Abstract
Pterocephalus hookeri, as a kind of popular traditional Tibetan medicine, is reputed to treat inflammatory related diseases. In the present work, a cyclooxygenase-2 functionalized affinity solid-phase extraction HPLC system was developed and combined with preparative-HPLC for rapidly screening and separating cyclooxygenase-2 ligand from P. hookeri extracts. Firstly, ligands of cyclooxygenase-2 were screened from extracts by affinity solid-phase extraction HPLC system. Then directed by the screening results, the recognized potential active compounds were targeted separated. As a result, the major cyclooxygenase-2 inhibitor of P. hookeri was obtained with a purity of >95%, which was identified as sylvestroside I. To test the accuracy of this method, the anti-inflammatory activity of sylvestroside I was inspected in lipopolysaccharide-induced RAW 264.7 cells. The results show that sylvestroside I significantly suppressed the release of prostaglandin E2 with dose-dependent, which was in good agreement with the screening result of the affinity solid-phase method. This method of integration of screening and targeted separation proved to be very efficient for the recognition and isolation of cyclooxygenase-2 inhibitors from natural products.
Highlights
Traditional Tibetan medicines (TTMs) are valuable for the treatment of plenty of clinical diseases in China [1]
In order to screen for active compounds with COX-2 inhibitory activity from P. hookeri and further explain its traditional use, a COX-2 functionalized the affinity solid-phase extraction HPLC system was developed to screen and characterize the cyclooxygenase-2 ligand from P. hookeri ethanol extracts pretreated via medium-pressure chromatographic tower
P. hookeri is rich in anti-inflammatory components, which were extracted by ethanol and enriched by microporous resin
Summary
Traditional Tibetan medicines (TTMs) are valuable for the treatment of plenty of clinical diseases in China [1]. Preparative HPLC (pre-HPLC), as a separation technique, is widely employed for purifying compounds with a high purity from the crude extraction of TTMs [28,29] This method is detected online with advantages of superior performance, excellent reproducibility and automation [30]. In order to screen for active compounds with COX-2 inhibitory activity from P. hookeri and further explain its traditional use, a COX-2 functionalized the affinity solid-phase extraction HPLC system was developed to screen and characterize the cyclooxygenase-2 ligand from P. hookeri ethanol extracts pretreated via medium-pressure chromatographic tower. The structure and the in vitro COX-2 inhibitory activity of the active compound are determined This advanced method, for the target separation of COX-2 inhibitors from complex extracts, could be applied for the isolation of ligands from natural productors in a highly efficient manner
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