Study On Components Analysis Of Yinchenhao Decoction And Quality Evaluation Of Its Prescription Pieces

Objective:Comprehensively analyze the chemical constituents in Yinchenhao Decoction(YCHD),and establish the quality evaluation of Artemisiae Scoparia Herba(ASH),Gardeniae Fructus(GF),and Rhei Radix et Rhizoma(RRR),which lays a foundation for the follow-up development of new formulations of YCHD.Methods:(1)Ultra-fast liquid chromatography-quadrupole time-of-flight mass spectrometry(UFLC-Q-TOF-MS/MS)was used to determine the components of YCHD.The components of YCHD were identified by comparison of reference substances,literature comparison,accurate molecular weight search and fragment ion analysis;(2)Thirty-five batches of ASH,36 batches of GF and 35 batches of RRR pieces were determined by high performance liquid chromatography.Establish the control fingerprints,calibrate the common peaks and evaluate the similarity.Components of common peaks were identified by UFLC-Q-TOF-MS/MS and the content of main components confirmed by reference substance was determined.Results:(1)A total of 451 ingredients were found and 347 of them were identified from YCHD.There are 23 ingredients from ASH,including 12 organic acids(among them,1-caffeoylquinic acid,1,3-Dicaffeoylquinic acid,1,5-Dicaffeoyl-quinic acid were confirmed by reference substance),eight flavonoids(among them,Quercetin-3-O-rhamnoside was confirmed by reference substance);two chromones and one other component(p-hydroxyphenyl was confirmed by reference substance).There are 92 ingredients from GF,including 35 iridoids(among them,geniposide,shanzhiside,geniposidic acid,Deacetyl asperulosidic acid methyl ester,gardenoside,Scandoside methyl ester,Genipin Gentiobioside were confirmed by reference substance),10 crocins(among them,crocin I and crocin II were confirmed by reference substance),15 organic acid,15 monocyclic monoterpene(among them,Jasminoside B were confirmed by reference substance),three flavonoids;seven triterpenoids;three other components,four tannins.There are 216 ingredients from RRR.including 49 anthraquinones(among them,eight bound anthraquinones of aloe-emodin-8-O-glucoside,rhein-8-O-glucoside,emodin-1-O-glucoside,chrysophanol-1-O-glucoside,chrysophanol-8-O-glucoside,aloe emodin-3-hydroxymethyl-O-glucoside,emodin-8-O-glucoside,physcion-8-O-glucoside and five free anthraquinones of aloe-emodin,rhein,emodin,chrysophanol,physcion were confirmed by reference substance),71 anthrones(among them,sennoside B and sennoside A,confirmed by reference substances),54 tannins(among them,gallic acid,catechin,epigallocatechin gallate,epicatechin and epicatechin gallate were confirmed by reference substance),nine butyrophenone[among them,4-(4’-hydroxyphenyl)-2-butanone 4-O-β-D-(2"-O-galloyl-6"-O-p-hydroxy-cinnamyl)glucoside,confirmed by reference substances],14 stilbenes[among them,resveratrol 4’-O-glucoside and resveratrol 4’-O-β-D-(6"-O-galloyl)-glucoside,confirmed by reference substances],eight chromones,two flavonoids,one organic acid and eight other components.There are 11 common ingredients from ASH and GF,including eight organic acid components(among them,chlorogenic acid,neochlorogenic acid,cryptochlorogenic acid,caffeic acid,3,4-Dicaffeoylquinic acid,3,5-Dicaffeoylquinic acid,4,5-Dicaffeoylquinic acid,confirmed by reference substances),three flavonoids(rutin,hyperoside and isoquercitrin,confirmed by reference substances).There were two common ingredients from ASH and RRR,including one flavonoids and one other components.There are three common ingredients from ASH,GF and RRR,including two flavonoids(Quercetin and Kaempferol 3 rutinoside,confirmed by reference substances)and one other components.There were 104 unidentified components,including 45 from ASH,23 from GF,20 from RRR,five common ingredients from ASH and GF,two common ingredients from ASH and RRR,and nine common ingredients from ASH,GF and RRR.