| Objective:Hugan tablet、Bupleuri Radix and Isatidis Radix are commonly used Chinese traditional medicine.Their quality standards were recorded in Chinese Pharmacopoeia(version of 2015)with qualitive identification and assay of index component.However,there are still some deficiencies need to be improved,such as economic and simplicity.On the basis of their present quality standards,a systmetic research work was carried on to improve the quality control level,ensure the quality and clinical efficacy of drugs.This research work was partly supported by the research project "The standardization construction of Hugan tablets" from National TCM Standardization Project.Method:1.The HP LC fingerprints of Hugan tablet、Bupleuri Radix and Isatidis Radix were established.The chromatographic conditions were optimized,the fingerprint methodology was examined,and the similarity evaluation was performed.2.The HPLC-ESI-MS/MS was used to identify the main characteristic peaks to enrich the distribution of chemical constituents in the fingerprints.3.On the basis of the established fingerprints,The methods for content determination of Saikosaponin a,c,d in Bupleuri Radix;(R,S)-Goitrin,Adenosine in Isatidis Radix;(R,S)-Goitrin,Chlorogenic acid,Saikosaponin b2,Schisandrin,Deoxyschizandrin,Schisandrin B in Hugan tablet.The sample pretreatment method and method validation were examined.Result:The method validation results show that the established HPLC method is stable,accurate and has good separation effect for the detection of its fingerprints.The HPLC-ESI-MS/MS was used to identify the main characteristic peaks to enrich the distribution of chemical constituents in the fingerprints.There were 5 characteristic peaks of Bupleuri Radix.In addition to the great differences in the Bupleuri Radix medicinal materials from the S9 Inner Mongolian origin,the HPLC peaks of the other 19 batches of medicinal plants were consistent and the similarity was 0.844 to 0.997.There were 4 characteristic peaks of Isatidis Radix.In addition to the similarity of S17 Gansu Dingxi(Datong)was 0.762,the similarity of the remaining 19 batches was 0.865-0.989.There were 11 characteristic peaks of the Hugan tablets.The similarity of 15 batches of the same manufacturer(Heilongjiang Kuihua Pharmaceutical)and the 15 batches of different manufacturers Hugan tablets was higher than 0.90.Based on the established fingerprints,the methods for content determination of multi-indicator components was established.Validation results showed those methods were precise,stable,reproducibility,and accurate for the quality control.The results of the determination of the contents indicate that the content of the index components in different manufacturers or in different production areas is significantly different,which may be related to factors such as environment and processing place.Conclution:In this study,fingerprints were combined with multi-component content determination to achieve the unity of fuzzy characterization and quantitative control,improving the efficiency of testing;The use of high performance liquid chromatography mass spectrometry to identify the main characteristic peaks in the fingerprints completes the distribution of chemical constituents in the fingerprints;The conclusion is more comprehensive and accurate than a single measurement and provides a basis for the overall quality evaluation and control of Hugan tablets,Bupleuri Radix and Isatidis Radix. |
