Authentication of L-Ascorbyl-2-sulfate in Salmonid Gastric Tissue: HPLC/Electrospray Ionization Mass Spectroscopic Verification

Abstract

New findings establish the intact transfer of the L-ascorbyl-2-sulfate ester across the stomach membrane in salmon. In addition, measurement of the ester has been confirmed in a commercial fish diet. The procedure used a non ion-pairing C18 reverse phase column for the separation of L-ascorbyl-2-sulfate (AS) from coho salmon gastric tissue. The AS fraction was eluted from the column with 0.1 M ammonium acetate pH 5.0 and rechromatographed to ascertain purity. When chromatographed under the same conditions, the collected AS fraction had the same retention time as the purified standard of L-ascorbyl-2-sulfate dipotassium salt. The purified AS fraction was then confirmed as to structure by Electrospray Ionization Mass Spectrometry The AS fraction gave the same Electrospray Ionization Mass Spectrometry results as the standard AS when subjected to mass spectrometry-mass spectrometry (MSMS) analysis.