| Objective:To investigate the extraction, purification and antitumor effect of total flavonoids from Valeriana jatamansi Jones.Method:Taking total flavonoids as indexes, ultrasonic technology was applied to extract, and the optimal extracting parameter was worked out by univariate analysis of variance and orthogonal test. The purification of total flavonoids was studied after extracting, the static absorption-desorption was used and compared in order to find the optimum macroporous resin, and the technical process for purification of total flavones with the optimum macroporous resin was screened by univariate analysis experiment, with yield of total flavones as index. H22 and S180 tumor bearing mice models were established to study the antitumor effect of total flavonoids from Valeriana jatamansi Jones.Results:Taking total flavonoids as indexes, the optimal extracting parameter is as follows: ethanol concentration was 65%, ratio of solid and liquid was 1:20, ultrasound time was 20 minutes and ultrasound times was 3.The HPD600 macroporous resin among five types of resins was found with the best separating efficiency, by which static absorption capacity and desorption ratio could reach 131.11 mg·g-1 and 91.69%, respectively. The best result of dynamic absorption was based on the followings:sample concentration-3.42 mg·mL-1, feed rate-2.4BV·h-1, sample capacity-25.64mg·g-1, elution flow rate-2.4BV·h-1. After eluted with 3BV of distilled water and 4BV of 80% ethanol, the yield of total flavones was 80.36% and the purity was 41.20%.The total flavonoids of Valeriana jatamansi Jones had an evident antitumor effect in vivo. The inhibition rates on H22 tumor were 19.25%,30.06%,25.97% and 45.32%, respectively in the concentration of 450 mg·kg-1,150 mg·kg-1,50 mg·kg-1 total flavonoids of Valeriana jatamansi Jones and active-controlled drug. Compared with average tumor weight of model group,150 mg·kg-1,50 mg·kg-1 total flavonoids of Valeriana jatamansi Jones and active-controlled drug have significant difference(p<0.05,p<0.05,p<0.01).The inhibition rates on S180 tumor were 30.80%,41.53%,45.57% and 59.97% respectively in the concentration of 450 mg·kg-1,150 mg·kg-1,50 mg·kg-1 total flavonoids of Valeriana jatamansi Jones and active-controlled drug. Compared with average tumor weight of model group,150 mg·kg-1,50 mg·kg-1 total flavonoids of Valeriana jatamansi Jones and active-controlled drug have significant difference(p<0.05, p<0.05, p<0.01). The observation on immune organ of H22 and S180 tumor-bearing mice indicated that compared by thymus index of normal group, H22 and S180 model groups had downward trends but no difference, active-controlled group had downward trend and significant difference(p<0.01), and three total flavonoids groups of Valeriana jatamansi Jones showed increasing trends, and 50 mg·kg-1,50 mg·kg-1 total flavonoids of Valeriana jatamansi Jones had significant difference(p<0.05). The pathomorphological observation indicated that the tumour cell of H22 and S180 tumor-bearing mice were more and close, their cellular nucleus were big, presenting obvious heteromorphism,while total flavonoids groups of Valeriana jatamansi Jones and active-controlled group appeared cell necrosis in different degree.Conclusions:By univariate analysis of variance and orthogonal test, the optimal extracting parameter of total flavonoids of Valeriana jatamansi Jones was:ethanol concentration was 65%, ratio of solid and liquid was 1:20,ultrasound time was 20 minutes and ultrasound times was 3. By univariate experiment, the optimal purification parameter of total flavonoids of Valeriana jatamansi Jones was:HPD600 macroporous resin, sample concentration-3.42 mg·mL-1 feed rate-2.4BV·h-1, sample capacity-25.64mg·g-1,elution flow rate-2.4 BV·h-1. After eluted with 3BV of distilled water and 4 BV of 80% ethanol, and the purified total flavonoids of Valeriana jatamansi Jones have antitumor effect, and the two concentrations of 150 mg·kg-1,50 mg·kg-1 had better effect. |
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