Objective To reduce experimental costs and improve the utilization of S9, we use spiral coating technique to evaluate the activity of rat liver S9 prepared by combination inducing method as well as to establish cryopreservation method.Methods Using spiral coating technique and Ames test to evaluate the activity of self-made rat liver S9 and commercially available S9 separately.We use glycerinum as protective agent to establish cryogenic storage method, so that S9 can be in liquid form stored at -20 °C in the refrigerator.Results In the Ames assay as well as using spiral coating technique, the number of revertant colonies had dose-response relationship among the dose of S9.When conditions were the same, the number of revertant colonies in positive control was at the approximate level in presense of self-made rat liver S9 and commercially available S9 respectively.When S9 ( concentration of 38%) was added to the amount at 1.48 ~6.62 μL /μL broth dose of bacteria, it can significantly induced Salmonella typhimurium histidine strains TA100, reverse mutation rates were three times more than the control group.Conclusions Spiral coating technique can successfully evaluate the activity of rat liver S9.The inducing method of combination of PB and BF can take the place of the unducing method of PCBs in the preparation of Liver homogenate S9.%目的：建立螺旋涂布法，评价本实验室用联合诱导法制备大鼠肝匀浆S9活性优劣，以期大大降低实验成本，并用Ames实验法评价所建立的S9低温保存方法的可行性，提高S9利用率。方法采用螺旋涂布法和Ames试验方法对市售和自制两种来源S9活化的阳性结果进行比较，并加入甘油保护剂，建立S9酶系统低温储存方法，使其以液态保存于－20℃冰箱中。结果 Ames实验中市售和自制两种S9在阳性组中都显示出明显活化作用，同一条件下，Ames试验中使用两种S9所得阳性结果并无较大差异，变率间差异无统计学意义。螺旋涂布法中，同一条件下，使用两种S9所得阳性结果也无较大差异，突变率间差异无统计学意义， S9（浓度为38％）加入量在1.48～6.62μL／μL菌液剂量下，对TA100菌有明显诱导作用，回复突变率均为对照组的3倍以上。结论可以用螺旋涂布法成功评价大鼠肝S9活性大小，联合诱导法可以替代多氯联苯诱导法制备肝匀浆S9。
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