ABSTRACT
Objective:To establish a high performance liquid chromatography (HPLC) method for determination of four active constituents, namely MSTG-A,MSTG-B,gaultherin and chlorogenic acid in the anti-inflammatory and analgesic active fraction (ARF) of the ethnic medicine Gaultheria leucocarpa var. yunnanensis, in order to provide a methodological basis for the in-depth study and quality control of G. leucocarpa var. yunnanensis,and lay a foundation for later preparation and clinical application. Method:The determination was performed on COSMOSIL 5C18-PAQ (4.6 mm×250 mm,5 μm) column with methanol-0.2% glacial acetic acid (gradient elution) as the mobile phase at a flow rate of 1 mL·min-1. The column temperature was 25℃. The detection wavelength was set at 294 nm. Result:The linear range of MSTG-B,MSTG-A,gaultherin and chlorogenic acid were 0.014 06-0.450 00,0.007 81-0.250 00,0.003 13-0.100 00,0.000 94-0.030 00 g·L-1 (r ≥ 0.999 7),respectively,with a good precision,repeatability and stability. And the average recoveries were 100.81%,98.99%,96.12% and 102.56%,respectively. RSDs were 1.4%,0.7%,0.7%,2.4%,respectively. The contents of MSTG-B,MSTG-A,gaultherin and chlorogenic acid in ARF fraction of G. leucocarpa var. yunnanensis were 23.608,41.973,8.282,2.673 mg·g-1,respectively. Conclusion:The established method was simple and accurate, with a high repeatability. It can be used for determination of four active constituents in ARF fraction of G. leucocarpa var. yunnanensis,so as to provide a reference for the in-depth research,quality control and comprehensive evaluation of G. leucocarpa var. yunnanensis and lay a solid foundation for preparation and clinical application.
ABSTRACT
Objective The active ingredient was used as index to optimize the extraction and enrichment process of anti-in-flammatory and analgesic active-fraction(ARF)of Dianbaizhu. Methods Methyl salicylate triglycoside-B was chosen as index com-ponent to extract and enrich methyl salicylate glycosides. Extraction and elution solvents were optimized. The HPLC fingerprint was ob-tained with Thermo Hypersil Gold C18(250 mm×4.6 mm,5μm)column and a gradient elution with the mobile phase consisting of ace-tonitrile(A)-0.2％acetic acid(B)at a flow rate of 1.0 ml/min. And the detection wavelength was set at 294 nm. Results The opti-mized extraction solvent of Dianbaizhu was the 30％ethanol and the optimized elution solvent of ARF enriched by AB-8 macroporous resins was the 35％ethanol. The methodological study on similarity and RSD in ARF HPLC fingerprint of three batches of samples cor-responded to related regulations. Conclusion The extraction and enrichment process of ARF is stable and repeatable.