Research On The Anti-fatigue Effect And Product Development Of Ruditapes Philippinarum And Its Compound

Because of intensive modern living style,more and more people are easy to feel tired,irritable,fatigue.However,there is few related health food in the market.Not only were anti-fatigue effect of Ruditapes philippinarum and Polygonatum Lilium Meretrix Decoction(PLMD)researched according to old literatures and modern research reports but also technological processingand toxicology research of PLMD.In the system of traditional medicine theory,Ruditapes philippinarum is beneficial to help five zang-organs running,according to Jia You Materia Medica.In the previous research,our research group found that among four low value shellfish,Mactra veneriformis,Ruditapes philippinarum,Cyclina sinensis and Meretrix meretrix in the coastal of Jiang Su Province,Ruditapes philippinarum has the highest content in taurine,essential amino acid and nucleosides,besides,polysaccharide and protein components are rich.Among them,taurine has a clear anti-fatigue effect,some amino acids and polysaccharides,protein components have a certain anti-fatigue effect.Based on the theories of traditional Chinese medicine and modern scientific research,the an-fatigue effect of Ruditapes philippinarum and PLMD were evaluated to develop a safe and effective anti-fatigue health food with Ruditapes philippinarum as raw material.This paper mainly includes the following parts:1.Literature studyAfter referring to the relevant research materials of fatigue and anti-fatigue activity substances both domestic and foreign,learn the diverse concepts,treatment principles and methods between traditional Chinese medicine and Western medicine to lay the solid foundation for the future research work.2.Study on the anti-fatigue effect of Ruditapes philippinarumReferring to Technical Standards for Testing and Assessment of Health Food(2003)and the related literature,mice burden swimming fatigue model were conducted,time of weight-loaded swimming,level of LA and LDH.CK in serum after swimming without load to evaluate the anti-fatigue activity of Ruditapes philippinarum.Consequently,it was found that extractives of Ruditapes philippinarum could prolong the swimming time and decrease the content of lactic acid in serum,and decrease the activity of LDH and CK.Ruditapes philippinarum extraction has a certain anti-fatigue effect.3.Study on acute toxicity and anti fatigue effects of Polygonatum Lilium Meretrix DecoctionPolygonatum Lilium Meretrix Decoction was found to solve the problems of fishy smell and large dosage of Ruditapes philippinarum.Subsequently,the anti-fatigue effect and acute toxicity test of PLMD were evaluated in mice.Compared with Ruditapes philippinarum extraction,PLMD’s anti-fatigue effect was better.PLMD could improve not only the forced swimming time,levels of LA,LDH,CK,but also SOD and MDA.Maximum tolerance test result showed that PLMD’s MTD is 342 g/kg(crude drug),it is safe and nontoxic.4.Study on the process of decoction and desalination of Polygonatum Lilium Meretrix DecoctionLily polysaccharide has been confirmed with anti-fatigue effect,while taurine has anti-fatigue effect clearly.Therefore,polysaccharides and taurine were choosed as the functional composition.Anthrone sulfuric acid method and OPA pre-column derivatization HPLC method were used to determine polysaccharides and taurine,respectively.The UV wavelength scanning result indicated the wavelength of 580 nm was the best.The methodology study showed that this method has good linearity.Pecision,repeatability and stability test(RSD)values were 1.25%,2.36%and 2.13%.The rate of average recovery was(97.85%+0.52)%,fulfilled the requirements of sample analysis.The detection of taurine was by OPA pre-column derivatization HPLC method,which was detected in the wavelength of 330 nm at a flow rate of 1 mL/min,the column temperature at 40℃,VMethanol:V0.05mol/L sodium acetate=40:60,in 10μL of sample volume.It was proved that this method with good specificity,the linear relationship was good when taurine was in 0.01003～1.00280 mg/mL concentration range.The regression equation was Y=2E+06x-865,R 2=0.9999.As for the precision,repetition,stability,and the rate of recovery were conformed to the requirements of sample analysis.The preparation condition of PLMD was optimized by the single factor and orthogonal design with extractive yield amount of polysaccharides and taurine as index.The best decoction technique was adding 10 fold of water decocting three times 60 minutes each time.Nevertheless,ash content and conductivity of PLMD were high because of Na+,K+ and other inorganic ions and heavy metal ions,it has a negative effect of PLMD’s taste and health.Electrodialysis device was used to desalt PLMD with desalination,the transfer rate of yield,total sugar and taurine as the evaluation indexs.The optimum extraction of desalination was desalting for 90min,the sample loading quantities is 50%of the capacity of electrodialysis device and the voltage of desalination is 1.0V/on the membrane.5.Study on anti-fatigue and acute toxicity of Polygonatum Lilium Meretrix Decoction after desalinationForced swimming test was conducted to evaluate the anti-fatigue function of desalted PLMD.The results show that high dose group of desalted PLMD could significantly prolong the swimming time(P<0.05),LA(P<0.01),LDH(P<0.05)and CK(P<0.05)levels have decreased significantly,at the same time,the activity of SOD increased and the content of MDA declined.Anti-fatigue function of PLMD after desalination was still remained.The MTD of desalted PLMD is 368.4 g/kg(crude drug),it has changed little compared with non-desalted.6.The preliminary syudy of Polygonatum Lilium Meretrix Decoction GranulationThe effective dose of desalted PLMD is large,so granule was selected to be researched.Spray-drying method was adopted to prepare PLMD powder,and then added appropriate amount of adhesive,wet granulation,whole,dried,be finished.Detection method of taurine and polysaccharide was established to control product quality and provide a better method for further research.