Tyerman M J, Watson J E, Shane B, Schutz D E, Stokstad E L
Biochim Biophys Acta. 1977 Mar 29;497(1):234-40. doi: 10.1016/0304-4165(77)90156-8.
A simplified procedure for the determination of the glutamate chain lengths of endogenous tissue folate is described. Natural pteroylpoly-gamma-glutamates in tissue extracts, irrespective of their one-carbon moiety, were converted by a reductive cleavage procedure to a homologous series of p-aminobenzoylpoly-gamma-glutamates, differing only in glutamate chain length. Desalting and concentration of the extracts were achieved by absorbing the derivatives on active charcoal followed by their elution with ethanol:ammonia. Aminobenzoylpolyglutamates were separated by DEAD-cellulose chromatography and quantitated by a colorimetric procedure for primary aromatic amines. The major endogenous folates in rat liver and kidney were pteroylpentaglutamate derivatives with smaller amounts of pteroyltetra- and hexaglutamate.
本文描述了一种用于测定内源性组织叶酸谷氨酸链长度的简化方法。组织提取物中的天然蝶酰多聚-γ-谷氨酸，无论其一碳部分如何，通过还原裂解程序转化为一系列对氨基苯甲酰多聚-γ-谷氨酸同系物，它们仅在谷氨酸链长度上有所不同。提取物的脱盐和浓缩是通过将衍生物吸附在活性炭上，然后用乙醇：氨水洗脱来实现的。对氨基苯甲酰多聚谷氨酸通过DEAE-纤维素色谱法分离，并通过比色法对伯芳香胺进行定量。大鼠肝脏和肾脏中的主要内源性叶酸是蝶酰五谷氨酸衍生物，还有少量的蝶酰四谷氨酸和六谷氨酸。
