HPLC法考察60Co辐照对牛黄清感胶囊中4种有效成分的影响

Effects of 60Co Irradiation on Four Effective Components in Niuhuang Qinggan Capsules by HPLC

目的:考察60Co辐照灭菌对牛黄清感胶囊中4种有效成分含量的影响。方法:选择0,5,8,10 k Gy 4种辐照剂量对牛黄清感胶囊进行辐照,比较辐照前后需氧菌总数和霉菌、酵母菌总数的变化;采用HPLC法,Agilent ZORBAX SB-C18柱(4.6 mm×250 mm,5μm)色谱柱,乙腈-0.1%磷酸溶液为流动相,波长:210,447 nm;流速:1.0 ml·min^-1;柱温:30℃,进样量:10μl,比较有效成分绿原酸、黄芩苷、连翘苷、胆红素辐照前后含量的变化,并将所得数据用SPSS 22.0软件分析。结果:绿原酸、黄芩苷、连翘苷、胆红素线性范围分别为0.011~0.443μg·ml^-1(r=0.9995)、0.470~18.785μg·ml^-1(r=0.9997)、0.015~0.601μg·ml^-1(r=0.9989)、0.024~0.961μg·ml^-1(r=0.9991);平均加样回收率分别为100.79%、102.20%、99.43%、100.39%,RSD均小于2.0%(n=9)。牛黄清感胶囊经5 k Gy辐照剂量灭菌可达到中国药典2015年版微生物限度要求;经60Co辐照(0,5,8,10k Gy)后,牛黄清感胶囊中的绿原酸、黄芩苷、连翘苷含量辐照前后变化均无统计学意义(P>0.05);胆红素含量在辐照剂量8k Gy时辐照前后含量差异有统计学意义(P<0.05)。结论:低剂量60Co辐照对牛黄清感胶囊的灭菌效果显著,在辐照剂量不超过5 k Gy时,绿原酸、黄芩苷、连翘苷、胆红素的含量变化不显著,可为牛黄清感胶囊辐照灭菌手段提供参考。

Objective:To study the effects of 60Co irradiation on the contents of four constituents in Niuhuang Qinggan capsules.Methods:Niuhuang Qinggan capsules were irradiated under 0,5,8 and 10 k Gy of radiation doses,and then,the total aerobic plate account and the total yeasts&molds were compared before and after the irradiation.HPLC analysis was performed on an Agilent ZORBAX SB-C18 column(250 mm×4.6 mm,5μm)with the mobile phase consisting of acetonitrile-0.1%phosphoric acid solution.The detection wavelengths were set at 210 nm and 447 nm with the column temperature at 30℃.The flow rate was 1.0 ml·min^-1 and the injection volume was 10μl.The content changes of the active components including chlorogenic acid,baicalin,forsythin and bilirubin were compared before and after the irradiation,and the obtained data was analyzed by SPSS 22.0 software.Results:The linear range of chlorogenic acid,baicalin,forsythin and bilirubin was 0.011~0.443μg·ml^-1(r=0.9995)、0.470~18.785μg·ml^-1(r=0.9997)、0.015~0.601μg·ml^-1(r=0.9989)、0.024~0.961μg·ml^-1(r=0.9991),respectively.The average recovery was 100.79%,102.20%,99.43%and 100.39%,respectively,and the RSDs were all below 2.0%.Under the radiation dose of 5 k Gy,the microbial results of Niuhuang Qinggan capsules could reach the limit requirements of Chinese Pharmacopoeia;exposed to60Co irradiation at doses of 0,5,8 and 10 k Gy,chlorogenic acid,and forsythin were without statistically significant changes(P>0.05);after 8 k Gy radiation bilirubin had,statistically significant changes(P<0.05).Conclusion:The sterilization effect of60Co irradiation at low dose on Niuhuang Qinggan capsules is significant.When the irradiation dose is equal to or below 5 kGy,the content changes of chlorogenic acid,baicalin,forsythin and bilirubin are not obvious,which can provide reference for the radiation sterilization of Niuhuang Qinggan capsules.