The Research Of Purification And Stability Of Reduced Glutathione

Glutathione is an important active substance in body.It's the important agon in many enzyme reactions, which can scavenge free radical and detoxication.This paper introduced the method of the extracting glutathione from the dried yeast.Firstly,use different methods to damage the cell wall o"fan qi"dried yeast,judging from the liquid that contains GSH,we can find"chemical percolating method"is the best one to extract GSH from"an qi"dried yeast,also this method haven't been mentioned in any literature yet.According to the result of my experiments,it is easy to say: the best condition for the experiment should include the following factors:to begin with,put 10 grams dried yeast in a container and then add reasonable quantity of deionized water,mix them until they turn into yeast paste,then use hydrochloric aid buffer to adjust the PH,if it arrive at 3 we can turn to the next step.Further more,heat the yeast paste under the temperature of 60 degrees in a water heated container for 2 hours.Secondly,centrifugal it at a speed of 8000 rounds per minute and the duration is 15 minutes.After all,we collect the remaining liquid of GSH,under the best condition,the quantity of GSH in the liquid(contains 100g dried yeast)would be 72.65mg.Additionally,analyzing"SDS"we can arrive at the conclusion that:there isn't any big numerator in GSH centrifugal liquid.Fortunately,it is good for the purification of GSH later.Then,choose the procedure that"D001"large hole cation–exchange resin palys a role as medium for ion exchanging to separate and extract GSH.It is still a fresh method till now.Meanwhile,I also use"005×7"large hole cation-exchange resin in my experiments in order to gether different data for my futher comparison.After repeating and enlarging experiments I have found the best condition for separating and collecting GSH is as following:frist,buffer liquid:phosphor-acid(pH 4.5,0.02mol/l)contains NACL(10mmol/l) second,the density of added liquid is 100mg/100ml,volume:10ml,lotion:phosphor-acid (PH7.5,0.02mol/l) includesNaCL(0.3mol/l), flowing speed:2.0ml/min.Eventually,the product's color is white.the past experiments have pointed out that the best condition for extracting GSH under the method of"005×007"is that:PH3 ,flowing speed 2.4ml/min,lotion NH4CL(0.05mol/l)but the method is accompanied by low purity and unsatisfactory color of the product.According to my experience we should adopt"HPLC"to test the content of GSH in the product which is not pure.If we use"005×7"to extract GSH,the rate of reclaiming is just 60.3 percentage and the purity is 8.19 percentage.actually, they are 11.3 percentage and 20.71 percentage lower than the result of"D001"respectively.therefore, D001 is the best method to extract GSH.Further more,filtering by gel to get rid of the salt in the product which was extracted from dried yeast under the method of"D001 oin exchanging",Fortunately,the final product's color is extremely white.Exactly,If we adopt"HPLC"to check the content of GSH in the product which is not pure,the average purity is 46.05 percentage,However the reclaiming rate is low.Finally, according to Chinese pharmacopeia (published in 2005) and lots of stable experiments, we can find the best condition for storing GSH is low temperature and dry environment.