Objective: To identify the constituents in Shuanghuanglian injection (SHLI) that correlate with anaphylactoid reaction.
Methods: Chemical fingerprints of 10 batches SHLI samples were determined by High Performance Liquid Chromatography (HPLC), and further investigated by similarity analysis. Combined with optical microscopy, both anaphylactoid experiments and confirmatory assay were displayed in Rat basophil leukemia cells (RBL-2H3) to obtain the histamine release inducing by SHLI. The content of histamine was tested by Enzyme-Linked Immuno Sorbent Assay method. Partial least squares regression (PLSR) method and HPLC-DAD-ESI-MSn technology were conducted to analyze constituents in SHLI involving anaphylactoid reaction.
Results: The results of spectrum and effect relationships showed that the eight constituents were positively correlated with anaphylactoid reaction. Among which, nearly 90% of them were identified as baicalin and rutin with PLSR and HPLC-DAD-ESI-MSn. This result was in accordance with confirmatory assay on RBL-2H3 cells.
Conclusion: Baicalin and rutin from SHLI were the main constituents involving anaphylactoid reaction.
Keywords: Anaphylaxis; Purpura, schoenlein-henoch; Chromatography, high pressure liquid; Mast cells; Histamine release; Shuanghuanglian injection; Spectrometry, mass, electrospray Ionization.