| 1. Determination of stilbene glucoside in rabbits and mice plasma by HPLC and study of its PharmacokineticesStilbene glucoside (2, 3, 5,4' -Tetrahydroxystilbene - 2-O--D-glucoside) , one of the water-soluble bioactive components in Radix polygon! multiflori, can reduce the total cholesterin in blood serum of rat. As a guide line it is used to control the quality of Radix polygoni multiflori in the Chinese Pharmacopoeia (2000). In order to clarify the pharmacokinetic characteristic of stilbene glucoside and to guide the use of clinical drug aright, we studied the pharmacokinetic parameters of stilbene glucoside using HPLC in rabbits and mice given drug intravenously and orally.Objective: To establish a method to determine the concentration of stilbene glucoside in plasma of mice and rabbits by HPLC. To study the phatmacokinetic parameters of stilbene glucoside that consists in Radix polygoni multiflori in the bodies of mice and rabbits.Method: The stilbene glucoside injection was injected into ear vein of rabbit. The dose was lOmg .kg-1. Blood samples (1.0ml)were collected from ear vein at 3, 4, 6, 8, 10, 12, 14, 16, 20, 25, 30, 40, 50, 60, 90and 120minpostdose. According to the prearranged time, mice were divided into different groups randomly. Every group had six mice. Some groups were injected with stilbene glucoside injection by taildifferent groups randomly. Every group had six mice. Some groups were injected with stilbene glucoside injection by tail vein. The dose was lOmg. kg-1. Blood samples were collected from vein or artery in the eyepit at 2, 3, 4, 5, 6, 8, 10, 12, 15, 20, 30, 45 and 60 min following the single dose of stilbene glucoside injection. The others were given the extract of Radix polygon! multiflori orally. The dose was 120mg. kg-1. At 2, 3, 4, 5, 8, 10, 15, 20, 30, 45, 60, 90, 120, 150 and ISOmin after oral administration , blood samples were taken from vein or artery in the eyepit .The blood samples were drawn into tubes with heparin sodium respectively. Plasma was obtained by centrifugation at approximately 3000 rpm for lOmin.Three-times volume of methanol was added and the content of tube was mixed to precipitate the plasma protein, then centrifiigedThe supematant(20 u l)was injected into the HPLC system. The HPLC consisted of a Diamonsil?Qg, 5 u m 250 X 4.6mm i.d. analytical column, a mobile phase of acetonitrile-methanol-l%formic acid (15:18:67), flow rate 1.0ml - min-1, UV detection at 320 run. The sensitivity was 0.5AUFS. The peak area of samples was used for predicting unknown concentrations form the regression equation. The date was handled with 3P97 pharrnacokinetics software. The best compartment model was chose by comparison between the observed concentrations and the concentrations predicted by the model. The Akaike Information Criterion (AIC) was used to indentify the best compartment model, too. The compartment model with the smallest AIC is the most adequate.Results: The mean plasma concentration-time curve of stilbene glucoside was determined after different administration of the stilbene glucoside injection to rabbits and mice. The standard curve range was 0.410-42.0 V- g. ml-1. The regression equations in rabbit and mouse are Y=1.0333X+0.2252, r=0.9999(n=8) and Y=1.0335XK).2189, r=0.9999(n=8), respectively. The lowest detection concentration was 0.0510 mg. ml-1. The recoveries in three different concentrations (high, middle and low) were 97.98%, 101.7% and 104.5%, respectively. The within-day precisions (RSD) were 8.7%, 2.9% and 5.5%, respectively. The mean plasma concentration-time curves of stilbene glucoside after intravenously administration were confirmed to open two-compartment model in rabbits and mice. The T1/2, T1/2 K21, K12, K10, V(c), V, AUC, CL of rabbits were 2.7min, 13.5min, 4.2h-1, 3.0h-1, 11.2h-1, 0.198L-kg-1,0.739L-kg-1,4.530mg-h-L-1,and 2.208L-h-1-kg-1, respectively. The T1/2, T1/2, K21, K12, K10, V(c) V, AUC, CL of mice were 1.9min, 8.3min, 6.6h-1, 3.8h-1 16.0h-1, 0.090L-kg-1, 0.289L.kg-1, 6.918mg. h. L-1 and 1.445L.h-1. kg-1, respect... |
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