Coptis - The Evodia Drug To The Dissolution Of The Main Component Of Change And Drug Action Studies

The pharmacologic actions of coptis-evodia herb couple changed with the proportion of each herb,but the chemical foundation and involved mechanisms have not been revealed.The thesis investigated the chemical composing of the herb couple and their dissolution rates in vitro and the fate in vivo by applying liquid chromatography-mass spectrometry(LC-MS)and high performance liquid chromatography(HPLC).It would offer some references for elucidating the mechanisms of coptis-evodia herb couple based on chemical composing.1.Study on the chemical composing of coptis-evodia herb coupleThe Semi-bionic Extraction was optimized by homogenous design while the score of total area of 17 peaks together with the quantity of berberine was chosen as marker. The optimized conditions as follows:pH values for three extraction processes were 1.6, 6.0,7.0 respectively and total extraction time was 2 h.A comprehensive HPLC-DAD-MS method was developed to study the chemical composing of semi-bionic extracts of coptis-evodia herb couple.The extract was separated on a Hypersil BDS C₁₈column(4.6 mm×200 mm,5μm,Elite,China)using acetonitrile-ammonium formic buffer as mobile phase by gradient elution.Detection was performed on DAD and MS equipped with an electrospray ionization(ESI)source by full scan and product full scan on positive mode.The chromatogram of coptis-evodia showed 17 main peaks,eight of which were from evodia while the others were from coptis,and it suggested that no new component was created in the semi-bionic extract of the herb couple.By comparison of the retention time,the on-line UV spectra and MS spectra,four peaks were identified as jatrorrhizine,hydroxevodiamine,palmatine and berberine,and three peaks were deduced as epiberberine,columbamine and coptisine.Seven samples with different proportions of coptis and evodia(1:0,1:1,1:2,1: 6,6:1,2:1,0:1)were extracted and then analyzed by RP-HPLC,in order to study the rule of solubility changes with the content of the copfis-evodia herb.Seventeen components were investigated in our experiment.Dissolution rate of the components from evodia elevated with the increasing of coptis;nine components were from coptis, three of which increased while the others decreased with the increasing of evodia.The proportion had different influence on the rule of solubility of different components in the herb.2.Pharmacokinetic studies of main components from coptis after oral administration of coptis-evodia herb couple in ratsA rapid and sensitive liquid chromatography/tandem mass spectrometry (LC-MS/MS)method was developed to simultaneously determine berberine,palmatine and jatrorrhizine in rat plasma.After mixing with the internal standard(IS) tetrahydropalmatine,plasma samples were pretreated by protein precipitation. Chromatographic separation was carried out on a Hypersil BDS C₁₈column using a mixture of water(containing 0.1%formic acid)and acetonitrile(30:70,v/v)as mobile phase.Detection was performed by selected reaction monitoring(SRM)mode via electrospray ionization(ESI)source operating in the positive ionization mode.The method was linear over the concentration range of 1-250 ng/mL for all components. The fully validated LC-MS/MS method has been successfully applied to the pharmacokinetic studies of berberine,palmatine and jatrorrhizine in rat plasma after oral administration of coptis-evodia(6:1,g/g)herb couple.Three peaks were observed in both individual and mean plasma-concentration curves of berberine,palmatine and jatrorrhizine.The concentration-time data was analyzed by non-compartmental methods in order to obtain the pharmacokinetic parameters:C_maxfor berberine,palmatine and jatrarrhizine is 121.1±11.9 ng/mL,35.2±9.5 ng/mL and 219.9±12.8 ng/mL, respectively.AUC_0-∞for berberine,palmatine and jatrarrhizine is 28341.5±5241.2 ng·min/mL,6345.5±133.9 ng·min/mL and 43576.9±4767.8 ng·min/mL,respectively. t_1/2for berberine,palmatine and jatrarrhizine is 336.9±46.5 min,215.7±33.5 min and 325.3±8.0 min,respectively,t_maxis 90 min for each component.3.Presumption of the metabolites of main components from coptis after oral administration of coptis-evodia herb couple in ratsThe present study characterized the metabolic profile of main components from coptis after oral administration of coptis-evodia herb couple in rats by analyzing plasma samples before and after dosing with the aid of LC-MS/MS.The phaseâ… metabolic position and pathways were supposed as O-dealkylation.The phaseâ…¡metabolites were identified as glucuronide conjugate of jatrorrhizine and phaseâ… metabolites.But it was a kind of supposition for lack of further validation and absence of compounds.