtainedwith conventional reagents. This new technique facilitates the measurements andimproves the sensitivity of the assay. The reagent may also be used to monitorchanges in ATP concentration because the ratio between the ATP concentration andlight intensity remains constant. This work describes the principle and a detailedmethodology for determination of ATP, ADP, and AMP by the luciferin-luciferasereaction in fruits such as apples, pears and sweet cherries and some applications.