The Determation Of BuShenYiZhiFang's Intransitional Body Composition-osthole Content And Pharmacokinetics

ObjectiveRanging from the pharmacokinetics of chinese herbs, the subject starts to establish a method of measurating the quantity of Osthol in rabbit' serum by HPLC-UV method to study the drug Osthol-the main component of the drug in BuShenYiZhiFang, and calculate the pharmacokinetic parameters after gavage, then compare BuShenYiZhiFang with Cnidium decoction, to measure the quantity of osthol in cerebrospinal fluid after gavage in order to determine whether the active ingredient in BuShenYiZhiFang could go through the blood-brain barrier (BBB), whether the drug content of BuShenYiZhiFang in cerebrospinal fluid is higher than single decoction in it.MethodsNew Zealand rabbits were studied, using its blank serum, respectively, from a methodological specificity study, the lowest detection concentration, the preparation of the standard curve, precision and method recovery test, extraction recovery and stability of several aspects,a comprehensive testevaluation is established, by establishing a HPLC-UV method of determinating Osthol in rabbit' serum,drawing a drug-time curve and calculate its pharmacokinetic parameters by a software of Pksolution2.0 pharmacokinetic.AfterirrigatingBuShenYiZhiFangandcnidiumdecoction, determinate the drug content in cerebrospinal fluid, at the same time compare among the groups. Data results to indicate by"(?)±s ", using SPSS13.0 statistical software for statistical analysis.Results1 The establishment of HPLC-UV method in determinating Osthol in rabbit' serumThis study, using liquid-liquid extraction method for biological sample preparation, established a method of reversed-phase high performance liquid chromatography-UV detection, using a right-hydroxy acid as internal standard to determinate Osthol concentration in rabbit's serum. Its detection range, precision, recovery and stability, in line with biological sample analysis and pharmacokinetic study requirements. The Osthol concentration in serum is in the linear range of 0.052-3.333μg/mL, with the quantification limit of 0.013μg/mL.2 Calculate rabbit serum concentration after gavage, and the pharmacokinetic parameter.Pharmacokinetic study showed that rabbits given gavage of 0.18g/kg BuShenYiZhiFang drug, the pharmacokinetic behavior of osthol in body can be described in two-compartment model. Osthol drug peak concentration Cmax= 2.1 (ng/ml); Tmax= 30 (min); absorption phase half-life of t1/2ka= 21.103 (min), distribution phase half-life t1/2(α)= 19.985(min), elimination phase half-life t1/2(β)= 1095.601 (min), shows the absorption is fast, distribution is fast, and eliminating is relatively slow. Area under the curve AUC0→∞= 439.2 min Xμg /mL)3 The establishment of HPLC-UV method in determination of Osthol concentration in rabbit's cerebrospinal fluid.This experiment has established an accurate, reliable, stable and reproducible HPLC-UV method, in determination of Osthol concentration in rabbit's cerebrospinal fluid. It's essential for drug kinetics. In this study, through the chromatographic conditions, internal standard substance and extraction agents established to screen high-performance liquidchromatography and its detection range, precision, recovery and stability, in line with biological sample analysis and pharmacokinetic study requirements. TheOsthol concentration in cerebrospinal fluid is in linear range of 0.026-1.667μg/mL, with the quantification limit of 0.007μg/mL.4 Determination of the concentration of drug in cerebrospinal fluid after rabbits fed a compound, and a single drug.BuShenYiZhiFang Group and Cnidium water decoction group, differences between the two groups was statistically significant (P<0.01), fed with BuShenYiZhiFang Group osthol content in cerebrospinal fluid compared with fed Cnidium decoction CSF were significantly higher in osthol (P<0.01).ConelusionTo sum up, after fed BuShenYiZhiFang, its main active ingredient-osthol absorbed in the body faster, distribution is faster, and its through blood-brain barrier into the cerebrospinal fluid. The Osthol content of the compound group in the cerebrospinal fluid is more than single-drug groups, consider the reasons may be related to compound borneol open the blood-brain barrier for drug absorption.