Studies On Extraction Process Of Chlorogenic Acid From Flos Lonicerae

AIM:To establish a method to determine chlorogenic acid from Flos lonicerae and Jinqi Jiangtang Tablet.Optimize the extraction and purification process of chlorogenic acid from Flos lonicerae and its effect on a-amylase had been studied in this thesis.METHODS:A method for determining chlorogenic acid from Flos lonicerae and Jinqi Jiangtang Tablet had been established by using reversed-phase high performance liquid chromatography (RP-HPLC). In the study on Flos lonicerae extraction technology, solvent extraction and ethanol extraction were investigated. Then the Flos lonicerae was treated by means of engyme before extracted by water, the effect of relative factors studied by HPLC, such as enzyme dosage、treatment time、 pH and treatment temperature of cellulose,pectase and mixed enaymes on extraction yield were discussed, and then designing orthogonal test according L9(34)factor levels for optimuming technology of extracting Flos lonicerae. Studying semi-bionic extraction of chlorogenic acid from Flos lonicerae by the same way. Comparison of purification methods of chlorogenic acid with ethanol and ethyl acetate in detail.To study influence of chlorogenic acid from Flos lonicerae on a-amylaseRESULTS:To use RP-HPLC for determination. The separation was carried out using the Diamosil C18column (4.6mm×250mm,5μm) with the mobile phase containing acetonitrile-0.4%orthophosphoric acid (13:87) and a detective wavelength at327nm. The flow rate was1.0mL·min-1. The method was simple, accurate with a good recovery and repeatability. The method can be used to control the quality of Flos lonicerae and Jinqi Jiangtang Tablet. Water as the optimal extraction solvent,and the extraction conditions obtained were following:extracted the chlorogenic acid from Flos lonicerae2times and1h each time at70℃,the first time1:15(W/V) of water and the second time1:10(W/V) of water. Through the factorial and the orthogonal design,the optimum extraction process consists of adding the cellulose by0.5%, pH6.0, the optimum temperature was40℃and enzymatic hydrolysis for1.5hour with the chlorogenic acid was2.38%, adding the cellulose by1.0%, pH4.0, the optimum temperature was50℃and enzymatic hydrolysis for1.5hour with the chlorogenic acid was2.25%, adding the mixd enaymes(cellulose:cellulose-1:1) by0.2%, pH5.5, the optimum temperature was45 ℃and enzymatic hydrolysis for1.5hour with the chlorogenic acid was2.46%. The optimal semi-bionic extraction conditions obtained was adding the solvent adjusting by sulfuric acid and sodium hydroxide which pH were2.2and7.4respectively and2h each time,1:20(W/V) solvent, the optimum temperature was70℃with the chlorogenic acid was2.14%. Ethanol with concentration of75%for12h was optimal purification methods after inspissation. It was found that chlorogenic acid could inhibit the activity of a-amylase in this thesis. The inhibiting ratio of chlorogenic acid on a-amylase was96.22%.CONCLUSION:The confirmed extraction and purification process of chlorogenic acid from Flos lonicerae which shows high yield rate of active constituents and good reproducibility.The method of determination which is simple, accurate with a good recovery and repeatabilityand can be used to control the quality of Flos lonicerae and Jinqi Jiangtang Tablet. It is found that chlorogenic acid could inhibit the activity of a-amylase,and the potential methanism may be related to inhibiting the a-amylase activity and delaying the intestinal absorption of carbohydrates.