Study On The Technology Of Extraction And Separation And Of Cichoric Acid Physiological Activity From Cichorium Intybus L.

Cichorium intybus L. has significant health promoting effects and great application value,and has been used for the production of food and drug industry, especially as an agent to enhance immune system function, antioxidant and anti-virus because of cichoric acid.Cichoric acid composed of caffeic acid and tartaric acid belongs to caffeic acid derivatives,which is one of the most abundant phenolic compounds and immune active ingredients. In this study, the following experiments were conducted successively on the stems of Cichorium intybus L. Extraction and purification parameter from Cichorium intybus L. were investigated,at the same time, antioxidative and Tyrosinase inhibition properties were tested. The specific findings were as follows:1. Single factor experiments and the orthogonal experiment design methods L9(34) were applied to analyze the influencing factors through the ultrasonic-assisted extraction. The best process parameters were as follows: 50% ethanol, 60℃, 50 min, solid-liquid ratio 1:21, 180 W of ultrasonic power, the optimum yield of cichoric acid was 1.51mg/g under this condition.Meanwhile, some by-products were obtained as follows: tartaric acid 0.34 mg/g, chlorogenic acid 0.55 mg/g, caffeic acid 0.13 mg/g, galuteolin 0.80 mg/g. Temperature, solid-liquid ratio and ultrasonic power had a significant effect on cichoric acid yield(P<0.05).It can improve the yield of cichoric acid compared with the traditional process, which are characteristics of high efficiency economy.2. Using of reverse osmosis equipment concentrated cichoric acid extracts, the optimum concentration condition were as follows: Operating temperature was 50℃, Operating pressure was 20.0 bar, the average water flux of membranes was 41.0 L /m2/min under this condition.The cichoric acid extracts was concentrated to the original volume of 15%. Cichoric acid was purified by using N-butyl alcohol extraction method, and extraction of temperature, time and the amount of solvent was test on its conctration. The optimum process conditions were determined as follows: Extraction temperature 50℃, time 40 min, liquid ratio was 1:2, the optimum yield of cichoric acid reached 0.41 mg/mL. At last, in order to make the cichoric acid and the other components isolate and purify, the preparative liquid chromatography wasutilized. The mobile phase were methanol-0.1% formic acid water, flow rate was 7.0 m L/min.With eventually mass spectrometer to identify cichoric acid and the other components.Cichoric acid purity was reached 93%, and other by-products, purity were as follows: tartaric acid 91.5%, caffeic acid 95%, galuteolin 98%, respectively.3. There was a concentration-response relationship between cichoric acid extracts concentration and antioxidant activity. IC50 values against ·DPPH, ·OH free radical and ferric reducing antioxidant power were 4.97 mg/L, 5.73 mg/L and 5.61 mg/L, respectively. The ability of scavenging ·DPPH and ·OH free radical was as follows: cichoric acid > galuteolin >chlorogenic acid > caffeic acid > caftaric acid, and the ability of ferric reducing antioxidant power was: caftaric acid > caffeic acid > galuteolin > chlorogenic acid >cichoric acid. There were synergistic antioxidant effect among the three antioxidants, and galuteolin was stronger than cichoric acid and chlorogenic acid.4. Cichoric acid can efficiently inhibit both monophenolase and diphenolase activities of tyrosinase. 0.40 mmol/L cichoric acid can extend the lag time from 133 s to 383 s in the oxidation process of L-tyrosine via tyrosinase, the time was relatively increased 2.87 times,and the inhibitory rate reached 84.3%. The cichoric acid concentrations corresponding to 50%inhibitory rate on monophenolase and diphenolase activity were 0.26 mmol/L and0.71mmol/L, which were much lower than arbutin for diphenolase activity( IC50=5.30mmol/L). Lineweaver-Burk plot demonstrates a reversible competitive behavior of cichoric acid in the tyrosinase oxidation of L-DOPA, with maximum reaction rate vm and inhibition constant KI at 83.30 μmol/L and 0.14 mmol/L, respectively.