刺五加注射液中过敏性杂质的分析

Analysis of Allergic Dopants in Ciwujia Injection

目的探索刺五加注射液中的过敏性杂质,建立相应的检查方法。方法针对刺五加注射液"水煮醇沉"工艺的特点提取刺五加抗原,采用豚鼠被动皮肤免疫(PCA)实验寻找刺五加注射液中的过敏性杂质,用间接ELISA法检测过敏性杂质。结果PCA实验证实,ELISA法阳性刺五加注射液可以引发刺五加抗原致敏豚鼠的速发型过敏反应;采用间接ELISA法检测刺五加注射液中的过敏性杂质,其灵敏度为0.5 mg·L^(-1),线性范围为4～128 mg·L^(-1)(r^2=0.9988),平均加样回收率为86.3%;对92批市场流通样品进行了测定,8批(8.7%)检出过敏性杂质;经超滤处理后,样品的ELISA实验呈阴性,而其UFLC分析显示,超滤前后药物中的HPLC图谱没有变化。结论本实验结果表明,药材中的植物蛋白等过敏性杂质可能会残留到注射液中导致过敏反应的发生,间接ELISA法可用于测定刺五加注射液中的过敏性杂质;超滤技术可以去除刺五加注射液中的过敏性杂质。

OBJECTIVE To screen allergic dopants and establish an ELISA method for the determination of allergic dopants determination method in Ciwujia injection. METHODS By the method of water boiling and precipitation with ethanol, acanthopanax senticosus antigen were extracted from acanthopanax senticosus harms. The allergic dopants contained in the Ciwujia injection were confirmed by the passive cutaneous anaphylaxis（PCA） test and were further determined by the indirect ELISA method. RESULTS Both of the ELISA positive products and acanthopanax senticosus antigen extracted from the sacanthopanax senticosus harms induced the immediate hypersensitivity reaction;The calibration curves of ELISA used to determined the allergic dopants was linear at the concentration range of 4 - 128 mg·L^-1 （r^2 = 0. 998 8） and the average recovery of acanthopanax senticosus antigen were 86. 3%. Eight （8.7%） batch of samples were positive in total 92 bacthes of Ciwujia injections. After being treated by the ultra-filtering method, ELISA positve samples became negative while there were no changes in the content of principal composition in Ciwujia injection. CONCLUSION The allergic dopants such as residual vegetable protein from raw material might induce allergic response; the indirect ELISA established in our research could determine the concentration of allergic dopants. The ultra-filtering method could remove the allergic dopants from Ciwujia injection.