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目的:检测双黄连注射剂中1,3-二咖啡酰奎宁酸致敏家兔后血清中特异性抗体效价。方法:双黄连注射剂中疑似小分子半抗原物质1,3-二咖啡酰奎宁酸与正常家兔血清体外孵育制备完全抗原,免疫家兔获得特异性抗体。用ELISA法检测抗血清的抗体效价。间接竞争ELISA法测定血清特异性并绘制抑制曲线。采用兔免疫球蛋白E(Ig E)ELISA试剂盒检测各组家兔抗血清中IgE抗体的含量。结果:1,3-二咖啡酰奎宁酸的抗体效价(A)值高于阴性对照组2倍,说明其具有潜在的致敏性。回归方程I=0.170 6lg C+0.317 5,相关系数r=0.985 4,检测限IC10为57.40μg·ml~(-1),半数抑制浓度IC50=8.732 0mg·ml~(-1)。家兔体内明显产生了外源性IgE型抗体。结论:在本实验条件下双黄连注射剂中疑似半抗原物质1,3-二咖啡酰奎宁酸具有致敏性。
OBJECTIVE: To detect the serum specific antibody titer of Shuanghuanglian injection in rabbits sensitized with 1,3-dicaffeoylquinic acid. METHODS: Shuanghuanglian injection was used to inoculate the suspected small molecule haptens, 1,3-dicaffeoylquinic acid, with normal rabbit serum to prepare the complete antigen and immunized rabbits to obtain specific antibodies. Antiserum antibody titers were measured by ELISA. Indirect competitive ELISA was used to measure the serum specificity and draw the inhibition curve. Rabbit immunoglobulin E (Ig E) ELISA kit was used to detect the content of IgE antibody in rabbit antiserum. Results: The antibody titer (A) of 1,3-dicaffeoylquinic acid was 2 times higher than that of the negative control group, indicating that it had potential allergenicity. The regression equation I = 0.170 6lg C + 0.317 5, the correlation coefficient r = 0.985 4, the limit of detection IC10 was 57.40μg · ml -1, the median inhibitory concentration (IC50) was 8.732 0mg · ml -1. Rabbit obviously produced exogenous IgE antibodies. CONCLUSION: 1,3-D-caffeoylquinic acid, a suspected hapten, in Shuanghuanglian injection is sensitized under this experimental condition.