Studies On The Chemical Constituents Of Callicarpa Nudiflora And Quality Standard Of Glechomae Herba

Glechomae Herba is one of the traditional Chinese medicine in China.We studied the chemical constituent of Glechomae Herba,set up the quality standard.It is of great significance to clarify the chemical compositions and establish quality standard of Glechomae Herba.And our study provided important scientific basis for making full use of toad skin of Glechomae Herba.In this dissertation,the following results were achieved:1.Study on the Chemical Constituents of Glechomae HerbaThe dried seeds of Glechomae Herba were powdered and extracted with 75%Et OH.The extract with dichloromethane,ethyl acetate and n-butanol extraction in turn.The extract liquor was systematically isolated and purified by multiple column chromatographies,reversed phase middle pressure liquid chromatography,TLC,and HPLC preparation.A total of eighteen compounds were isolated from the ethylacetate extraction,and their structures were elucidated by spectroscopic method and physical and chemical properties.Finally we determine fifteen compounds of them.They are composed of Rosmarinic acid（1）,methyl rosmarinate（2）,Salvianolic acid A（3）,caffeoylglycolic acid（4）,trichocarpin（5）,isorinic acid（6）,methyl-3,4-dicaffeoylquina te（7）,shimobashiraside C（8）,（E）-3-（（2S,3R）-3-（3,4-dihydroxyphenyl）-2-（（E）-3-（3,4-dihydroxyphenyl）acryloyloxy）-2,3-dihydrobenzo[b][1,4]dioxin-6-yl）acrylic acid（9）,Latifolicinin D（10）,eucommin A（11）,protocatechuic acid（12）,Apigenin-7-O-β-D-glucopyranoside（13）,Chrysoeriol-7-β-O-D-glucopyranoside（14）,[3-hydroxy-2-4-（4-hydroxyphenoxy）phenyl]-1-（methoxycarbonyl）ethyl（E）-3-（3,4-dihydroxyphenyl）prop-2-enoate（15）.A total of twelve compounds were isolated from the n-butanol extraction,and their structures were elucidated by spectroscopic method and physical and chemical properties.Finally we determine ten compounds of them.They are composed of.17-O-β-D-glucopyranosy-3-O-β-D-glucopyranosy-ent-pimara-8（14）,15-diene（16）,17-O-β-D-glucopyranosy（1→3）-β-D-glucopyranosy-3α-droxy-ent-pimara-8（17）,15-diene（17）、lucihirtin A（18）,ilexsaponin B₁（19）,12-methoxy-7-oxo-8,11,13-triene-3-O-β-D-glucopyranosyl-（1→2）-β-D-glucopyranoside（20）,12-met hoxy-7-oxo-8,11,13,15-tetraene-3-O-β-D-glucopyranosyl-（1→2）-β-D-glucopyranosid e（21）,2α,3α,23-trihydroxy-olean-12-en-28-O-β-D-glucopyranosyl-（1→6）-β-D-glucop yranosyl-（1→2）-β-D-glucopyranoside（22）,2α,3α,23-trihydroxy-urs-12-en-28-O-β-D-glucopyranosyl-（1→6）-β-D-glucopyranosyl-（1→2）-β-D-glucopyranoside（23）,3-O-β-D-glucopyranosyl-（1→3）-β-D-glucuronopyranosyl-21β-hydroxy-urs-12-en-28-al（24）,3-O-β-D-acetilglucopyranosyl-（1→3）-β-D-glucuronopyranosyl-21-O-β-D-acetilgluco pyranosyl-urs-12-en-28-al（25）.Among them,compounds 16、17 and 20～25were new.Compounds3～8、9、10、11、13、15、18 and 19 were isolated from Glechomae Herba for the first time.2.Study on the Quality Standard of Glechomae HerbaWe established an HPLC method to simultaneously determin of four phen olic acids chemical compositions in Glechomae Herba,namely,caffeic acid、caffeoylglycolic acid、rosmarinic acid and Salvianolic acid A,SAA.The CAPCE LL PAK C₁₈（250 mm×4.6 mm,5μm）was used with the mobile phase of ac etonitrile（A）-0.1%Formic acid（B）gradient elution,at the flow rate of 1.0 m L·min^-1,the detection wavelength was 330 nm,and the column temperature was at 30℃.Results:The linear range of caffeic acid、caffeoylglycolic acid、ros marinic acid and Salvianolic acid A,SAA were 0.034～0.85μg（r=0.999 8）、0.014～0.35μg（r=0.999 8）、0.524 5～13.112 5μg（r=0.999 8）and 0.008 4～0.21μg（r=0.999 9）,respectively.The average recoveries（n=3）were 98.2%～99.56%（RS D<0.72）﹑97.63%～99.48%（RSD<1.28）、100.2%～100.5%（RSD<0.91）and 98.08%～99.21%（RSD<1.02）,respectively.The contents of caffeic acid、caffeoylglycolic acid、rosmarinic acid and Salvianolic acid A,SAA in eight batches of samples were 0.238～1.121、0.036～0.180、1.461～11.011 and 0.042～0.124 mg·g^-1.Conclusion:The method is of high separationefficiency,good repeatability,the methodology validation,can be used to control the quality of the Glecho mae Herba.