Quality Evaluation Of Dispensing Granules Made Of Yinchenhao Tang Based On Standard Decoction

Objective:To evaluate the quality of dispensing granules of Artemisiae co paSria Herba(ASH),Gardeniae Fructus(GF),and Rhei et Rhizoma(RRR),which made of Yinchenhao Tang.Methods:To prepare 15 batches of standard decoction of ASH,23 batches of standard decoction of GF and RRR.To determine the HPLC fingerprints of standard decoction and its raw materials,demarcate the common peaks and calculate the area of the common peaks.The common peaks were identified by Q-TOF/MS technology and the components confirmed by comparison of the control samples,which were included the quantity transfer rule of the standard decoction that was analyzed by the common peak transfer,the ratio of quantitative peak area of common peak,the index component transfer rate and the extract rate.Gathering dispensing granules from different manufacturers,which includes 28 batches of ASH,21 batches of GF,21 batches of RRR.Setting-out HPLC fingerprint and establishing a control fingerprint,calibrating common peak and making a calcultion of common peak area.Using Q-TOF/MS technology to identify the common peaks and determine content of the ingredients in the formula granules corresponding to the respective standard decoction.To go on the comprehensive analysis between formula granules and standard decoction on the common peak number,peak area ratio and index component ratio.At the same time,cluster analysis,principal component analysis,factor analysis and other pattern recognition were also be usedResults:(1)According to HPLC fingerprint,there were 17 peaks on ASH decoction pieces and 16 peaks on ASH standard decoction.The transfer rate was 94.12%.13 components in the common peak were confirmed by comparision of the reference substance,which were 1-caffeoylquinic acid,neochlorogenic acid,chlorogenic acid,caffeic acid,cryptochlorogenic acid,p-hydroxyacetophenone,1,3-dicaffeoylquinic acid,rutin,hyperoside,isoquercetin,3,4-dicaffeoylquinic acid,3,5-dicaffeoy lquinic acid and 3,4-dicaffeoylquinic acid.Caffeinylquinic acid glucoside and flavone diglycoside apigenin-6,8-C-diglucoside were detected for the first time in ASH.The ratio of the common peak area of standard decoction and decoction pieces showed a good positive linear relationship with the component transfer rate.The average transfer rate of neochlorogenic acid,cryptochlorogenic acid,1,3-dicaffeoylquinic acid and 3,4-dicaffeoylquinic acid exceeded 100%.The average transfer rate of chlorogenic acid and 3,5-dicaffeoylquinic acid were only 49.31%and 25.13%.Which suggested that the organic acids may be transformed into each other,chlorogenic acid may be partially transformed into cryptochlorogenic acid and neochlorogenic acid,3,5-dicaffeoylquinic acid may be partially transformed into l,3-dicaffeoylquinic acid and 3,4-dicaffeoylquinic acid.Rutin and hyperoside were 31.36%and 28.36%,respectively.The average transfer rates of other components were 50%～70%,and the average paste yield of standard decoction was 19.76%.14 peaks were transmitted from ASH Standard decoction to dispensing granules,the average content of components in the dispensing granules was significantly different from that in the standard decoction.The average content of chlorogenic acid was 41.67%of that in the standard decoction,and the average content of 3,5-dicaffeoylquinic acid was 43.11%,but the contents of neochlorogenic acid,cryptochlorogenic acid,1,3-dicaffeoylquinic acid and 3,4-dicaffeoylquinic acid were 139.44%,98.33%,119.04%and 78.88%.It was suggested that the related components were further transformed in the preparation process of formula particles due to heating and drying.Cluster analysis showed that ASH dispensing granules and standard decoction could be grouped into two groups,and dispensing granules produced by different enterprises could also be grouped together.The results of principal component analysis and factor molecular comprehensive score showed that the quality of ASH dispensing granules was significantly inferior to that of standard decoction.