Study On ATP Extractor In ATP-Bioluminescence

ATP-Bioluminescence method allowed the rapid quantification of microorganisms by measuring their intracellular ATP in the firefly luciferin-luciferase BL reaction. The biochemistry of the bioluminescent reaction was first described in the late 1940s, and its use in the determination of ATP content using the luciferin-luciferase system was first described in 1949. Since then it developed to a very accurate means of measuring levels of ATP. HACCP was internationally recognized, which demanded higher standards of method to detect microorgansim. It was necessary to choose an ATP extractor with high efficiency to improve the BL sensitivity.This paper concerned with several factors on ATP extractor including: the selection of an ATP extractor and ATP extracting measurement; the optimization of factors for ATP extraction; the preliminary study on the mechanism of BAB on bacteria; the comparison of ATP-Bioluminescence measurement and plate colony counting in detecting the microorganisms in the air, cooked beef and milk.The concentration of and the extracting time of the quaternary ammonium salt cation surfactant as ATP extractor were established; some important compounding factors for improving the BL intensity were also studied and optimized such as heating, microwave, acid and alkali solution, Cu²⁺, PHMG and the ingredients in the buffer solution.The bacterial cells were observed under the transmission electron microscope, and the leakage of bacteria was detected by ultraviolet ray at 260nm after bacteria exposuring to BAB. The results indicated that the mechanism of effect of BAB on E.coli and S.aureus was mainly destruction of permeability barrier of the bacterial cells. The comparison of BAB and the two other saled ATP extractors by HPLC showed that the quaternary ammonium salt cation surfactant was creative as ATP extractor.The process to determine the bacteria in environmental air, meat and milk rapidly using ATP-Bioluminescence method was investigated and compared with traditional plate colony counting method. The results showed that there was a significant correlation between these two methods.