Preparation and evaluation of Orange Fruit Reference Extract based on the principles of Q-markers-A general procedure for establishing Herbal Reference Extracts used for identification

Abstract

Background Herbal Reference Extracts (HREs) have been used for the overall quality control (QC) of Traditional Chinese Medicines (TCMs) and used as an effective strategy for solving the shortage of reference standards in quality evaluation of TCMs in recently years. But the preparation and evaluation of it has rarely been reported. The ideal HREs should be a combination of Chemical Reference (CR) and Efficacy Reference (ER). However, it is not realistic to achieve "completely extracted" of TCMs at the present stage. Quality markers (Q-markers) provides a series of effective and scientific strategies for QC of TCMs. And Q-markers-based HREs should be a meaningful attempt to promote the standardization of quality evaluation system for TCMs. Purpose Preparation adequate HREs based on the principles of Q-markers, and used for quality evaluation of relevant preparation products. Then propose the general procedure for establishing HREs used for identification. Methods The authenticity, specificity, consistency and stability (ASCS) were the four essential principles of HREs. In this study, we chose Aurantii Fructus as an example, Firstly, selected 7 Q-markers of it which has the properties of specificity, effectiveness and testability based on the combination of analytic hierarchy process and entropy weight method (AHP-EWM). Secondly, preparated its reference extract which is called Orange Fruit Reference Extract (OFRE) based on ASCS and its Q-markers. Thirdly, established the QC method of OFRE according to its usage requirements. And finally, demonstrated its application of the thin layer chromatography (TLC) identification and characteristic chromatogram identification in the Orange fruit formula granule (or called Zhiqiao Peifangkeli, ZQPK). Results Compared with the Herbal Reference Substance (HRS) which with characteristic appearance or morphological structure, OFRE can be used for TLC identification of ZQPK conveniently, without complicated extraction or purification steps. And in the analysis of characteristic chromatogram of ZQPK, the OFRE can provid a rather simple profile, the main naringin, neohesperidin, nobiletin and tangeretin were clearly separated, which indicated that the OFRE could be used to identify the characteristic chromatogram of ZQPK and reduce the use of Chemical Reference Substance (CRS). And finally, we propose the general procedure for establishing HREs used for identification. Conclusion In summary, Q-markers-based HREs is feasibility, and should have a broader application prospect. And this research concept can serve as a reference for study on preparation, evaluation and application of HREs used for identification.