Enzymatic Extraction And Anti-aging Effect Of The Trapa Bispinosa Roxb Extractive

The purpose of this study is to improve the technology for extraction of Trapa bispinosa Roxb (TbR) shell,and to investigate the anti-free-radical ( anti-aging ) effects on this shell extracts. The results may provide a foundation for further pharmaceutical use of TbR.Using the extraction rates as the evaluating criteria, extraction techniques were studied with or without enzyme, using cellulase or pectinase, by varying different ratio of two enzymes. Temperature, extraction time, amount of enzymes, ratio of two enzymes were the factors considered in a L₉(3⁴) orthogonal design. An optimized procedure with temperature at 50℃, reaction time at 60 minutes, amount of enzyme used (weight ratio of enzyme and TbR shell) at 1.8%. enzymatic ratio ( weight ratio of cellulase and pectinase) at 2 to 1 was selected.The new procedure provided an extraction rate of 0.175%, a rate that is three times higher than the classical extracting method. Protein purification by type II ZTC1+1 natural clarifying agent reduced the protein content of the new extracting product from 12.43% to 6.33%.The pharmacological evaluation on anti-aging effects of TbR shell extracts obtained by enzymatic and classical methods, and before or after protein purification, were carried out on aging mice model produced by sub-dermal injection with 100mg/kg of D-galactose on animal's dorsal side of neck-back area for 35 days. The results showed no difference between the two extracting methods. However, deproteinized products was found to have lower anti-oxidative capacities.Anti-aging effects of three levels of testing drug TbR shell extracts by enzymatic method) were evaluated on aging mice by measuring serum and liver levels of T-SOD, GSH-Px and MDA, as well as the liver levels of MAO-B and lipofuscin. Vitamin E was used as the positive control drug. SPSS10.0 was used in statistical analysis. The results indicated that all the three levels of testing drug were effective antioxidents, and there was no difference on their pharmacological anti-aging effects. When comparing the high testing drug group with vitamin E group, the serum and liver levels of T-SOD and GSH-Px were higher (p<0.05) while MDA was lower (p<0.05) in the former. No difference was found for liver concentrations of MAO-B and lipofuscin when the above two groups were compared.The results of this study showed that TbR shell extracts enhanced the anti-oxidative capacities and reduced the formation of lipid peroxides in the aging mice model. Therefore, TbR shell extracts have the potentials to be further developed into an anti-aging pharmaceutical agent.