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A highly sensitive method for the separation and quantitation of tetrahydrofolylpolyglutamates (H 4 PteGlu n  ) and their 5-methyl derivatives (CH 3 H 4 PteGlu n ) of different chain lengths by a reversed phase high-performance liquid chromatography with electrochemical detection is described. These compounds have been separated using a cosmosil 5-Ph column eluted with 50 mM potassium dihydrogen phosphate, pH 2.2, containing 0.05 M disodium EDTA and 1 % methanol at a tlow-rate of 1.5 ml/min, and an electrochemical detector (at a potential of + 350 mV versus the Ag/AgCI electrode). Good separation has been obtained on both H4PteGlun and CH 3 H 4 PteGIu n  derivatives containing up to 6 glutamyl residues. The detection limits at a signal-to-noise ratio of 3 for H 4 PteGlu n  and CH 3 H 4 PteGIu n  series (n = 1 - 6) were 1 -10 and 10 -15 pmol, respectivel y.

pteridines

Separation and Determination of Tetrahydrofolylpolyglutamates and their 5-Methyl Derivatives by Reversed-phase High-performance Liquid Chromatography with Electrochemical Detection