Extraction, Purification And Biological Activities Of Alkaloid From Actinidia Auguta

Actinidia arguta (Siebold et Zucc.) Planch. ex. Miq., known as’kiwiberry’or’baby kiwifruit’ in English, is a perennial, fast growing and deciduous vine. It is mainly distributed in North China, Northeast China, Northwest China, Shandong province, Yangtze river basin and Changbai Mountain in Northeast China. The root, stem, leaf and fruit of A.arguta are valuable with medical and healthy function. Studies demonstrated that A. arguta has extensive bioactivities, such as reducing blood glucose, atopic dermatitis, antioxidant, anti-cancer and anti-allergic.Modern pharmacological studies have reported that alkaloids are the major active ingredients in Chinese herbal medicine and its derived products. Alkaloids containing herbal medicine plants were indicated with activity of anti-inflammatory and antibacterial properties. However, little attention has been devoted to function of alkaloids extracted from A. arguta.The objective of this research is to accelerate the usage of A. arguta as an important functional food, a series of studies are made and the main results are as follows:1. The results show that alkaloids exist in root, stem, leaf and fruit of A.arguta with the study of vitro sedimentation.2. The measuring method of A. arguta alkaloids are found firstly, as follows:aconitine as standard, the optimal buffer pH is 7.5 and amount is 6 mL; the optimal dosage of bromothymol blue is 2 mL; wavelength scanning is performed at 415 nm. The results show that the most alkaloids consist in root and the least in leaf.3. The optimum technological conditions is found by single factor and orthogonal experiment, as follows:the optimal concentration of ethanol is 60%; the optimal solid-liquid ratio is 1:25 g/mL; the optimal ultrasonic power is 200 W and 15 min. The extraction ratio of alkaloids is 0.683 mg/g.4. D-101 is the optimum macroporous resin. The optimum technological conditions of adsorption as follows:the optimal concentration of alkaloids is 0.4 mg/mL; the optimal pH is 10; the optimal flow rate is 2 mL/min; the optimal fluid volume is 3 BV; 3 BV water flushing of the resin column; 3.4 BV,80% ethanol solution at the same rate of elution. The results show that the average recovery of A.arguta alkaloids is 90.9%; the average purity is 77.6%.5. The composing component of A.arguta alkaloids is first identified with HPLC and LC-MS. The results show that the main ingredients of A.arguta alkaloid are aconitine, berberine, yanhusuo alkali, tetrahydropalmatine, aconitum original alkali, lentils, atropine, actinidine.6. The antioxidation in vitro activities of A.arguta alkaloids are first identified. Compared with Vc, the alkaloids of A.arguta have strong DPPH radical scavenging and resistance to liposome peroxidation activities; It has good reduction activity to Fe3+, but is weaker than Vc; It can clear OH· and O2-· to a certain extent but the activities is weaker than Vc obviously. The results show that A.arguta alkaloids has antioxidant activity in vitro to a certain extent.7. The immune function in vivo by the A. arguta crude alkaloids (AACA) is first studied. The results show that organ coefficients including liver, spleen and thymus coefficients were improved by AACA treatment. But the difference was not significantly with spleen coefficient. AACA could improve the weight of liver and spleen in mice, and then increase corresponding carbon clearance index. The treatment of AACA effectively increased the levels of IL-6, IFN-γ and LSZ in normal and immunosuppressed mice. Furthermore, AACA could enhance the antioxidant enzyme activity of SOD, CAT and GSH-Px in liver. The results show that AACA could improve the immunity of mice.8. The activities of the anti-fatigue and increasing exercise performance effect of AACA are first found. The results show that AACA could increase significantly the time of weight-loaded swimming test and forelimb grip strength in mice with AACA-100 group compared to the vehicle. Further, AACA elevate exercise performance by increasing levels of glucose, albumin, TP and decreasing levels of lactate, ammonia, CK, UA, BUN. The reserves of hepatic and muscular glycogen have been increased after exercise, which could provide energy for the body movement and improve exercise performance. It provided evidences to support that alkaloids extracted from A.arguta as a novel anti-fatigue and exercise performance agent with physiological benefits.