HPLC for Determination of Hyperoside in Rats Plasma and Its Application in Pharmacokinetics Study

OBJECTIVE To establish a HPLC for the determination of hyperoside (3-0-galactose-quercetin) in rats plasma and to study the pharmacokinetics of hyperoside in rats. METHODSBlood samples were collected via eyeball veniplex at different time intervals after injection of hyperoside through tail vein to rats at the doses of 12 mg·kg-1. The plasma protein was first precipitated directly with methanol and then the plasma concentrations of the hyperoside were analyzed by HPLC.A Shimadzu ODS column(4.6 mm×150 mm,5 μm)was used with a mobile phase consisting of methanol-water (40∶60).The detection wavelength was set at 360 nm. The mobile phase was run at a flow rate of 1.0 mL·min-1. The temperature of column was 25 ℃. Acetanilide was elected as the internal standard. RESULTSHyperoside and acetanilide were successfully separated. The linear range was 20-4 888 ng·mL-1, the regression equation of hyperoside was Y=132.539X-8.736,r=0.999 6. The precision of intra-assay and inter-assay was 0.1%-2.9% and 1.5%-7.3%. The average extraction recovery was 90.1% with RSD 0.9%. The results showed that the concentration-time curve of Hyperoside in rat plasma could be fitted to three-compartment open model and its pharmacokinetic parameters were as follows: t1/2(p)was (1.62±0.54)min,t1/2(a) was (18.67±2.48)min, t1/2(b) was (660.15±155.56)min, AUC was (200 229±4 397.63)ng·min·mL-1. CONCLUSIONThe HPLC method is simple and efficient with excellent specificity,sensitivity,precision,recovery and can be applied to pharmacokinetic investigation.