I want to study the stability of the folates, including folate, dihydrofolate, tetrahydrofolate,5-methyltetrahydrofolate and 5,10-methylenetetrahydrofolate.Our previous results are the stability for 3,7,14,21,28days.However as I looked for articles about this, I find that it is necessary that we look into the degrading process of the first several hours, because most articles agree that the major reactions happen in the first 24 hours. Contrarily, for our data we can see that sometimes the 3rd day concentration of the folates are even higher than the initial concentration, I believe this is caused by inter-conversions between the folates under different conditions.
Isaac told me that the samples are filtered to get rid of Vitamin D, which means that whatever has a larger molecular weight than Vitamin D will also be eliminated. We checked for the enzymes such as dihydrofolate reductase and methylenetetrahydrofolate reductase, we confirmed that they should be filtered and there should be no enzymatic reactions during the period..
Last time we put all the Vitamin Brsquo;s together and determined their separate concentrations, this time I want to do the folates separately, I want to put them into different tubes and determine the concentration after 1, 2, 4, 8, 16 hours, I expect them to degrade under first order reaction, and I want to see whatrsquo;s the products of the degradation. This experiment could be best done with Lc-Ms, which could both tell me the concentration and the type of molecule.
I concluded from the articles and have my expectations of the products.
5,10-methylenetetrahydrofolate tetrahydrofolate(acidic and neutral conditions,easily)
Dihydrofolate p-aminobenzoyl glutamate and dihydroxanthopterin and 6-formyldihydropterin(pH 5.6,anaerobic oxidized with ferricyanide,room temperature)
Dihydrofolate folate(pH 9.0,anaerobic oxidized with ferricyanide,room temperature)
Folate no breakdown products under pH5.6 and pH9.0,anaerobic oxidation with ferricyanide,room temperature.
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