title: Analysis of allergens in tubeimu saponin extracts by using rat basophilic leukemia 2H3 cell-based affinity chromatography coupled to liquid chromatography and mass spectrometry.
abstract: An affinity two-dimensional chromatography method was developed for the recognition, separation, and identification of allergic components from tubeimu saponin extracts, a preparation often injected to treat various conditions as indicated by traditional Chinese medicine. Rat basophilic leukemia-2H3 cell membranes were used as the stationary phase of a membrane affinity chromatography column to capture components with affinity for mast cells that could be involved in a degranulation reaction. The retained components were enriched and analyzed by membrane affinity chromatography with liquid chromatography and mass spectrometry via a port switch valve. Suitability and reliability of the method was investigated using appropriate standards, and then, the method was applied to identify components retained from tubeimu saponin extracts. Tubeimoside A was identified in this way as a potential allergen, and degranulation assays confirmed that tubeimoside A induces RBL-2H3 cell degranulation in a dose-dependent manner. An increase in Ca(2+) influx indicated that degranulation induced by tubeimoside A is likely Ca(2+) dependent. Coupled with the degranulation assay, RBL-2H3 cell-based affinity chromatography coupled with liquid chromatography and mass spectrometry is an effective method for screening and identifying allergic components from tubeimu saponin extracts.
authors.affiliation: School of Medicine, Xi'an Jiaotong University, Xi'an, P. R. China.
authors.forename: Tao
authors.initials: T
authors.lastname: Zhang
authors.forename: Shengli
authors.initials: S
authors.lastname: Han
authors.forename: Qi
authors.initials: Q
authors.lastname: Liu
authors.forename: Ying
authors.initials: Y
authors.lastname: Guo
authors.forename: Langchong
authors.initials: L
authors.lastname: He
mesh_terms: D000485:Allergens / Q000737:chemistry*; D000818:Animals; D002118:Calcium / Q000737:chemistry; D002460:Cell Line; D045744:Cell Line, Tumor; D002462:Cell Membrane / Q000378:metabolism*; D002846:Chromatography, Affinity; D002851:Chromatography, High Pressure Liquid; D008407:Mast Cells / Q000166:cytology*; D008516:Medicine, Chinese Traditional; D010936:Plant Extracts / Q000737:chemistry; D051381:Rats; D015203:Reproducibility of Results; D012503:Saponins / Q000737:chemistry*; D053719:Tandem Mass Spectrometry; D014315:Triterpenes / Q000737:chemistry*; D001619:beta-N-Acetylhexosaminidases / Q000737:chemistry
chemical_list: D000485:Allergens; D010936:Plant Extracts; D012503:Saponins; D014315:Triterpenes; C051084:tubeimoside I; D001619:beta-N-Acetylhexosaminidases; D002118:Calcium
keywords: Cell membrane chromatography; Degranulation; High-performance liquid chromatography with mass spectrometry; Mast cells; Tubeimoside A