Study On Effective Substance Of Callicarpa Nudiflora Hook.et Arn. And Design Of Preparation

Callicarpa nudiflora belong to Callicarpa genus,Verbenaceae family.It has the function of antibiosis,antiphlogosis and hemostasis.The herbal has exact Clinical efficacy and widely pharmacological effects.But the research is few on it’s chemical composition,the material basis of pharmacodynamics,pharmacokinetics,and pharmacy.The purpose of this study is to comprehensive reaserch on it’s active ingredients,pharmacodynamics,pharmacokinetics, pharmacy and so on,to provide the scientific basis for security application on clinic.GC-MS methods was used to analzye the constituents of essential oil of C.nudiflora.34 compounds were identified,which accounts for 87.28 percent of essential oil.The main chemical components were Humulene epoxide（17.28%）,α-Bisabolene epoxide（10.51%）, （+）-2-Carene（9.24%）,Terpinolene（8.70%） and Farnesol（6.65%）,etc.Six compounds were isolated from 80%ethanol extract of Callicarpa nudiflora by column chromatography,and identified as ursolic acid,β-sitosterol,Ayanin,apigenin,luteolin and quercetin.Orthogonal experiment design and Single-factor tests were used in optimizing the extraction and purification technology of total flavonoids in C.nudiflora（TFC） by measuring extraction Ratio.The main extraction process:the herbal were reflux-extracted by 10 multiples of 70%ethanol 2 times（2 hours per time）;The optimum technical conditions for purification of TFC by D101:diameter and high ratio of resin column:1:8,wet resin vs crude herbs:4:l,70%ethanol as eluting solvent,eluting rate at 2 mL/min,The yield of total flavonoids from leaves of C.nudiflora as this process was 3.41%,and the purity reached 74.57%.So the process is simple and convenient.The HPLC methods were developed to simultaneously determine quercetin,luteolin and Ayanin in TFC.Three components had a good linear relationship in the concentration of 19.19-95.96μg/mL,22.32-111.6μg/mL and 24.80- 124.0μg/mL and the recoveries were 98.5%,98.8%and 98.0%.The method about capillary permeability,the auricular tumefaction,the bleeding and blood-clotting time were used to investigate the efficiency of TFC on antiphlogosis and hemostasis .The results show that TFC have obviously inhibiting effects on the increase of mice’s abdominal capillary permeability caused by glacial acetic acid and the mice’s auricular tumefaction caused by dimethylbenzene;meanwhile it can obviously decrease the bleeding and blood-clotting time.The results of the antibiosis in vitro show that the MIC of EOC was 0.312%for Candida albicans and 0.625%for Staphylococccus aureus.The MBC of the oil was 0.625%for Candida albicans and 1.25%for Staphylococccus aureus.The HPLC-MS-MS methods were developed to determine luteolin,ayanin,and simultaneously determine these two compounds in rat plasma.The methods were sensitive,accurate and rapid.The method was applied to Phannacokinetic study of luteolin and ayanin after oral administration of TFC.The Parameters were calculated according to concentration-time curve by the principle of statistical moment.After oral administration of TFC,the relevant Parameters for luteolin and ayanin were Tmax 0.677h and 0.333 h;t_1/2 2.87h and 4.94 h;Cmax 380.3 andl.65ng/mL,respectively.The effect of every factors was investigated on encapsulation efficiency drug,loading capacity and release property of chitosan/alginate microspheres.The results showed that alginate concentration,ratio of alginate and TFC,chitosan concentration and CaCl₂ concentration had an effect on drug loading;encapsulation ration and release property of microspheres,reaction time had no effect on property of microspheres.The best prescription was that alginate concentration was 2.5%;ratio of alginate and TFC（A:D） was 3:1;chitosan was dissolved with 2%HAC,the concentration was 0.8%;CaCl₂ concentration was 2.0%; reaction time was 10min.The drug loading and encapsulation efficiency of microspheres respectively were 19.2%and 87.7%.EOC microemulsion was prepared.The effect of oil,surfactants and cosurfactant was investigated on the microemulsion.According to pseudo-ternary phase diagram,the formation was definited ethyl oleate and EOC（1:1） as oil phase,Tween80 as surfactant,in isopropyl alcohol as cosurfactant,distilled water as aqueous phase.EOC microemulsion gel was prepared with chitosan matrix.GC method was first established to determine terpinolene content in EOC microemulsion gel.Terpinolene had a good linear relationship in the concentration of 19.19 - 95.96μg/mL.