(2)According to HPLC fingerprint,there were 26 peaks on GF decoction pieces and 24 peaks on GF standard decoction.The transfer rate was 92.31%.15 components in the common peak were confirmed by comparision of the reference substance,which were shanzhiside,geniposidic acid,deacetyl asperulosidic acid methyl ester,hydroxygardenoside,ixoside,scandoside methyl ester,Jasminoside B,chlorogenic acid,gentiobioside,geniposide,rutin,isoquercitrin,crocin Ⅰ,crocin Ⅱ and quercetin.By comparing the mass spectrum data with the literature and the data of the compound library,nine new compounds were inferred.The ratio of the common peak area of standard decoction and decoction pieces showed a good positive linear relationship with the component transfer rate.The transfer rate of components in GF standard decoction is higher,which may be the components in GF were water-soluble.The average transfer rate of deacetyl asperulosidic acid methyl ester and scandoside methyl ester were exceeded 100%,the average transfer rate of shanzhiside was close to 100%,the average transfer rate of geniposide was only 80.43%,the average transfer rate of crocin I and crocin II were 48.48%and 40.64%.It is speculated that the temperature was high and pigment substances were unstable in the process of decocting.The average transfer rates of other components were 50%～70%,and the average paste yield of standard decoction was 28.91%.20 peaks were transmitted from GF standard decoction to dispensing granules,the average content of components in the dispensing granules was significantly different from that in the standard decoction.The average content of geniposide was 31.78%of that in the standard decoction,the value of hydroxygardenoside and crocin Ⅰ were 13.20%and 14.03%,but the value of deacetyl asperulosidic methyl ester was as high as 85.85%,It was suggested that the related components are further transformed in the preparation process of formula particles due to heating and drying.Cluster analysis showed that GF dispensing granules and standard decoction could be clustered into four groups,two groups of granules and two groups of standard decoction.The results of principal component analysis and factor molecular comprehensive score showed that the quality of GF dispensing granules was inferior to that of standard decoction.(3)According to HPLC fingerprint,there were 32 peaks on RRR decoction pieces and 28 peaks on RRR standard decoction.The transfer rate was 87.50%.26 components in the common peak were confirmed by comparision of the reference substance,which were gallic acid-4-O-β-D-glucoside,gallic acid,atechin,epicatechin,epicatechin gallic acid ester,2-O-cinnamoyl-β-D-glucoside,aloe emodin-8-O-glucopyranoside,resveratrol 4’-O-β-D-(6"-O-gallinoyl)-glucoside,rhein-8-O-glucopyranoside,l-O-gallinoyl-2-O-cinnamoyl-β-D-glucoside,sennoside A,1,6-di-O-galloyl-2-O-cinnamoyl-β-D-glucoside,emodin-1-O-glucopyranoside,1,2-di-O-galloyl-6-O-cinnamoyl-β-D-glucoside,6-methoxyacid-8-O-β-D-glucoside,chrysophanol-1-O-glucopyranoside,chrysophanol-8-O-glucopyranoside,emodin-8-O-glucopyranoside,aloe emodin-3-CH2-O-glucopyranoside,physcion-8-O-glucopyranoside,aloe emodin,rhein,emodin,chrysophanol,physcion.By comparing the mass spectrum data with the literature and the data of the compound library,six new compounds were inferred.The ratio of the common peak area of standard decoction and decoction pieces showed a good positive linear relationship with the component transfer rate.The average transfer rate of gallic acid was exceeded 100%,which was 233.00%.The average transfer rate of free anthraquinone was low.The average transfer rates of physcion,chrysophanol,emodin and aloe emodin were 1.41%,1.90%,3.66%and 13.70%,respectively.However,the average transfer rates of rhein,5 free anthraquinone,8 combined anthraquinone were 37.31%,11.03%and 44.04%,respectively.It was speculated that the transformation of components occurs,and anthraquinones makes the transition into tannins during the preparation of standard decoction.and the average paste yield of standard decoction was 23.26%.23 peaks were transmitted from RRR standard decoction to dispensing granules,the average content of components in the dispensing granules was signifieantly different from that in the standard decoction.The average content of rhein was 33.41%of that in the standard decoction,but the ratio of the other four free anthraquinone was more than 100%,the maximum of physcion was 366.86%,and the ratio of the five free anthraquinone was 64.29%,the reason was that the content of rhein in the standard decoction was relatively high.Besides aloe emodin-3-hydroxymethyl-O-glucopyranoside,the ratio of other 7 components were more than 100%,and the ratio of the 8 components was 147.42%;the average ratio of gallic acid and catechin,2 tannins,were 34.32%and 74.07%,respectively.It was suggested that the related components are further transformed in the preparation process of formula particles due to heating and drying.The results of cluster analysis showed that RRR dispensing granules and standard decoction could be grouped into two groups,but there were some intersections between dispensing granules and standard decoction.The results of principal component analysis and factor molecular comprehensive score showed that the quality of rhubarb dispensing granules was inferior to that of standard decoction.