蛋白核小球藻(Chlorella pyrenoidosa)微板毒性分析方法优化

以蛋白核小球藻(Chlorella pyrenoidosa，CP)为指示生物，96孔微板为暴露载体，污染物对藻的72 h生长抑制率为毒性指标，通过系统地检测蛋白核小球藻的生长曲线和吸收光谱，确定藻细胞密度和683 nm波长处光密度(D₆₈₃)之间的线性关系，考察不同初始藻密度、照度、暴露时间和暴露体积对藻生长的影响，建立了蛋白核小球藻微板毒性分析方法(CP-MTA法).将CP-MTA法应用于重金属盐、除草剂、杀虫剂以及离子液体等8种化学品对蛋白核小球藻的生长抑制毒性测试，以pEC₅₀为毒性指标，毒性大小顺序为敌草快>CuSO₄·5H₂O≈CdCl₂·2.5H₂O>氯化1-甲基-3-辛基咪唑(［Omim］Cl)>草甘膦>氯化1-甲基-3-丁基咪唑(［Bmim］Cl)>敌敌畏>乐果，与文献结果一致.CP-MTA法由于以微板为反应载体，所需样品少，便于多次平行，数据重复性好. 

Optimization of Microplate Toxicity Analysis Method Based on Chlorella Pyrenoidose

Taking Chlorella pyrenoidosa(CP) as an indicator organism,a 96-well microplate as an exposure carrier,and 72 h algae growth inhibition rate as a toxicity endpoint,a microplate toxicity analysis based on CP(CP-MTA) was developed.The growth curve and absorption spectra of CP were detected systematically.The linear relationship between the algal cell density and optical density at a wavelength of 683 nm was determined.The effects of initial cell density,illumination,exposure time,and exposure volume on the algae growth were examined.CP-MTA was employed to determine the growth inhibition toxicities of eight chemicals including heavy metal salts,herbicides,pesticides and ionic liquid on CP.It was found that if the pEC50 value was taken as a toxicity biomarker,the order of toxicity was:diquat > CuSO4·5H2O ≈ CdCl2·2.5H2O > 1-methyl-3-octyl-imidazolium chloride > glyphosate > 1-ethyl-3-methyl-imidazolium chloride > dichlorvos > dimethoate,which is consistent with other results in the literature.CP-MTA has some advantages such as small sample volumes,convenience for parallel testing,and good repeatability due to taking a microplate as the exposure carrier.