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

• Published in: Journal of chromatography. B, Analytical technologies in the biomedical and life sciences Vol. 877; no. 27; pp. 3019 - 3024
• Main Authors: Wang, Changhe, He, Langchong, Wang, Nan, Liu, Fang
• Format: Journal Article
• Language: English
• Published: Amsterdam Elsevier B.V 01.10.2009; Elsevier
• Subjects: Analysis; Analytical, structural and metabolic biochemistry; Animals; Anti-inflammatory; Atractylenolide I; Atractylodes - chemistry; Biological and medical sciences; Cell Membrane - chemistry; Cell Membrane - drug effects; Cell Membrane - metabolism; Cell membrane chromatography; Cell Survival - drug effects; Chromatography - methods; Drugs, Chinese Herbal - analysis; Fundamental and applied biological sciences. Psychology; Gas Chromatography-Mass Spectrometry - methods; Gas chromatography/mass spectrometry; General pharmacology; Hydrocortisone - pharmacology; Interleukin-1beta - metabolism; Lactones - pharmacology; Macrophages, Peritoneal - chemistry; Macrophages, Peritoneal - drug effects; Macrophages, Peritoneal - metabolism; Medical sciences; Mice; Mice, Inbred BALB C; Nitric Oxide - metabolism; Peritoneal macrophage; Pharmacology. Drug treatments; Receptors, Cell Surface - metabolism; Reproducibility of Results; Rhizome - chemistry; Sesquiterpenes - pharmacology; Tumor Necrosis Factor-alpha - metabolism; Asia; China; Atractylenolide I; Cell membrane chromatography; Gas chromatography/mass spectrometry; Anti-inflammatory; Peritoneal macrophage; Terpenoid; Membrane chromatography; Pharmacognosy; Antiinflammatory agent; Lactone; Biomembrane; Screening; Folk medicine; Sesquiterpenes; Mass spectrometry; Macrophage; Peritoneum
• ISSN: 1570-0232; 1873-376X
• DOI: 10.1016/j.jchromb.2009.07.022

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 CMC 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-α, IL-1β and NO production in a dose-dependent manner. IC50 values were 5.3 μg/mL, 5.1 μg/mL and 7.5 μg/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.