RESUMEN
Objective: The aim of the present study was to determine the quality marker (Q-Markers) of Sparganii Rhizoma against thrombus through an integration of investigations on its antithrombotic effect, content determination and spectrum-effect correlation analysis. Methods: Based on the concept of Q-Marker, Sparganii Rhizoma was investigated for the identification of chemical component. The pharmacological effects on arachidonic acid-induced thrombosis in zebrafish were also investigated. The material basis in ethanol extract was determined by HPLC-UV. Furthermore, the potential Q-Markers were analyzed and predicted according to the effect-chemical correlation analysis. Finally, the anti-thrombotic Q-Markers were verified through the anti-thrombotic test of monomer components. Results: The model of thrombosis zebrafish was established with larvae exposed to 100 µmol/L arachidonic acid for 1 h. Nine ingredients in Sparganii Rhizoma were identified as 5-hydroxymethylfurfural, vanillic acid, ferulic acid, p-hydroxybenzaldehyde, p-hydroxybenzoic acid, vanillin, protocatechuic acid, p-coumaric acid and isoferulic acid. According to the determination effect of zebrafish thrombosis model and HPLC content analysis results, all the other contents present positive correlation except 5-hydroxymethylfurfural, and the P values of three representative potential Q-Markers (ferulic acid, protocatechuic acid and p-coumaric acid) were 0.002, 0.001 and 0.026, respectively. Conclusion: Sparganii Rhizoma showed a dose-dependent effect on the recovery of reducing cardiac red blood cell on zebrafish model. Three phenolic acids (ferulic acid, protocatechuic acid and p-coumaric acid) were proved to possess the anti-thrombotic effects which could be regarded as the potential Q-Markers for quality assessment of Sparganii Rhizoma.
RESUMEN
Objective: To optimize the extraction technology of two major iridoid glucosides (specnuezhenide and nuezhenoside G13) from four kinds of Osmanthus fragrans (OF) seeds, and to evaluate the anti-thrombotic activity of OF seeds. Methods: The orthogonal-test experiment was employed to optimize the parameters including ethanol concentration, liquid-material ratio, and extraction time for three extraction methods (ultrasonic extraction, reflux extraction, and microwave extraction). The extraction yield, content, and total peak area of iridoid glucosides were selected for weighted analysis to determine the best extraction method and technology. Additionally, an anti-thrombotic zebra fish model was established for biological evaluation of OF seeds. Results: Microwave extraction was the best method for iridoid glucosides extraction with the optimal conditions of ethanol concentration 55%, material-liquid ratio 1∶10, and microwave time 15 min. HPLC analysis showed that there was no significant difference in chemical composition among the four kinds of OF seeds. In zebra fish biological screening model, OF seeds displayed a weak inhibitory effect on the growth of thrombus and exhibited a pericardial edema effect in high dose-treated group. Conclusion: In this paper, extraction technology of two iridoid glucosides from four different kinds of OF seeds and preliminary anti-thrombotic activity evaluation of OF seeds were investigated. These results can provide the reference for further development and utilization of the agricultural waste of OF seeds.