| ObjectiveMesona chinensis Benth.belongs to Labiatae,namely "xianrencao","xiancao","dongcao","xincao" in Chinese.Quanlity evaluation of Mesona chinensis Benth.reported in literatuers were mainly concentrated in the aspect of content determination,but there are shortcomings,such as,one single index component,and the measured ingredient lack of specificity,quality control methods need to be further improved.Study on the chemical constituents of Mesona chinensis Benth.rarely reported,the lack of specific reference become a difficult problem to improve the quality control of herba Mesona.Polysaccharide content reached 43.84%,there has not been a determination of saccharide content in reports.Phenolic acids,flavonoids and sugars are the main component of herba Mesona,to a certain extent,the higher content of them,the better.To control the content of these components can ensure the quality of Mesona chinensis Benth..Therefore,in order to further understand the chemical composition of Mesona chinensis Benth.,ethyl acetate extraction,n-butanol extraction and water phase was studied.On the basis of the chemical component research,HPLC fingerprint qualitative method was builded to identify the authenticity of Mesona chinensis Benth.,and identified 8 characteristic peaks by comparing reference substances.In combination with principal component analysis results,the content of six major component was determined.The content of five sugar of Mesona chinensis Benth.by HPLC-ELSD was also determined in this paper,hope to provide basis for the establishment of its quality standard.Methods1.Ethyl acetate extraction,n-butanol extraction and water phase was separated succes-sively by using sillica column chromatography,ODS,Sephadex LH-20 and macroporous.NMR spectrum technique was used to identify the structure of compounds.2.HPLC fingerprint analytical method was established,chromatographic fingerprint of Traditional Chinese Medicine similarrity evluation software was used to evaluate the quality of 16 bathes of Mesona chinensis Benth.by SPSS.3.The content of lithospermic acid,salviaflaside,lithospermic acid B,rosmarinic acid,Salvianolic acid J and isoquercitin was determined by HPLC,and evaluated the quality of Mesona chinensis Benth.4.The content of fructose,a-D-glucopyranose,?-D-glucopyranose,sucrose,?-D-galactose-(?6)-a-L-glucose(1?2)-?-D-fructose was determined by HPLC-ELSD.Results1.42 compounds were identified,12 flavonoids:kaempferol-3-O-glucoside(LFC-1),keampferol-3-O-[6"-O-(trans-p-coumaroyl)]-p-D-glucopyranoside(LFC-2),kaempfer-ol-3-O-?-D-glucopyranoside-7-O-a-L-rhamnoside(LFC-3),kaempferol(LFC-4),keampf-erol-3,7-di-O-?-D-glucoside(LFC-5),kaempferol-7-O-?-L-rhamnoside-4'-O-?-D-glucopy-ranoside(LFC-6),isoquercitrin(LFC-7),quercetin-3-O-(6"-O-trans-p-coumaroyl)-?-D-glu-copyranoside(LFC-8),quercetin-3-O-?-D-(6-O-Z-p-coumaroyl)-glucopyranoside(LFC-9),quercetin-3-O-(6"-O-E-caffeoyl)-f3-D-glucopyranoside(LFC-10),quercetin(LFC-11),rutin(LFC-12);15 phenolic acids:rosmarinic acid(LFC-13),methyl rosmarinate(LFC-14),salviaflaside(LFC-15),salviaflaside methylester(LFC-16),salvianolic acid A(LFC-17),9',9"'-dimethyl lithospermate B(LFC-18),lithospermic acid B(LFC-19),methyl litho-spermate(LFC-20),lithospermic acid(LFC-21),caffeic acid(LFC-22),1-O-[(E)-caffeoy-1]-?-D-glucopyranose(LFC-23),protocatechuic acid(LFC-24),danshensu(LFC-25),salv-ianolic acid J(LFC-26),4-hydroxy-2,6-dimethoxyphenol-l-O-?-D-glucoside(LFC-27);9 sugar:fructose(LFC-28),2-methyl-a-D-fructofuranoside(LFC-29),2-methyl-?-D-fructof-uranoside(LFC-30),glucose(LFC-31),1-ethyl-?-D-glucopyranoside(LFC-32),2-methyl-?-D-fructopyranose(LFC-33),galactose(LFC-34),sucrose(LFC-35),?-D-galactose(1?6)-a-L-glucose(1?2)-?-D-fructose(LFC-36);3 triterpenoids:oleanolic acid(LFC-37),dau-costerol(LFC-38),2a,3?,19?,24-tetrahydroxyolean-12-ene-23,28-dioicacid-23-O-?