Research On Quality Standards Of Curcuna Pieces Based On QAMS And Fingerprint

Curcumae Rhizoma is derived from rhizomes of three species of Curcuma L.,which belongs to Zingiberaceae,including Curcuma phaeocaulis,Curcuma kwangsiensis and Curcuma wenyujin.It was reported to have the effect of traveling qi and breaking plasma and elimination produit and analgesia with acrid and bitter flavor.It converges to liver and spleen channel.Curcumae Rhizoma was reported to possess wide activities of anti-liver fibrosis,liver protect,anti-tumor and anti-thrombotic,which has great development potential and broad market prospects.The promoting blood circulation and removing blood stasis effect of Curcumae Rhizoma was enhanced and it mainly converges to liver and spleen channel after vinger-processed.Sesquiterpenes and curcuminoids are thought to be the main two categories of biological active ingredients.determination of Curcuma herbs and Pieces in Pharmacopoeia was that the content of essential oil in Herbs not less than 1.5%(ml/g),and 1.0%(ml/g)for Pieces.It dosen’t determine the indicative active ingredient for the quantitative analysis.And it only reproduces crude Pieces and vinegar-boiled Pieces,not included vinegar-biked Pieces that commonly used in clinical.The direction of development of "From single to multi-index index from the index components to control the medicinal ingredient" has been established with 2010 edition of "Chinese Pharmacopoeia" as the representative of the quality standard.In response to these problems,this paper studied sesquiterpenes and curcuminoids in crude Pieces,vinegar-biked Pieces and vinegar-boiled Pieces with QAMS and fingerprint techniques.This paper studied the specificity in fingerprints to develop a more comprehensive system of quality assessment method of Curcuma Pieces.QAMS Research of Curcuma Pieces:In this study,a quantitative analysis of multi-components by single marker(QAMS)method for simultaneous determination of sesquiterpenes and curcumin components in different processed Curcuma Rhizomae has been developed.The Chromatographic conditions was as following:A Hanbon ODS-2 C18 column(250 mm×4.6 mm,5μm)was used for chromatographic separation at a column temperature of 30℃.The mobile phase was a mixture of acetonitrile containing 0.05%formic acid(mobile phase A)and water containing 0.05%formic acid(mobile phase B)in eluted gradient.The flow-rate was 1.0 ml/min and the injection volume was 20 μl.The analytes were monitored with DAD at 216 and 430 nm.All the components showed good linearity and had a wide linear content range.The RSD of precision was less than 2.66%,RSD of stability was less than 2.76%,RSD of repeatability was less than 2.64%.Their average recoveries were 101.84%、98.67%、100.47%、101.32%、100.12%、97.59%、103.73%and 102.64%,and the RSD was less than 2.75%.The easily available and stable active component germacrone was selected as the internal reference substance for calculating the relative correction factor(RCF)of the other sesquiterpenes and curcuminoids,including curdione,curcumol,furanodiene,P-elemene,bisdemethoxycurcumin,demethoxycurcumin and curcumin.Different reversed-phase columns and different instruments,were used to investigate the variation of the RCFs.The RCFs had good reproducibility(RSD<2.47%)for different columns and instruments,demonstrating that the RRFs could be used for HPLC quantitative analysis.The contents of 8 components in different processed Curcumae Rhizoma samples were determined by external standard method and QAMS,respectively.The relative error method,Pearson coefficient,correlation coefficient and the cosine of the angle were used to compare the results determined by external standard method and QAMS.The results came to be that the relative error were less than 3.0%,Pearson coefficient,correlation coefficient and the cosine of the angle were greater than 0.995.It could be seen that no significant differences were found in the quantitative results of 5 sesquiterpenes and 3 curcuminoids in different processed Curcuma Rhizomas by RCFs and external standard method.Fingerprint Research of Curcuma Pieces:This paper collected and prepared Curcuma Pieces of different origin,different varieties and different processing methods of 10 batches each.The method of sample preparation(including solvent extraction,extraction method,extraction time)and chromatographic conditions(mobile phase,column,gradient elution,column,detection wavelength)were investigated and optimized.The fingerprint of Curcuma Pieces of different species and different processing methods were established.Their similarity was evaluated by the software of State Pharmacopoeia Commission recommended "chromatographic fingerprint evaluation system".Results showed that the similarity of Curcuma Pieces of the same species and sameprocessing methods was greater than 0.9,which was comply with the requirements.This paper cluster analyzed Curcuma Pieces of different species and different processing methods.The results came out to be that Curcuma Pieces of the same species and same processing methods can be classified as a class,while that of different species and different processing methods can be classified as a class because of the low degree of similarity.Quality Standards Research of Curcuma Pieces:According to the relevant provisions in 2010 edition of "Chinese Pharmacopoeia",this paper studied Curcuma Pieces of different origin,different varieties and different processing methods from traits,identification,inspection and determination.The results showed that:the relevant provisions of traits,identification and inspection items of Curcuma Pieces in the current pharmacopoeia standards were feasible.However,determination should be improved.This paper added the species of vinegar-biked Curcuma Pieces,the TLC qualitative identification of the 4 active ingredient,and established the determination of three kinds of terpene active ingredients(curdione,curcumol and germacrone)in Curcuma Pieces based on preliminary studies in order to further improvingthe quality control system of Curcuma.The three active ingredients were relatively high and stable nature of the content,which Curcuma Pieces of different origin and different processing methods have in common.In summary,this paper developed a quality specifically evaluation method of crude,vinegar-biked and vinegar-boiled Curcuma Pieces through the research of QAMS,quality standards and fingerprint on Curcuma Pieces.This provides a scientific theory in quality monitoring Curcuma Pieces.