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Han S.,Xian Jiaotong University | Han S.,National Prov Joint Engineering Research Center For Natural Vascular Medicine Screening And Analysis | Lv Y.,Xian Jiaotong University | Lv Y.,National Prov Joint Engineering Research Center For Natural Vascular Medicine Screening And Analysis | And 8 more authors.
Journal of Separation Science | Year: 2016

MaiLuoNing injection is a traditional Chinese medicine that used clinically since the 1950s in China. However, anaphylactic reactions, through the potentiation of mast cell degranulation, have been reported. In the present study, a rat basophilic leukemia-2H3 cell membrane chromatography coupled with high-performance liquid chromatography and electrospray ionization-ion trap-time of flight-mass spectrometry method was established for screening, analyzing, and identifying the potential anaphylactic components of MaiLuoNing injection. Harpagoside, a potential degranulator of rat basophilic leukemia-2H3 cells, was retained in rat basophilic leukemia-2H3 cell membrane chromatography. We aimed to evaluate the retained components to determine which of those were capable of inducing degranulation of basophilic leukemia cells. A β-hexosaminidase assay revealed that harpagoside can induce rat basophilic leukemia-2H3 cell degranulation in a dose-dependent manner. BLBA/c mice also exhibit passive cutaneous anaphylaxis in response to harpagoside. These results indicate that rat basophilic leukemia-2H3 cell membrane chromatography coupled with high-performance liquid chromatography and electrospray ionization ion trap time-of-flight mass spectrometry is effective in screening for the anaphylactic components of MaiLuoNing injection. © 2015 Wiley-VCH Verlag GmbH & Co. KGaA, Weinheim. Source


Han S.,Xian Jiaotong University | Han S.,National Prov Joint Engineering Research Center For Natural Vascular Medicine Screening And Analysis | Zhang T.,Xian Jiaotong University | Zhang T.,National Prov Joint Engineering Research Center For Natural Vascular Medicine Screening And Analysis | And 8 more authors.
Analytical Methods | Year: 2015

A highly sensitive high performance liquid chromatography-electrochemical detection (HPLC-ECD) method being used to detect released serotonin in real time from rat basophilic leukemia 2H3 (RBL-2H3) cells which can be stimulated by potential allergens was established to evaluate the sensitization of potential allergens. Chromatographic separation was carried out on a Chromolith R Speed ROD RP-18e column (50 mm × 4.6 mm I.D., 2 μm). The linearity of serotonin in samples was good, with correlation coefficients greater than 0.9986 within the corresponding concentration range. The relative standard deviation (RSD) percentages were in the range of 0.72%-2.96% for inter-day precision and in the range of 2.02%-3.48% for intra-day precision. The relative errors (REs) were within ±3.21%. The recovery of serotonin was in the range of 100.98%-101.56%, and the RSDs of recovery were less than 2.28%. The method allows for simple sample preparation, short analysis time, and high sensitivity, specificity, and reliability. This method was successfully used to detect serotonin released by RBL-2H3 cells stimulated by different concentrations of schisandrin A and harpagoside. The results demonstrate that schisandrin A and harpagoside can trigger RBL-2H3 cells to release serotonin in a dose-dependent manner. The study contributes to the further use of evaluation of potential allergens. © 2015 The Royal Society of Chemistry. Source

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