Study On Optimizization Of Extraction Process And Antifungal Mechanism From Ginnamon

The Cinnamon extracts had best inhibiting effect on Candida albicans screened from alcoholic extracts of 24 spices. And its major antibacterial compositions were preliminarily analyzed by thin layer chromatography-bioautography and high performance liquid chromatography. Based on the single factor experiment and the Box-Benhenken response surface analysis as well as response surface test and regression analysis, Cinnamon ethanol extraction process was optimized. The best process of Cinnamon alcohol extraction was as follows:extraction temperature is 69.24℃, the liquid-solid ratio is 10:1 (mL:g), extracting time is 5.15 h, ethanol concentration is 95%. Under the optimal conditions, the antibacterial circle against Candida Albicans is 39.46 mm. While developing agent was petroleum ether:ethyl acetate= 9:1 (v/v), Compared with Cinnamon extracts and Cinnamyl aldehyde, Cinnamyl aldehyde was initially identified the main inhibitory ingredient in the Cinnamon. The result of high performance liquid chromatography showed that the major antibacterial composition of Cinnamon extracts was Cinnamyl aldehyde.Antibacterial activity to Candida albicans, such as proteins released, SDS-PAGE, alkaline phosphatase (AKP) content, lactate dehydrogenase(LDH) released, the measurement of electrical conductivity, K+and inorganic phosphorus content, and the changes of UV absorption material and succinate dehydrogenase (SDH) were detected. Those initially reseached the antibacterial mechanism of Cinnamon aldehyde of Cinnamon extract on Candida albicans. The results indicated that Cinnamon aldehyde of Cinnamon extract could damage the structure of cell wall and cell membrane, which resulted in the increase of permeability of cell membrane and release of cell components. Cinnamon aldehyde of Cinnamon extract could also affect the synthesis of Candida albicans’protein, blocking the protein expression in bacteria.The antimicrobial mechanism of Cinnamon aldehyde of Cinnamon extract against Candida albicans was assayed by flow cytometry and electron microscope. The minimum inhibitory concentration of the extract against Candida albicans was 3.125 mg/ml. Then Candida albicans were disposed by different concentration of extract:1 MIC,2 MIC and 4 MIC, and the change of FSC, DNA, membrane potential, cell cycle, Ca2+and Annexin V-FITC/PI double staining were analyzed by flow cytometry and cell morphology were studied by transmission electron microscope and scanning electron microscope. The results showed that the extract inhibited Candida albicans by destroying of cell membrane system and blocking DNA.