Research Of Anti-cancer Matrine Components In Radix Sophorae Flavescentis And Its Injection Based On HF CF-HPLC

Objective: In the study, hollow fiber cell fishing with high performance liquid chromatography(HFCF-HPLC) and hollow fiber liquid phase microextraction with HPLC(HF-LPME-HPLC) were developed and applied to research active components in Radix sophorae flavescentis(RSF) and its injection in vivo and in vitro, and a dispersive liquid- liquid microextraction(DLLME) was developed to quantify the ir trace level of active ingredients.Methods:HFCF-HPLC based on four living cells(human ovarian cancer SKOV3 cells, human renal tubular ACHN cells, human hepatoma Hep G-2 cells or human breast cancer MCF-7 cells), cell membranes, cell organelles or receptors(cyclooxygenase-2 COX-2, estrogen receptor ER and epidermal growth factor receptor EGFR) was developed and employed to research active matrine alkaloid group(such as oxymatrine, matrine, sophoridine, and oxysophocarpine) from RSF water decoction and its injection in vitro conductting to screen. The matrine components in the joint of the four kinds of cells and the receptor binding ability and the effect of matrine active components and target characteristics were researched. HF-LPME-HPLC based on the blood plasma, urine and the tissue slurry of liver and kidney from rat different means administered in vivo to evaluate and detect the active components which were distributed to organs. Adding organic extract and disperser solvent in turn into the sample solution, based on the characteristics of high enrichment and concentration of DLLME-HPLC, the objective of extracting and determinating of the four trace level matrine alkaloids which the HFCF screened in RSF water decoction and its injection is to comprehend distribution and content of active components(particularly trace level active components) in RSF and its injection.Results: For active components research based on HFCF-HPLC in RSF and its injection:(1) The matrine alkaloids showed potential anti-tumor activity via the four cancer cell lines, membranes and organelles screened by RSF and its injection.(2) The active components of identified oxymatrine, matrine, sophoridine, and oxysophocarpine.(3) The binding ability of the same cell with the same active component in different pharmaceutical dosages showed varying trends, which explained that the binding ability of the active component with the same cell was associated with the state and sample matrix of the active component, and that of the same component with different cells sometimes demonstrated the same trend in different sample matrixes.(4) RSF and its injection exhibited obvious cytotoxicity on the four cancer cells, so both were broad-spectrum antitumor medicines within the scope of the experiment.(5) The main therapeutic mechanism of the active components group screened by HFCF-HPLC may be that the ACHN and Hep G-2 cells could protect against cancer by encouraging apoptosis.(6) The unidentified active components, X1 and X2, which have high target fishing factors(TFFs), may have potential antitumor activity. On the basis of the DLLME condition of optimization, the enrichment factors for four matrine alkaloids ranged from 0.17 to 83.5; the good linearities(sophoridine: 0.0008-6.4 μg·m L-1, oxymatrine: 1.68-606.4 μg·m L-1, matrine: 0.0008-6.4 μg·m L-1, and oxysophocarpine: 0.0008-6.4 μg·m L-1) were obtained for four analytes with regression coefficients of between 0.9924 and 0.9989; the precision(RSD 2.0% to 12.2%), the satisfactory recoveries(92.9-103.2% and 86.6%-104.5%) and the limits of detection were between 0.2-275 ng·m L-1 were also achieved.Conclusion:HFCF-HPLC, coupled with DLLME-HPLC has applied successfully to research trace level matrine alkaloids active components in RSF and its compound injection. Results demonstrated that HFCF-HPLC, coupled with DLLME-HPLC, is a simple, simple, rapid and effective method, which can screen and find the active components(especially the trace active components) in vivo and in vitro, study the entirety and synergy characteristics of multicomponent, multicellular, multiple targets of traditional Chinese medicine. The combination of the three methods provides us novel ideas and novel methods of anticancer efficacy material base and personalized quality control.This study was supported by the National Nature Science Foundation of China(No. 81302734), and the Nature Science Foundation of Shanxi Province, C hina(No. 2011011035-2, 2012011007-4).