D Blood Ning Granules Quality Control Methods And Pharmacokinetics Study

Weixuening granules, included in Ministerial Standard of the People’s Republic of China was composed of eight Chinese herbs:Rhizoma et Radix Polygoni Cuspidat, Radix Paeoniae Alba, Herba Agrimoniae, Radix Rehmanniae, Caulis Spatholobi, Radix Rehmanniae Praeparata, Herba Ecliptae, Radix Pseudostellariae. It is widely used clinically to the treatment of thrombocytopenia and bleeding originated by heat in blood. In this paper, RP-HPLC was employed in quality control of index components in Weixuening granules, and an HPLC-UV method was developed for studying pharmacokinetics of polydatin contained in Weixuening granules in rats.Thirteen components in Weixuening granules were chosen for quality control. A RP-HPLC method was established to simultaneously determine the content of paeoniflorin, polydatin, verbascoside and resveratrol. The calibration curves were linear within the range of 22.3-223μg-mL-1 (r= 0.9999),61.1-611μg-mL"1 (r= 0.9997),6.38-63.8μg-mL"1 (r= 0.9996),4.88-48.8μg-mL"1 (r= 0.999 6) respectively, and the recoverise were 99.6% (RSD = 1.0%)、100.4%(RSD= 1.4%)、98.7%(RSD= 1.4%)、99.5%(RSD= 1.7%) respectively. Another RP-HPLC method was established to simultaneously determine the content of qercetin, lteolin, gnistein and formononetin. The calibration curves were linear within the range of 2.65-26.5μg-mL"1 (r= 0.9997),12.9-1.29μg-mL"1 (r= 0.9996),0.422-4.22μg·mL-1 (r= 0.9995),1.21-12.1μg-mL-1 (r= 0.999 6) respectively, and the recoverise were 99.3%(RSD= 1.9%)、100.1%(RSD= 1.8%)、98.7%(RSD= 1.7%)、99.5%(RSD= 1.1%) respectively. The third RP-HPLC method was established to simultaneously determine the content of catechin and epicatechin. The calibration curves were linear within the range of 3.33-33.3μg-mL-1 (r= 0.999 8),6.60-66.0μg·mL-1 (r= 0.999 7) respectively, and the recoverise were 101.4%(RSD= 1.6%)、98.0%(RSD= 1.1%) respectively. The forth RP-HPLC method were established to simultaneously determine the content of emodin and physcione. The calibration curves were linear within the range of 5.30-53.0μg-mL-1 (r= 0.999 8),0.988-9.88μg-mL-1 (r= 0.999 7) respectively, and the recoverise were 98.2% (RSD= 1.1%)、99.4%(RSD= 1.1%) respectively. The last RP-HPLC method was established to determine the content of catalpol. The calibration curve was linear within the range of 7.60-76.0μg-mL"1 (r= 0.999 7), and therecoverise were 98.1%(RSD= 1.4%) respectively.The method of determine the content of polydatin in rat plasma was developed by using HPLC-UV, and was used to study the pharmacokinetics of polydatin in rats after i.g. administration of Weixuening granules. The HPLC separation was achieved on a Diamonsil C18 (200 mm×4.6 mm,5μm) column with the mobile phase consisted of acetonitrile-water (20:80, v/v) at a flow rate of 1.0 mL·min-1. The column temperature was at 35℃. The UV detection wavelength was set at 303 nm. Linear calibration plot was obtained in the concentration range from 0.0986 to 9.86μg-mL-1 (r≥0.996 0). Intra-day RSD and inter-day RSD were both less than 7.7%. The extraction recovery of polydatin was exceeded 72.3%. The main pharmacokinetic parameters show this:Tmax was (0.403±0.063) h, Cmax was (1.715±0.097) g, ti/2βwas (2.484±1.624) h-1, AUC0→t was (1.511±0.550) mg·L-1·h, AUC0→∞was (1.955±0.765) mg·L-1·h. The pharmacokinetics of Polydatin was consistent with two-compartment model.