| OBJECTIE:To establish a sensitive and specific method for determination of puerarin in rat plasma by HPLC and study its pharmacokinetics.METHODS:Puerarin was extracted from plasma with perchloric acid and was centrifugated by high speed. HPLC analysis was carried out through Hypersil ODS column( 4.6 mm×150 mm, 5μm ).The mobile phase consisted of methanol-0.0043mol·L-1 potassium dihydrogen phosphate ( 22:78 ) at a flow rate of 0.8 mL·min-1. The column temperature was 30℃and the wave length was 250 nm. Plasma puerarin concentration was determined by HPLC after intragastrically given of puerarin ( 100 mg·kg-1 ) or puerarin combined with berberine ( 50mg·kg-1, 100mg·kg-1, 200mg·kg-1 ) in rats. The data obtained were processed by DAS 2.1.1 software to calculate the varities of pharmacokinetic parameters after ig single durg or mixed drugs. RESULTS: 1. The standard curve was linear in the range of 0.10 mg·L-1～10.00 mg·L-1( A = 2.7019×104 C + 2495.8, r = 0.999 5 ). The lowest limit of detection was 0.05 mg·L-1. The relative standard derivation of inter-day and intra-day was smaller than 2.11% and 2.79%. The relative recoveries of Puerarine were 96.64%～112.55%.2. The main pharmacokinetic parameters of puerarin before and after coadministmtion of berberine were as follow: Cmax were 0.545±0.051 mg·L-1, 0.587±0.030 mg·L-1, 0.667±0.017 mg·L-1, 0.733±0.0298 mg·L-1; AUC0～t were 2.019±0.388 mg·L-1·h, 1.962±0.254 mg·L-1·h, 2.008±0.619 mg·L-1·h, 2.969±0.295 mg·L-1·h; CL were 32.937±12.695 L·h-1·kg-1, 32.656±3.599 L·h-1·kg-1, 34.556±9.167 L·h-1·kg-1, 22.748±4.654 L·h-1·kg-1. The results showed that, there was no significant difference in other pharmacokinetic parameters of puerarin between Group 1 and Group 2. While there were no significant differences for pharmacokinetic parameters of puerarin except increasing of Cmax after oral doses ( 100mg·kg-1 berberine, 200mg·kg-1 berberine ) ( P < 0.01 ). AUC0～t of puerarin increased in Group 4 compared with Group 1.CONCLUSIONS:The method was specific, sensitive and simple, and suitable for determination of Puerarin in rat plasma and pharmacokinetics researches. The results were accurate, a two-compartment pharmacokinetic model was adapted to Puerarin plasma concentration-time data analysis, the main pharmacokinetic parameters were similar to those reported domestic and abroad. So it could assist to approach internal compatibility in traditional Chinese medicine complex prescription.
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