红细胞溶血试验作为眼刺激性替代方法的初步研究

Preliminary study on red blood cell haemolysis assay as an alternative method for eye irritation test

目的探讨红细胞溶血试验在化妆品原料和产品眼刺激性检测中的适用性。方法选择已知动物实验眼刺激性分级的23种化学品原料和12个洗发、护发类化妆品产品作为受试物,确定受试物引起绵羊红细胞50%发生溶血时的受试物浓度(LC50值)和血红蛋白变性指数(DI,%),计算L/D值(LC50/DI),以L/D值作为评价标准,进行眼刺激性分级。将红细胞溶血试验刺激性分级结果与动物实验结果进行分级一致性比较,初步分析体外试验的适用性。结果红细胞溶血试验对23种化妆品原料的分级结果与动物实验的分级一致性系数为0.582,而当化妆品原料不包括酸、碱及难溶于水的物质时,一致性系数为0.803。红细胞溶血试验对化妆品的分级结果与动物实验结果具有等级相关性(γ=1.000,P<0.001)及分级一致性(κ=0.733,P<0.001)。结论与Draize试验比较,红细胞溶血试验具有较好的预测能力,除了酸类、碱类以及难溶性受试物,作为严重眼刺激性化妆品原料和产品评价的筛选方法具有良好的应用价值。

Objective To study the possibility of red blood cell（RBC） haemolysis assay as an alternative method for eye irritation test of cosmetic materials and products.Methods Twenty-three cosmetic materials and 12 cosmetic products were evaluated with RBC haemolysis assay and eye irritation test in the study.The median hemolytic concentration（LC50）and the denaturation index（DI）were calculated,and the LC50/DI ratio was used for determination of potential classification of eye irritation.The result of in vivo test was compared statistically with that of in vitro experiment.Results The rank correlation coefficients were 0.582 and 0.803 between the results of haemolysis assay and Draize test,respectively,with and without acids,alkalies and insoluble materials.The rank concordance coefficient was 0.733 between the results of in vitro and in vivo tests.Conclusion The results suggested that RBC haemolysis assay could be used as a screening test of eye irritation evaluation for cosmetic products without containing acids,alkalies and insoluble materials.