| Yanyan Tablets, which is composed of twelve herbs:Radix Scrophularia, Cortex Moutan, Radix Rehmanniae, Semen Oroxyli, Radix Isatidis, et al, was listed in the Drug Specifications Promulgated by the Ministry of Public Health, P R China. Vol.2. It has been used to treat sore, anti-inflammatory actives, and effection of the peogaster system disease in the thesis. In this study, RP-HPLC method was established to determine the contents of the major active constituents in Yanyan tablets, and the HPLC-FPS was achieved for the 10 batches of tablets samples. The UPLC-MS/MS method was fully applied for simultaneous determination of harpagoside and cinnamic acid in rat plasma and investigate pharmacokinetic parameters of after oral administration of Yanyan tablets.A high performance liquid chromatography (HPLC) method has been developed for the determination of harpagoside, cinnamic acid, paeonol, rutoside and baicalin in Yanyan tablets. A Kromacil C18 column was adopted, the mobile phase was methanol-water(contained 0.05% phosphoric acid) for gradient elution with flow rate of 1.0 mL-min-1. The detection was set at 278 nm, the column temperature was maintained at 35℃. Good linearities were obtained from 0.200 to 4.00,0.208 to 4.16,0.610 to 12.2,0.960 to 19.2,1.00 to 20.0μg-mL-1 for harpagoside, cinnamic acid, paeonol, rutoside and baicalin, respectively. The mean recoveries of components were 100%, 102%,101%,98.3%,101%, respectively. The method is simple and accurate with good reproducibility. It has been successfully applied to the quantitave determination of the main components of Yanyan tablets.An HPLC-FPS of Yanyan tablets was analyzed. The method was performed on a Zirchrom Kromasil C18 column with a mobile phase of methanol-water(contained 0.05% phosphoric acid) for gradient elution at a flow-rate of 1.0 mL-min-1. The detection was by UV absorption at 280 nm and column temperature was 30℃. With cinnamic acid as reference substance, there were 16 common peaks in the fingerprints.10 batches of Yanyan tablets were identified with hierarchical cluster analysis and divided into two classes. According to the results of classification, common model was established by the better quality group and similarities were determined from 0.431 to 0.969 by using Similarity Evaluation System for Chromatographic Fingerprint of TCM of the Chinese Pharmacopoeia Committee. The result indicated that the similarity was different significantly. The developed method offered guarantee for standardizing the sources of herbs and the stability of the preparation technology and it was beneficial to improving its quality and the clinical activity and safety could be ensured.A sensitive, specific method has been developed for simultaneous determination of harpagoside and cinnamic acid in rat plasma using liquid-chromatography tandem mass spectrometry (UPLC-MS/MS). With hesperidin as the internal standard (IS), plasma samples were prepared by protein precipitation with methanol. The analysis was carried out on an ACQUITY UPLCTM BEH C18 column using acetonitrile-5 mmoL ammonium acetate (35:65, v/v) as the mobile phase. The detection was performed by means of electrospray ionization mass spectrometry in negative ion mode with multiple reaction monitoring (MRM). Linear calibration curves of harpagoside and cinnamic acid were obtained over the concentration ranges of 8.0-960 ng/ml and 9.9-992 ng/ml respectively. The intra-and inter-day precisions(RSD) were within 9.3% and 8.9% for harpagoside and 8.4% and 8.4% for cinnamic acid. The accuracy(RE) was from-4.2 to 4.6% for harpagoside and from 2.6 to 6.5% for cinnamic acid. The recoveries were all above 80%. The validated method was successfully applied to the pharmacokinetic studies of harpagoside and cinnamic acid in rats after oral administration of Yanyan tablets.The Cmax of harpagoside and cinnamic acid were 128±31 ng-mL"1 and 323±77 ng-mL-1, respectively, the Tmsx. were 0.50 h and 0.39 h, the t1/2 were 3.31 h and 7.31 h, and the AUC0â†'∞were 442.7±89.7 ng-mL"1·h and 1741±418 ng-mL-1-h, respectively. harpagoside and cinnamic acid appeared to be absorbed fast and eliminated slowly in vivo. |
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