(2)There were 42 common peaks in the fingerprint of 35 batches ASH pieces,and 23 of the 42 peaks were identified by comparison of reference materials,literature comparison and mass spectrometry,including 12 organic acids(among them,1-caffeoylquinic acid,neochlorogenic acid,chlorogenic acid,caffeic acid,cryptochlorogenic acid,1,3-dicaffeoylquinic acid,3,4-dicaffeoylquinic acid,3,5-dicaffeoylquinic acid,4,5-dicaffeoylquinic acid,confirmed by reference substances),10 flavonoids(among them,rutin,hyperoside,isoquercitrin,kaempferol-3-O-rutinoside,quercetin-3-O-rhamnoside,confirmed by reference substances),one other component(p-hydroxyphenyl,confirmed by reference substance).Thirteen components(including 1-caffeoylquinic acid,neochlorogenic acid,chlorogenic acid,caffeic acid,cryptochlorogenic acid,1,3-Dicaffeoylquinic acid,3,4-Dicaffeoylquinic acid,3,5-Dicaffeoylquinic acid,rutin,hyperoside,isoquercetin and p-hydroxyphenylacetone)in 35 batches of ASH pieces was quite different.(3)There were 60 common peaks in the fingerprint of 36 batches GF pieces,and 45 of the 60 peaks were identified by comparison of reference materials,literature comparison and mass spectrometry,including 21 iridoids(among them,geniposide,shanzhiside,geniposidic acid,deacetyl asperulosidic acid methyl ester,gardenoside,scandoside methyl ester,genipin gentiobioside were confirmed by reference substance),four crocins(among them,crocin I and crocin II were confirmed by reference substance),six organic acid(among them,chlorogenic acid,cryptochlorogenic acid,caffeic acid,4,5-Dicaffeoylquinic acid,confirmed by reference substances),three flavonoids(rutin,isoquercitrin and quercetin,confirmed by reference substances);8 monocyclic monoterpene(among them,Jasminoside B were confirmed by reference substance),one other component.16 components(including geniposide,shanzhiside,geniposidic acid,deacetyl asperulosidic acid methyl ester,gardenoside,scandoside methyl ester,genipin gentiobioside,Jasminoside B,crocin I and crocin II,rutin,isoquercitrin,chlorogenic acid,cryptochlorogenic acid,caffeic acid,4,5-dicaffeoylquinic acid)in 36 batches of GF pieces was quite different.(4)There were 45 common peaks in the fingerprint of 35 batches RRR pieces,45 peaks were identified by comparison of reference materials,literature comparison and mass spectrometry,including including 17 anthraquinones(among them,eight bound anthraquinones of aloe-emodin-8-O-glu-coside,rhein-8-O-glucoside,emodin-1-O-glucoside,chrysophanol-1O-glucoside,chrysophanol-8-O-glucoside,aloe-emodin-3-hydroxymethyl-O-glucoside,emodin-8-O-glucoside,physcion-8-O-glucoside and five free anthraquinones of aloe-emodin,rhein,emodin,chrysophanol,physcion were confirmed by reference substance),two anthrones(sennoside B and sennoside A,confirmed by reference substances),17 tannins(among them,gallic acid,catechin,epigallocatechin gallate,epicatechin and epicatechin gallate were confirmed by reference substance),two stilbenes[resveratrol 4’-O-glucoside and resveratrol 4’-O-P-D-(6"-O-galloyl)-glucoside,confirmed by reference substances],four butyrophenone[among them,4-(4’-hydroxyphenyl)-2-butanone4-O-β-D-(2"-O-galloyl-6"-O-p-hydroxycinnamyl)glucoside,confirmed by reference substances],two chromones and one naphthalenes.The content of 13 anthraquinones and 10 non-anthraquinones in 35 batches of RRR pieces was quite different.Conclusion:There are more than 300 components in YCHD,including organic acids and flavonoids from ASH,iridoid terpenoids and organic acids from GF,anthraquinones,anthrones and tannins from RRR,and various components in ASH,FD and RRR pieces was quite different.