Preparation Of Silybin Microparticles And Their Association Rules By Stainless Steel Membrane Emulsification

Background:Apply stainless steel membrane emulsification method to prepare silybin micro-particles with uniform and controlled size and they had no excipients. Then optimize its preparation by the uniform design and orthogonal design. In order to establish association rule model of microspheres and micro-particles, research on physicochemical properties of silybin PLGA microspheres and micro-particles without excipients had been conducted. Investigate targeting bioactivity in mice to provide methodological support for more effective components of traditional Chinese medicine in micro-particle delivery system preparation. Methods:On the basis of no supplement agents and apply rapid membrane emulsification coupling solvent evaporation method to prepare silybin micro-particles. Take the average size (D (0.5)) and distance (span) as the index, optimized preparation process by uniform design and orthogonal design method using stainless steel membrane tubes with size of 0.5μm of multi-stage series; the physicochemical characteristics of these microspheres and particles were determined by scanning electron microscopy, differential scanning calorimetry, and powder X-ray diffraction; association rule data mining model was established to investigate correlation law among process parameters of silybin PLGA microspheres and particles preparation, and reveal the main influence factors and levels of qualify of silybin microspheres and micro-particles; the targeting bioactivity in Kunming mice were assessed by giving tail vein injection of silybin micro-particle suspension, in order to access microscopic distribution and targeting mechanism of silybin micro-particles in vivo, blood and liver were taken, then observe frozen liver slices by scanning electron microscopy, further, stimulate and isolate peritoneal macrophages in mice after I.P., finally observe phagocytosis of macrophage. Results:Study showed that the optimum conditions were as follows:membrane pore was size 0.5m (4-stage series connection), ratio of oil and water phase volume was 2:3, oil phase concentration was 0.8%, PVA concentration of was 3%, kind of curing liquid was 3% PVA, membrane pressure was 0.6Mpa, curing time was 60min. Prepared microspheres were uniform and with a smooth surface, the mean diameter was (2.93±0.32) μm, the span was (1.01±0.04). Moreover, dispersibility was well, drug was molecular or amorphous in silybin PLGA microspheres, unsealed drug was still fine crystal, while in silybin micro-particles drug existed as crystal. Further, the correlation laws by the date mining as follows:agitating magnetically, the stainless steel tube and the concentration of PVA were less than or equal to 3%, the above conditions were qualified. In addition, silybin micro-particles was visible in the liver sinus lenticels, when the frozen liver slices were observed by scanning electron microscope, in particular, the trace of silybin was detected in peritoneal macrophages, mesentery and blood cells. Conclusions:Silybin micro-particles were assessed in the physicochemical properties, association rule model and other aspects. The micro-particles prepared using the optimal method had a uniform-sized, controlled particle size and stable quality. Stainless steel premix membrane emulsification apparatus was feasible in the micro-particles preparation. Silybin micro-particles distributed with systemic blood through the body after injected into mice tail vein. The free bound plasma proteins, part were engulfed by mononuclear cell, perhaps part were retained in liver Disses’space and lenticels, part were engulfed by kupffer cells, part particles with slow blood accumulated in the terminal capillary network rich in blood (such as the mesentery) etc..