Extracting Process Optimization Of Iridoids From Valeriana And Their Antioxidation In Vitro

Valeriana belongs to Valeriana L. in Valerianaceae. It has significantly effects on:smooth muscle antispasmodic, increase coronary blood flow, antiarrhythmic, anti-tumor, tranquilizing and so on. Currently, the study of effectiveness and ehemical composition analysis bout Valeriana is popular, but the study of process optimization rarely. The natural active substances of iridoids from the V. officinalis var. latifolia was extracted by the ultrasonic extraction, separated by the macroporous resin purification and silica gel column chromatography. We design experiments for discussing antioxidation in vitro of iridoids purified by macroporous resin from V. officinalis var. latifolia. Especially the process conditions. The results of study were as follows:(1) The optimum conditions of ultrasonic extraction of iridoids form V. officinalis var. latifolia Through the quadratic general rotary unitized design, ultrasonic extraction mathematical simulation model was concluded by the linear regression analysis:Y=1.63814+0.03575X1-0.02958X2-0.00958X3+0.02350X4-0.03433X12-0.04620X22-0.03920X32-0.03295X42-0.01000X1X3+0.01187X2X3 Then the optimum conditions of ultrasonic extract iridoids form V. officinalis var. latifolia was determinded by the mathematical statistical analysis:ethanol concentration is 80%, ultrasonic power is 250 w, extract time is 5.5 min, ratio of material to liquid is 22:1. Its extraction rate on theory which was calculated by mathematical simulation model can be achieved 1.66%. The theoretical model was proved that was reliable by the verification experiments, the value is 1.66%.(2) The process optimization of iridoids purified by macroporous resin After the type optimized test of macroporous adoption resin, D101 resin the loading volume test and the dynamic eluting test and the column chromatography quadratic general rotary experiments on three factors (loading velocity, eluting velocity and concen-tration of eluting solvent). the optimum conditions of D101 macroporous resin column chromatography test was determined by these experiments and all kinds of analysis: the concentration of sample is 10.0 mg/mL, pH=6.8, the upper sample volume is 20 mL, elution volume is 4 VB, macroporous resin purification mathematical model of iridoids can be achieved:Y=70.06392-3.69212X1- 1.84568X2+6.43739X3-2.67725X12-3.16338X22-7.88863X32+2.35000X1X3-1.57000X2X3, Through regression analysis and variance analysis, the optimum conditions on macroporous resin purification were drawn:the sample velocity is 2.5 BV/h, the elution flow is 1.7 BV/h, the ethanol concentration is 75%, the yield rate of purified iridoids is 72.40%, which was measured by HPLC, and the purity of separated iridoids can reach 46.70%.(3) The purification of iridoids by silica gel column chromatography Through the TLC, silica gel column chromatography and HPLC purity experiments, chloroform and methanol in the ratio of 7:1 as the eluting solvent, used the ratio can be achieve good separated effect to target product. With the silica gel column chromatography the purity of purified iridoids powder by the macroporous adoption resin can be advanced from 46.70% to 98%. The purificated products were valtrale Judged form characteristics reaction and HPLC detect technology.(4) Antioxidantion in vitro research of the iridoids purificated by macroporous resin In the experiment of scavenging on free radical invitro, the purification of iridoids showed the scavenging effect on DPPH, hydroxyl radical(-OH), superoxide anion radical (O2·) and the scavenging order was DPPH>O2·>·OH, the half scaven-ging concentration (IC50) were 1.33,1.45,2.07μg/mL, respectively.