八珍益智合剂3种成分的含量测定及稳定性考察

Investigation on Contents and Stability of Three Components of Bazhen Yizhi Mixture

关键词:  八珍益智合剂  一测多评  甘草苷  橙皮苷  芹糖甘草苷  稳定性

Key words:  Bazhen Yizhi mixture  QAMS  liquiritin  hesperidin  liquiritin apioside  stability

摘要:

目的 建立一测多评法同时测定八珍益智合剂中甘草苷、芹糖甘草苷和橙皮苷3种成分的含量，并对八珍益智合剂3种成分的稳定性进行考察。方法 采用HPLC-DAD法，以橙皮苷为参照物，测定其他2种成分的相对校正因子和相对保留时间，并计算各成分含量；以外标法为对照，比较一测多评法（single marker，QAMS）与外标法（external standard method，ESM）实测值的差异，探讨一测多评法的可行性。同时测定八珍益智合剂在高温、强光照射、加速试验和长期室温条件下的稳定性，为其贮藏条件和保质期提供数据。结果 经过方法学验证，3种成分在4.512~108.2，10.13~101.30，4.496~107.90 μg·mL^-1内线性关系良好（r>0.999）；平均加样回收率为95.9%~104.7%，RSD ≤ 3%；2种成分相对橙皮苷的相对校正因子分别为0.49和0.38，且在不同实验条件下相对校正因子重复性良好；含量测定QAMS计算结果与ESM实测值无明显差异。稳定性考察结果表明，在高温条件下长时间放置芹糖甘草苷和橙皮苷含量下降速度较快，甘草苷含量下降较为平缓，提示中药合剂在高温条件下不宜放置过长时间。结论 本研究所建方法准确可靠、重复性好，可用于八珍益智合剂的质量控制，本合剂应密封置阴凉处保存。

Abstract:

OBJECTIVE To investigate the stability of Bazhen Yizhi mixture and to develop a quantitative analysis of simultaneous determination of liquiritin, liquiritin apioside and hesperidin by single-marker method. METHODS The analysis of this mixture was performed on HPLC-DAD and by using hesperidin as a standard reference, the relative correction factors(RCFs) and relative retention time of the other two constituents were measured and calculated. Moreover, comparison of the difference between the quantitative analysis of multicomponents by single marker(QAMS) and external standard method(ESM) was used to explore the feasibility of QAMS. Meanwhile, the stability of Bazhen Yizhi mixture was investigated under high temperature, bright lights, acceleration and long-term room temperature test conditions for its storage conditions and expiration dating to provide data. RESULTS By methodological study, three constituents showed good linear relationships within 4.512-108.2, 10.13-101.30, 4.496-107.90 μg·mL^-1(r>0.999), whose average recoveries were 95.9%-104.7% with the RSDs of ≤ 3%. The RCFs of liquiritin apioside and liquiritin were calculated by methods validated as 0.49 and 0.38, respectively. The determination results reflected the good reproducibility of the RCFs in different influence factors. There was no significant difference between the calculated values by QAMS method and the measured values by ESM. The results of stability investigation showed that the contents of liquiritin apioside and hesperidin decreased rapidly under high temperature condition for long time, while liquiritin decreased gently, implied that the storage time of traditional Chinese medicine mixture should not be too long. CONCLUSION The method applied in this study is accurate, reliable, reproducible, and can be used for the quality control of Bazhen Yizhi mixture. This mixture should be sealed and stored in a cool place.