Studies On Pharmacokineics Of Oleuropein In Rats

Hepatitis B was the primary disease to threaten the health of human. Asthe Hepatitis B virus（HBV） get into liver cells, produce drug-fast cccDNAand long-term drugs to produce a single HBV, so far, there was not found agood drugs to fight HBV. The extraction, separation and synthesis of theactive component with anti-HBV in natural medicines became one of hotspots.Oleuropein was a phenol crack iridoid glycoside with anti-HBV, whichwas extracted from J．officinale. Oleuropein was rich in J．officinale,and waseasy to extracted. It was expected to be new lead compound to fight HBV. Butthere was a lack of the data of pharmacokinetics in vivo and the reported abortits metabolites in literature. Thus, further study on pharmacokineics ofoleuropein was demanded to carried out.The first part Study on the enrichment and separation of oleuropeinObjectiveTo enrich and separated oleuropein from J．officinalMethodsOleuropein was enriched and separated using polyamide resin, namely,the polyamide was carrier of separation, and the concentration of samplesolution was1g/mL, the sample volume was dry extract (g)/polyamide(g)=1:10, after the sample was static adsorption for2hours, it was washedwith distilled water3BV firstly, and then eluted using15%ethanol. Thecontent of oleuropein was detected by High Performance Liquid Chromatogra-phy (HPLC).ResultsThe sample was detected by High Performance Liquid Chromatography,The cotent of oleuropein was more then80% ConclusionThe sample was detected by High Performance Liquid Chromatography,the cotent of oleuropein was more then80%. It laid a foundation of furtherstudy on pharmacokineics of oleuropein in vivo.The second part Study on the choice of internal standard and chromatograp-hic condition in determination of oleurpein by HPLCObjectiveTo determine the internal standard and Chromatographic condition indetermination of Oleurpein by HPLC.MethodsTo scan the Oleurpein, baicalin, gallic acid, chlorogenic acid and otherreference material under the full wavelength by UV detector to determinedetection wavelength, using orthogonal design experiments, determined thechromatographic conditions, all the standard material retention time, peakwidth, calculated separation, according to the separation and retention time todetermine the optimum chromatographic conditions and internal standard.ResultsIn the orthogonal test, the column between the different levels F=39.50,between different levels of the mobile phase F=48.19, were greater than F0.05(2,2)=19, P <0.05, there was significant difference, but there was nosignificant difference between different standard material.ConclusionIn the orthogonal design, the oleurpein optimal chromatographicconditions Column: Agilent ZORBAX Eclipse XDB-C184.6×150mm5μm,mobile phase: water-methanol-formic acid60:40:1, internal standard wasbaicalin.The third part Study on the methodological investigation on phamrmacokin-etics of oleuropein in vivo of healthy ratsObjective To establish a HPLC method for the determination of oleuropein in rats’plasma.MethodsOleuropein was configurated into different concentration of standardsample, and then got into the blank plasma, after the protein was precipitatedby methanol–acetonitrile, the content of oleuropein in blank plasma wasdeterminated by HPLC.ResultsUnder the selected conditions, the peak shape of oleuropein was good.The linear range was5.25-31.5μg/mL, the lowest detection limitation was5.25μg/mL at s/N≥3. The precision of low, middle, and high concentrationplasma sample of oleuropein within and between days was good and RSD2.7%，9.7%，6.9%;3.1%，9.3%，8.7%, respectiveluy. The accuracy was-4.0%，7.5%，3.7%for low, middle, and high concentration, respectiveluy; Theextraction recovery rate of low, middle, and high concentration plasma sampleof oleuropein was95.9%，107.5%，100.4%; and low, middle, and highconcentration plasma sample of oleuropein was stable for3days.ConclusionThe method was sensitive, simple and selective, and was applicable to thedetermination of plasma concentrations and pharmacokinetic studies ofoleuropeinThe fouth part Study on pharmacokineics of oleuropein in ratsObjectiveTo study the pharmacokinetics of oleuropein in healthy rats.MethodsAfter the wistar rats were given orally three doses100,200,400mg/kg ofoleuropein, the blood samples were collected at different time, and then wascentrifμgated. After the protein was precipitated by methanol–acetonitrile,the content of oleuropein in rats’ plasma was determinated by HPLC. The dataof each group was analysised by3p97pharmacokinetic software. The pharmacokinetic parameters of each group were analysised using spss17.0softwwere.ResultsThe pharmacokinetics of oleuropein was best fitted with two-compartment model in rats after oral administration. There were significantdifferences in the values of t1/2, CL/F and AUC among the low, medium andhigh dose groups. The values of t1/2and CL/F were positively correlated withthe dose of medication. The values of AUC decreased but then increased whenthe dose of medication increased.ConclusionThe pharmacokinetics of oleuropein in rats were nonlinear process.