Abstract
The rate of hydrolysis of chicken liver folate polyglutamates, by endogenous liver conjugases, under various conditions of storage, heat, and tissue disruption, were investigated. The procedure used was to allow a radioactive tracer dose of the vitamin to equilibrate into the folate polyglutamyl pool. After various storage periods and treatments the polyglutamyl state of the folate present was examined by analytical techniques based on oxidative degradation of native folate polyglutamates to the corresponding p-aminobenzoylpolyglutamate followed by chromatographic separation on DEAE cellulose anion exchange resin. Identification of folate polyglutamates present was made by simultaneous elution of known p-aminobenzoylpolyglutamate markers. In an intact tissue sample only slight degradation was found after 48 hr at 4 C; complete degradation of folate polyglutamates taking 120 hr. Samples of homogenized tissue show complete degradation to folate monoglutamates and a small amount of diglutamate after 48 hr storage. Superimposed on the above is the consideration that if at any time prior to or during storage the liver is heated to greater than 100 C irreversible inactivation of the endogenous conjugases takes place and the folate polyglutamate pattern is stabilized. It was also demonstrated that during two different heating procedures no extra deconjugation occurred.
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