中药跌打膏质量标准提升与安全性研究

Quality Standard Improvement and Safety Study of Chinese Medicine Dieda Ointmen(t 中药跌打膏)

目的 建立中药跌打膏质量控制方法,验证中药跌打膏安全性,为临床提供参考。方法 根据配方组成,采用薄层色谱法(TLC)对方中川乌、草乌、大黄、续断、泽兰进行定性鉴别,采用高效液相色谱法(HPLC)定量测定中药跌打膏中大黄素,以用于对中药跌打膏的质量控制;此外,通过皮肤急性毒性实验、皮肤刺激性实验、皮肤过敏性实验以及豚鼠皮肤组织病理学检查对中药跌打膏的安全性进行评价。结果 该实验中川乌、草乌、大黄、续断、泽兰定性鉴别方法操作简便,分离度好,专属性强。大黄素平均加样回收率为84.917 3%,RSD为1.466 1%。大黄素含量范围为1.099 9%~1.123 8%;精密度、稳定性、重复性试验的RSD分别为0.229 4%、1.835 9%、1.025 0%,均小于3.00%(n=6);安全性实验中,中药跌打膏对小鼠皮肤无刺激性,对小鼠体质量、肝肾功能无影响。一次给药后,14 d观察期间各组小鼠行为活动、精神状态、饮食情况及排便均未出现异常;给药14 d后,取小鼠血液进行血常规检查,后对小鼠进行解剖取血,血常规各项指标及肝肾功能无异常,各脏器未出现肉眼可见病理变化;中药跌打膏干预后的豚鼠皮肤病理检查均未见组织病理学改变。中药跌打膏对豚鼠皮肤无刺激性,中药跌打膏对豚鼠皮肤无明显致敏。结论该实验建立的中药跌打膏中川乌、草乌、大黄、续断、泽兰的TLC鉴别方法和大黄素含量测定方法有良好的精密度、稳定性和重复性,可用于中药跌打膏的质量控制。中药跌打膏对小鼠局部皮肤无毒性,对豚鼠皮肤无刺激性,对豚鼠皮肤无致敏性,说明中药跌打膏外用治疗疾病安全可靠。

Objective To establish a quality control method for Chinese Medicine Dieda Ointment(中药跌打膏),to verify the safety of Chinese Medicine Dieda Ointment and to provide clinical reference.Methods Based on the composition of the formula,the qualitative identification of Chuanwu(Aconiti Radix),Caowu(Aconiti Kusnezoffii Radix),Dahuang(Rhei Radix Et Rhizoma),Xuduan(Dipsaci Radix)and Zelan(Lycopi Herba)were performed by thin-layer chromatography(TLC),and the quantitative determination of rhodopsin in Chinese Medicine Dieda Ointment by high performance liquid chromatography(HPLC),which was used for the quality control of Chinese Medicine Dieda Ointment;the safety of Chinese Medicine Dieda Ointment was evaluated by skin acute toxicity test,skin irritation test,skin allergy test and guinea pig skin histopathological examination.The safety of the cream was evaluated by skin acute toxicity test,skin irritation test,skin allergy test and guinea pig skin histopathology.Results The qualitative identification method of Chuanwu(Aconiti Radix),Caowu(Aconiti Kusnezoffii Radix),Dahuang(Rhei Radix Et Rhizoma),Xuduan(Dipsaci Radix)and Zelan(Lycopi Herba)were simple,good separation and specificity.The average recovery of rhodopsin was 84.9173%with the RSD of 1.4661%.The range of rhodopsin content was 1.0999%-1.1238%;the RSDs of precision,stability and reproducibility tests were 0.2294%,1.8359%and 1.0250%,which were less than 3.00%(n=6);In the safety experiment,the Chinese Medicine Dieda Ointment had no irritation to the skin of mice,and no effect on the body mass,liver and kidney function of mice.After one administration,no abnormalities were found in behavioral activities,mental status,diet and defecation of mice in each group during 14 d observation period;after 14 d of administration,blood was taken from mice for routine blood examination,and later mice were dissected and blood was taken,and there were no abnormalities in various indicators of blood routine and liver and kidney functions,and no pathological changes were seen in the organs;no histopathological changes were seen in the skin pathological examination of guinea pigs after the intervention of Traditional Chinese Medicine Dieda Ointment.No histopathological changes were observed in the skin pathological examination of guinea pigs after the intervention of Chinese Medicine Dieda Ointment.There was no irritation of the skin of guinea pigs by the herbal Dieda Ointment,and no significant sensitization of the skin of guinea pigs by the herbal Dieda Ointment.Conclusion The TLC method for the identification of Chuanwu(Aconiti Radix),Caowu(Aconiti Kusnezoffii Radix),Dahuang(Rhei Radix Et Rhizoma),Xuduan(Dipsaci Radix)and Zelan(Lycopi Herba)in Chinese Medicine Dieda Ointment and the method for the determination of rhodopsin content established in this experiment have good precision,stability and reproducibility;they can be used for the quality control of Chinese Medicine Dieda Ointment.Chinese Medicine Dieda Ointment was not toxic to the local skin of mice,not irritating to the skin of guinea pigs,and not allergenic to the skin of guinea pigs,indicating that Chinese Medicine Dieda Ointment is safe and reliable for topical treatment of diseases.