Evaluation Of Anti-fatigue Functional Factors And Development Of Walnut Functional Drinks

In recent years,fatigue has become an important factor endangering human health and has aroused widespread concern.Screening safe and effective anti-fatigue active ingredients,designing and developing anti-fatigue products are in line with current market needs,and are of great significance to human health.This project using walnut milk as the base material,through the screening of anti-fatigue functional factors and the evaluation of anti-fatigue,aims to develop a new walnut functional beverage to prevent fatigue,which is tested by combining internal and external body.To evaluate and evaluate anti-fatigue effects,anti-oxidation is used as the basis for judging anti-fatigue effects in vitro.Animal fatigue models are established through exercise tests in vivo,and the corresponding physiological and biochemical indicators are measured,in order to obtain a walnut functional drink with good anti-fatigue effect.The main research contents and results are as follows:First of all,the single-factor,orthogonal and response surface tests were used to optimize the preparation process formula of the basic walnut milk and the compound stabilizer formula.Taking the sensory evaluation of walnut milk as the index,using the material-liquid ratio,the amount of sucrose added,the refining time,and the homogenization time as the experimental factors,the optimal preparation process and formula of walnut milk were optimized by using single factors and orthogonal experiments:The ratio is 1:20,the sucrose content is 6%,the refining time is 20min,and the homogenization time is 15min.Under this test condition,the walnut milk tastes smooth;the sweetness is moderate;and the system is stable.Next,the effects of different amounts of emulsifiers and stabilizers on the average particle size,emulsification stability and precipitation rate of walnut milk were studied.The walnut milk complex stabilizer was obtained through response surface optimization experiments based on the comprehensive score of walnut milk stability.The formula is:the added amount of monoglyceride 0.12%,the added amount of sucrose ester 0.15%,the added amount of xanthan gum 0.05%,the added amount of sodium alginate 0.05%.The average particle size of walnut milk prepared with the composite stabilizer is small;the overall level of emulsion stability;precipitation rate is low;and the system is stable.Secondly,through vitro antioxidant tests,we screened different molecular weight walnut peptides,walnut proteins,and five exogenous functional factors.Studies have been made to extract walnut protein from walnut meal by alkali-soluble acid precipitation method;walnut peptides were prepared by enzymatic hydrolysis,walnut peptides of different molecular weights were separated by dialysis bags,and amino acid composition was determined by HPLC method.Based on the measurement of ABTS⁺·,DPPH·,·OH scavenging ability and reducing power,the in vitro antioxidant capacity of various functional factors was evaluated comprehensively.The results showed that walnut peptides of different molecular weights contained 18 amino acids,but the contents of each amino acid were not the same.Among them,aspartic acid,alanine,glutamic acid,and arginine were the main constituent amino acids.Walnut peptides of different molecular weights and walnuts proteins both have good anti-oxidant effects and are dose-dependent.Among them,walnut peptides of 0 to 1000 Da have the strongest antioxidant capacity.The IC50 values for clearing ABTS⁺·,DPPH·,·OH and reducing power were 0.632mg/ml,0.340mg/ml,0.490mg/ml,3.51mg/ml.The five exogenous functional factors all have good antioxidant capacity in vitro.Among them,tea polyphenols are the strongest,followed by taurine.The IC50 values for the removal of ABTS⁺·,DPPH·,·OH and reducing power are0.094mg/ml and 0.290mg/ml,0.251mg/ml and 0.684mg/ml,0.393mg/ml and 1.142mg/ml,0.838 mg/ml and 0.652mg/ml.Thirdly,by constructing a mouse exercise fatigue model,an open field experiment,and measuring physiological and biochemical indexes in serum,skeletal muscle and liver,a variety of functional factors were evaluated for fatigue resistance.The study evaluated the effects of various functional factors on exercise tolerance and fatigue performance of mice through exercise experiments such as pole climbing experiments,suspension experiments,swimming experiments,weight-bearing swimming experiments and open field experiments;the anti-fatigue effect of various functional factors was evaluated by measuring the changes of physiological and biochemical indexes of lactic acid,liver glycogen,muscle glycogen,urea nitrogen in mouse serum,liver or skeletal muscle;by measuring glutathione peroxidase activity?GSH-PX?,Catalase activity?CAT?,superoxide dismutase activity?SOD?and malondialdehyde?MDA?contents were analyzed for the antioxidant activity of various functional factors in vivo.The results show that:walnut peptides,walnut protein,soy peptides and five exogenous anti-fatigue factors can improve exercise endurance in mice to some certain extent,and have certain anti-fatigue effects.Among them,walnut peptides,L-carnitine and the effect of taurine are more significant.The results of empty field experiments show that several anti-fatigue factors can improve the ability of mice to explore unfamiliar environments,autonomy and adaptability to some certain extent,relieve anxiety and nervousness.Among them,walnut peptides,L-carnitine and taurine,the effect of acid is more significant.The results of fatigue physiological and biochemical indicators show that several anti-fatigue factors can enhance the aerobic metabolism of lactic acid,reduce the lactic acid content in serum and skeletal muscle,increase the content of liver glycogen and muscle glycogen,and effectively clear the accumulation of serum urea nitrogen.Among them,the effect of walnut peptide,L-carnitine and taurine are more significant,and the anti-fatigue effect are better.Antioxidant physiological and biochemical indicators showed that walnut protein,walnut peptides,and soy peptides can all reduce the MDA content in serum and organs of mice after vigorous exercise,accelerate the timely removal of MDA,and avoid oxidative damage in the body.The serum,skeletal muscle and liver WEP,WPP,SP groups have stronger GSH-PX enzyme activity,CAT enzyme activity and SOD enzyme activity than the NC group,which can effectively remove active oxygen free radicals from the body,of which WPP The group had the strongest antioxidant enzyme activity,the strongest total antioxidant capacity,and the best antioxidant effect.Finally,the screened three functional factors of taurine,L-carnitine and 0¹000Da walnut peptide with good anti-oxidation and anti-fatigue effects were formulated.The preparation process of anti-fatigue walnut functional beverage was optimized by single factor and response surface test.The experimental results show that when the taurine:L-carnitine:walnut peptide is 1:1:2,the clear effect on DPPH free radicals is the best,with an IC₅₀0 of 38.68?g/mL.Through single factor and response surface experiment screening,the preparation process of taurine,L-carnitine and walnut peptide composite anti-fatigue walnut milk was optimized,and the formula of anti-fatigue walnut milk was obtained:the material-liquid ratio is 1:20,the amount of sucrose is 6%,the added amount of the compound stabilizer is 0.3%,and the added amount of the anti-fatigue factor is 6g/L.The walnut milk prepared under this condition has a delicate taste,rich aroma and good anti-fatigue effect.