Pharmacokinetic Study Of Alantolactone In Rat Plasma By HPLC Method And The Technology Study For Enzymatic Extraction Of Main Active Ingredient From Angelica

Elecampane and Angelica are commonly used in Chinese medicine herbs. As theprincipal constituent of Elecampane, alantolactone has been shown that have variouspharmacologic activities such as antiinflammatory and deworming. However, athorough literature search show that there is little information about thepharmacokinetics of alantolactone in vivo, which limits the clinical study andpraeparatum study of alantolactone. As clinical drug, Angelica's pharmacologicalactivity is already proved. But the particular or a certain type active ingredient ofAngelica are extracted based on boiled or with large amounts of organic solvent. Forthis reason, the utilization rate of medicine is lower and the active ingredient greatlylost. In addition, large amounts of organic solvents used in production process areharmful for environment. So, this subject is implemented from the following aspects:First of all, a HPLC method is developed for determination and pharmacokineticstudy of alantolactone in rat plasma following the administration of a singleintravenous (14mg/kg) or oral (30mg/kg) dose. After intravenous injection ofalantolactone, the concentration data in plasma is best fitted by two-compartmentopen models. The elimination half-life T_1/2β, body clearance (Cl_B) and area underthe concentration-time curve (AUC) are35.897±6.965min,0.00223(L/kg)/min and6.464±0.109g·min·L^-1, respectively. The elimination rate constant (K_e) is0.061±0.009min^-1. Following oral administration of alantolactone, the concentration data inplasma is best fitted by one-compartment open models. The elimination half-life (T1/2),_maximum plasma concentrations (C_max) and time of C_max(T_max) are44.999±7.017min,268.723±15.261μg·ml^-1and31.385±9.643min, respectively. The resultsshow that the obtained knowledge with the significant advantage can be used as asuitable reference in farther study of alantolactone.Secondly, enzymatic extraction technology of main active ingredient fromAngelica is studied. The effects of enzyme dosage, enzymolysis temperature,enzymolysis time, enzymolysis pH value on the extraction rate of ferulic acid andpolysaccharide are studied by single-factor test. Then, the orthogonal experiment andscreeing of compound enzyme are performed to obtain the optimum extractiontechnology. The research combines the traditional water extraction method andenzymatic extraction technology. Through this technology, extraction rate of ferulicacid has almost reached the level of traditional ethanol extraction method, and mainactive ingredient of Angelica is extracted, which are ferulic acid, ligustilide and polysaccharides. This technology could accomplish the purpose of comprehensiveutilization of Angelica. Polysaccharide extracted alone from Angelica by enzymaticextraction technology, after single test and orthogonal experiment, the extraction rateincreased by7.52%over traditional methods. Antitumor activity of Angelicapolysaccharide is evaluated by MTT method in hela cells. The results indicated thatAngelica polysaccharide extracted by enzymatic extraction technology have antitumoractivity against hela cells.The third, Angelica endophytes are isolated from different tissue of plant,including root, stem, leaf and soil. Then, the correlation between Angelica endophytesand main active ingredient of Angelica is studied. The optimal cultivation conditionof sterile angelica seedlings is determinated and these sterile angelica seedlings areinfected by endophytes isolated from angelica. Then the correlation between mainactive ingredient and endophytes is studied by HPLC after14days. The resultsindicate that endophytic fungi and mixed culture of strains have promoting effect onactive constituents of angelica (ferulic acid and ligustilide). Under the action ofendophytic fungi, ligustilide production increase by6.6times, and ferulic acid yieldincrease by4.1times. The production of the active ingredient is inhibited by bacteriaand actinomycetes.