Honeysuckle Chlorogenic Acid Extraction, Purification And Antimicrobial Properties

Flos Lonicerae is the dried buds or open flowers of Honeysuckle plants, with theefficacy of detoxification, protection of the liver and gall bladder, is the one ofcommonly used herbal medicines. The reason why Flos Lonicerae has those efficaciesis that it contains an active ingredient-chlorogenic acid. Chlorogenic acid is also calledcaffeic acid, molecular formula is C16H18O9, it condensated by the caffeic acid andquinic acid wich are produced by the oxygen breathing of plants, chlorogenic acid hasmultiple biological activities such as antioxidant, anticancer, antibacterial, antiviral, etc.So far many places have developed a variety of products whose main ingredient ishoneysuckle about medicines, health products, functional foods and so on, such ashoneysuckle tablets, herbal tea, toothpaste, etc. However the honeysuckle leaves havebeen discarded as waste, as with the honeysuckle, honeysuckle leaves actually containlarge amounts of chlorogenic acid. So this study choose abandoned honeysuckle leavesas extraction of raw material for chlorogenic acid, and the yield of chlorogenic acid asindex, use the uniform design to research the ultrasonic extraction process and soxhletextraction process, and then compare the two extraction process to determine the betterextraction methods and extraction conditions; by the purification process of chlorogenicacid in honeysuckle leaves, using HPLC and infrared spectroscopy to qualitativelyanalyze the honeysuckle leaves extract; using the D value method to determine the MICof chlorogenic acid on Escherichia coli and Staphylococcus aureus, then researchinginhibitory effection of chlorogenic acid on EC, Aspergillus niger, yeast, SA, Bacillussubtilis. And further investigating the preservative effect of chlorogenic acid in mungbean drink, and get the following conclusions:1The research of chlorogenic acid extraction conditions in honeysuckle leavesThe optimal conditions about uniform design to optimize the Ultrasound extractionof chlorogenic acid in honeysuckle leaves is: the extraction temperature is60℃,extraction time is240min, ethanol concentration is35%, ultrasonic power is200W,solid-liquid ratio is1:40, chlorogenic acid yield is10.5mg/g, and the optimal conditionsabout uniform design to optimize the soxhlet extraction of chlorogenic acid inhoneysuckle leaves is: is the extraction time is240min, ethanol concentration is55%,solid-liquid ratio is1:10, chlorogenic acid was was10.18mg/g. As the result, althoughthe yield of ultrasonic extraction of chlorogenic acid is so closed to the yield of soxhletextraction of chlorogenic acid, the ultrasonic extraction process uses less raw materials than soxhlet extraction process, besides the temperature can be controlled in theultrasonic extraction process.2Purification and qualitative analysis of chlorogenic acid from honeysuckle leavescrude extractBy the complex purification process of metal ion precipitation and organic solventextraction, the chlorogenic acid from honeysuckle leaves crude extract is purified.Using HPLC and infrared spectroscopy to qualitatively analyze the purified extractshows that the purified extract contains Alcoholic hydroxyl, Phenolic hydroxyl,Aromatic ring,-CH2-,-C=C-, ester bond, carboxylic acid, wich consistent with thestructure of chlorogenic acid standard. And after the purification process, the purity ofchlorogenic acid from57.96%increase to81.32%.3The research of antibacterial properties of chlorogenic acid in honeysuckle leavesand its impact conditionsBy D value method and linear regression makes out the MIC of honeysuckle leavesextract on EC is15.05%and on SAis17.57%, then by the filter paper method with18%concentration of honeysuckle leaves extract against EC, SA, yeast, Aspergillus niger,Bacillus subtilis, and these inhibition zone diameter respectively are10.0mm,8.4mm,7.0mm,9.2mm,6.0mm, so honeysuckle leaves extract inhibit tested strains.By filter paper method to initially explore the impact conditions of the inhibitoryeffect of chlorogenic acid, the results show that temperature is the main impactcondition, and the increase of inhibitory effect is by the increasing temperature, butwhen the temperature exceeds60℃, the inhibitory effect will decrease; the pH and UVirradiation time has little effect on its inhibitory effect, but inhibitory effect ofchlorogenic acid in acidic conditions can be slightly better.4Theresearch of preservative effect about chlorogenic acid in mung bean drinkAccording to GB4789.2-2010determine the total number of colonies from mungbean drink where add chlorogenic acid and not add chlorogenic acid after pasteurization,the results show in7℃storage conditions, without adding any preservatives mungbeans drink can be kept only11days, and added1.0g/kg chlorogenic acid mung beandrink can be saved15days, wich indicating that chlorogenic acid certainly can extendthe shelf life of mung bean drink.