-gluco-pyranoside(LFC-39);3 nitrogen compounds:3-indolecarboxylic acid(LFC-40),uridine(LFC-41),adenosine(LFC-42).2.11 common peaks in the HPLC fingerprint were determined,eight compounds were identified by comparing with reference substances:caffeic acid(1#),lithospermic acid(2#),isoquercitin(4#),salviaflaside(5#),kaempferol-3-O-glucoside(6#),lithospermic acid B(7#),rosmarinic acid(8#),salvianolic acid J(9#).The similarity result showed that 16 batches of herbs between 0.766?0.982,primary component analysis showed that the first principal component of lithospermic acid(2#),isoquercitin(4#),salviaflaside(5#),lithos-permic acid B(7#),rosmarinic acid(8#)and salvianolic acid J(9#)plays an important role in herbal materials classification.3.The content of six components in range:0.43?5.18 mg/g(lithospermic acid),0.29?1.79 mg/g(salviaflaside),3.47?22.34 mg/g(lithospermic acid B),1.60?7.35 mg/g(rosmarinic acid),0.36?2.40 mg/g(salvianolic acid J),0.16?2.56 mg/g(isoquercitrin),the average content was 2.05 mg/g,0.73 mg/g,11.00 mg/g,4.32 mg/g,1.10 mg/g,0.98 mg/g.The results showed that each batch of ingredients are quite different,w-ith an average content of lithospermic acid B was 11.00 mg/g,which was significa-ntly higher than the content of the other five compounds.According to the results,it was suggested that content limit of each compound was calculated by the average value of 80%,content of lithospennic acid is not less than 1.6 mg/g,content of sa-lviaflaside is not less than 0.6 mg/g,content of lithospermic acid B is not less than 8.8mg/g,content of rosmarinic acid is not less than 3.5 mg/g,content of salvianolic acid J is not less than 0.9 mg/g,isoquercitrin content of not less than 0.8 mg/g.4.The content of fructose,a-D-glucopyranose,?-D-glucopyranose,sucrose and ?-D-galactose-(1—>6)-a-L-glucose(1?2)-?-D-fructose was 3.60-16.86 mg/g,6.40-18.49 mg/g,6.45?19.81 mg/g,0.92?8.47 mg/g and 1.85?9.35 mg/g respectively,the average content was 10.06 mg/g,10.67 mg/g,12.47 mg/g,2.89 mg/g,4.26 mg/g respectively of Mesona chinensis Benth.by HPLC-ELSD.The average content of ?-D-glucose was the highest,and sucrose,,?-D-galactose-(1?6)-a-L-glucose(1?2)-?-D-fructose is slightly lower,the result showed that there are differences of each batch of component content,it also showed that the ?-D-glucose was the dominant configuration of the carbohydrate component.According to the results,it was suggested that content limit of each compound was calculated by the average value of 80%,content of fructose is not less than 8.0 mg/g,content of a-D-glucopyranose is not less than 8.5 mg/g,content of ?-D-glucopyranose is not less than 10.0 mg/g,content of sucrose is not less than 2.3 mg/g,content of ?-D-galactose-(1?)-?-L-glucose(1?2)-?-D-fructose is not less than 3.4 mg/g.ConclusionWe researched the chemical conponents of Mesona chinensis Benth.in this paper,42 compounds were identified,12 flavonoids,15 phenolic acids,9 sugars,3 terpenoids,3 nitrogen compounds.34 compounds were isolated from this plant for the first time,of which 33 compounds were isolated from the Mesona Bl..These chemical components are larger polar compounds.Flavonoids structure is based on quercetin and kaempferol,then deried from a series of compounds.Phenolic acid compounds is mainly composed of(C6-C3)of benzene propanoic acid derivative compositon,they have phenolic hydroxyl groups or styrenic structure.Sugar compounds with monosaccharides,methyl or ethyl monosaccharides,disaccharides and trisaccharides.In addition,we identified 3 nitrogen compounds(uridine,adenosine,3-indolecarboxylic acid).Monosaccharide,methyl or ethyl monosaccharide,disaccharide,trisaccharide and nitrogen compounds was found in this herba for the first time,which enriched chemical composition types and provided a reference for the later study.On the basis of chemical components,we established HPLC fingerprint,HPLC and HPLC-ELSD content determination method,comprehensive evaluation of the intrinsic quality of Mesona chinensis Benth. |
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