Liquid chromatographic determination of L-ascorbate 2-polyphosphate in fish feeds by enzymatic release of L-ascorbate

An accurate method was devised to assay L-ascorbic 2-polyphosphate esters (AsPP) in fish feed by phosphatase digestion followed by determination of the released L-ascorbic acid (AsA). Compressed yeast and dithiothreitol are added to the phosphatase reaction mixture to give 95-100% recovery of AsA, which is quantitated by reverse-phase liquid chromatography (LC) with electrochemical detection. Chromatograms of all feed digests showed baseline resolution of AsA. In 3 feeds, to which 75-125 ppm AsA equivalents in the form of AsPP were added, the assay procedure gave 98-100% recovery of AsA.