Screening anti-inflammatory components from Chinese traditional medicines using a peritoneal macrophage/cell membrane chromatography-offline-GC/MS method

We report the development of an analytical method combining cell membrane chromatography (CMC) with gas chromatography/mass spectrometry (GC/MS). This was applied to the purification and identification of anti-inflammatory components from traditional Chinese medicines. The stationary phase of the CMC employed mouse peritoneal macrophage (PM) cell membranes. We investigated the performance of the PM/CMC-offline-GC/MS method using hydrocortisone (HC) and dexamethasone (DM) as standards. The method was then applied to the identification of anti-inflammatory components in extracts of Rhizoma Atractylodes macrocephala (RAM) and Rhizoma Atractylodes lancea Thunb DC (RALD). The major components from both species retained by CIVIC were identified as atractylenolide I (AO-I) by GC/MS. Competition experiments' results showed that AO-I and lipopolysaccharide (LPS) bound competitively to cell surface receptors while AO-I and HC had only partly overlapping binding sites on the PM membrane. In vitro experiments revealed that AO-I was able to inhibit LPS-induction of TNF-alpha, IL-1 beta and NO production in a close-dependent manner. IC50 values were 5.3 mu g/mL, 5.1 mu g/mL and 7.5 mu g/mL, respectively. The PM/CMC-offline-GC/MS method is an effective screening system for the rapid detection, enrichment, and identification of target components from complex samples. (C) 2009 Elsevier B.V. All rights reserved.