A simple, reliable, economical method was developed using HPLC with a diode-array detector for determination of total flavonoids in plasma after introvenous administration of ginkgo biloba extract. The method simultaneously detects quercetin, kaempferol, and isorhamnetin after acid hydrolysis and recalcula-tion. The hydrolysis and extraction conditions were optimized in an orthogonal test. The specificity was tested by comparing the retention time, UV spectra, and peak purity indices with standards. The detection limits were 20 ng/mL for quercetin, 20 ng/mL for kaempferol, and 50 ng/mL for isorhamnetin. The calibration curve ranges were 75-2400, 71-2280, and 70-2240 ng/mL. The pharmacokinetic characteristics of ginkgo biloba flavonoids after venous administration of 50 mg/kg ginkgo biloba extract to rats were analyzed using a two-compartment model. The initial plasma concentration was 171.22 μg/mL. The half-life of flavonoids in the first compartment (distribution) was 0.07 h and at the second compartment (elimination) was 4.51 h, while the AUC(0-∞) was 1711.06 μg·min/mL. The apparent volume of distribution was 0.11 L/kg. The total body clearance is 10.52 mL/(min·kg). The result shows the method is suitable for pharmacokinetic studies.
Ellagic acid (EA) has aroused great interest worldwide owing to its antioxidant, anti-inflammatory, and anticarcinogenetic properties. The EA content in pomegranate leaf was measured in this study using high performance liquid chromatography to investigate the effects of season, variety, and processing method on the EA level. The results show that the EA content in 11 varieties of pomegranate from the Zaozhuang region in China range from 1.30 mg · g–1 to 6.46 mg · g–1 of dry weight in five consecutive seasons from June to October. An analysis of variance (ANOVA) shows that the EA content is significantly dependent on the season (p<0.05). The EA content increases significantly during the growing season to the highest level in September and October. The effect of the leaf variety on the EA content is less significant than the season. The processing methods have different effects on the EA content. Soaking for 24 hours slightly increases the EA content (p<0.05). Heating at 80 ℃ or 100 ℃ for 1 h after soaking has little influence on the EA content, while slow-fired cooking at high temperature significantly elevates the EA content (p<0.05). To improve qual-ity and stability, several parameters such as leaf collection time, slow-fired cooking, and cooking time should be strictly controlled during the processing of pomegranate leaf tea and its extract.
