Microplate Luminometry for Toxicity Bioassay of Chemicals on Luciferase
GE Hui-lin1, 2，LIU Shu-shen2*，CHEN Fu2，LUO Jin-hui1，Lü Dai-zhu1，SU Bing-xia1
1. Hainan Key Laboratory of Tropical Fruit and Vegetable Products Quality and Safety，Analysis and Testing Center，Chinese Academy of Tropical Agricultural Sciences，Haikou 571101，China 2. Key Laboratory of Yangtze Aquatic Environment，Ministry of Education，College of Environmental Science and Engineering，Tongji University，Shanghai 200092，China
Abstract：A new microplate luminometry for the toxicity bioassay of chemicals on firefly luciferase, was developed using the multifunctional microplate reader (SpectraMax M5) to measure the luminous intensity of luciferase. Efects of luciferase concentration, luciferin concentration, ATP concentration, pH, temperature, and reaction time on the luminescence were systematically investigated. It was found that ATP exerted a biphasic response on the luciferase luminescence and the maximum relative light units (RLU) occurred at an ATP concentration of 1.1×10-4 mol·L-1. The method was successfully employed in the toxic effect test of NaF, NaCl, KBr and NaBF4 on luciferase. Using nonlinear least square technique, the dose-response curves (DRC) of the 4 chemicals were accurately fitted with the coefficient of determination (R2) between the fitted and observed responses being greater than 0.99. The median effective concentration (EC50) of the 4 chemicals were accurately measured from the DRC models. Compared with some literatures, the bioassay is a fast easy-operate and cost-effective method with high accuracy.
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