Studies On The Chemical Constituents Of Callicarpa Nudiflora And Quality Standard Of Callicarpa Nudiflora Capsules

Callicarpa nudiflora Hook belongs to the genus Callicarpa (Verbenacasefamily), which is mainly distributed in Hainan, Guangdong, and Guangxi Provinces.It is a famous region drug of Hainan Province. Growing in the ground to an altitudeof1200m hills, valleys, streams in the forest or bush. The roots, stems, leaves andflowers can be used as medicine. in addition, It have many benefits such asantiphlogistic, hemostatic, convergence, stasis detumescence, heat clearing anddetoxicating and carminative dampness. It is used as medicine for the treatments ofPulmonary Hemoptysis, gastrointestinal bleeding, epistaxis, Gum bleeding, traumaticbleeding, rheumatalgia and so on.Callicarpa nudiflora dried leaves were powdered and extracted with hot EtoH.The extract with petroleum ether, ethyl acetate and butyl alcohol extraction in turn.By multiple column chromatographies, reversed phase middle pressure liquidchromatography and HPLC preparation, twenty compounds were isolated from theextract. Their structures were determined by spectroscopic method. We finallydetermine nineteen compounds of them. They are composed of five flavonoids,(i)Luteolin (1), luteolin-4’-O-β-D-glucopyranoside (2), luteoloside (3), luteolin-3’-O-β-D-glucopyranoside (4), chrysoeriol-7-O-β-D-glucoside (5),(ii) two Catalpols: nudifloraCatalpol A (6),6’-O-caffeoyl-3,4-dihydrocatalpol (7),(iii) three Iridoids: nudifloside(8),7-O-E-p-coumaroyl-8-epi-loganic acid (9),7-O-E-p-coumaroyl-gardoside (10),(iv) one diterpenoid saponins:3,12-O-β-D-diglucopyranosyl-11,16-dihydroxyabieta-8,11,13-triene (11),(v) six triterpenoid saponins:2α,3α,19α,24-tetrahydroxyolean-12-en-28-oic-acid28-O-β-D-glucopyranoside (12),2α,3α,23,29-tetrahydroxyurs-12,19-dien-28-oic-acid28-O-β-D-glucopyranoside (13),2a,3a,19a,23-tetrahydroxyolean-12-en-28-oic-acid28-O-β-D-glucopyranoside (14),2α,3α,19α,23-tetrahydroxyolean-12-en-28-oic-acid28-O-β-D-xylopyranosyl-(1→2)-β-D-glucopyranoside (15),2α,3α,19α-trihydroxyolean-12-en-28-oic-acid28-O-β-D-xylopyranosyl-(1→2)-β-D-glucopyrano-side (16),2α,3α,19α-trihyhydroxy-urs-12-en-28-oic-acid28-O-β-D-xylopyranosyl-(1→2)-β-D-glucopyranoside (17),(vi) one phenylpropanoids: isoacteoside (18),(vii) one steroid: β-daucosterol (19). Among them, compounds6,7,9,10,12,13,15and16were new. Compounds5,11and14were isolated from Callicarpa for the firsttime.According to the characteristics of medication and the compounds isolated fromCallicarpa nudiflora which have gained results of pharmacological screening activeby the predecessors. Pharmacological results as follows： In the vitro assays,Compound6showed certain cytotoxic activity on HepG2and BGC-823two kinds oftumor cells with IC50values of3.37and2.49μM, respectively. In the invitro assays,Unfortunately, these Compounds rarely showed PTP1B inhibitory activity.Compounds3,6and18showed certain Alpha glucosidase inhibitory activity, withthe inhibition ratios31.7%,58.7%and35.8%. Compounds6,9,10,12,13,14and16showed pronounced hepatoprotective activities against D-galactosamine-inducedtoxicity in WB-F344rat hepatic epithelial stem-like cells.TLC was used in the identification of Callicarpa nudiflora Hook. Capsules withthe reference herb of Callicarpa nudiflora Hook. and HPLC was used for determinethe contents of luteoloside and acteoside. Analysis was performed on an DiamonsilC1（8250mm×4.6mm,5μm）and acetonitrile (A)-0.1%methanoic acid (B)(15:85)as the mobile phase, the flow rate was1.0mL/min, the detection wavelength ofluteoloside was set at350nm, the detection wavelength of acteoside was set at330nmand the column temperature was35℃. TLC spots were clear and well-separated.HPLC experiment showed good linear relationships at the ranges of2.26～90.24μg/mL (R2=0.9997) of luteoloside and8.90～356.2μg/mL (R2=0.9994) ofacteoside. The average recoveries (n=9) were99.57%(RSD=1.98%) and97.46%(RSD=1.32%), respectively. The method is simple and reliable with good precisionand it can be used for the quality control of Callicarpa nudiflora Hook. Capsules.We aim to quality control of studying on quality standard of Callicarpa nudifloracapsules for quality control, To ensure that the enterprises can produce stable quality,homogeneous and qualified products, so to give a guarantee of the safety for people.