Application of LC–MS–MS to identify niacin in aleurone layers of yellow corn,barley and wheat kernels

Previous studies identified type II inclusions in-situ of aleurone layers as niacin reserves and quantified niacin using colorimetric methods. Such identification and quantification was based on the König reaction which lacks specificity because cyanogen bromide reacts with all pyridine derivatives. The aim of this investigation was to define the structure of niacin in aleurone layers and aleurone cell contents of yellow corn, wheat and barley using LC–MS/MS. Aleurone layers were manually separated and cell contents released by ultrasonic processing and the residue and pellets were examined microscopically. Niacin was extracted from the samples by autoclaving in alkali. The extracts were analysed using HPLC and the structural identity of niacin was confirmed using quadrupole time of flight mass spectrometry in positive mode. Niacin concentrations were highest in wheat aleurone and lowest in corn aleurone. The MS/MS spectra of pure niacin and the purported niacin peak in sample extracts showed similar fragmentation patterns. The major ion product occurred at m/z 80 representing loss of CO₂ and parent ion at m/z 124 (M + H)⁺. These findings define some of the structural characteristics of niacin in aleurone layer of cereal grains, and demonstrate the possibility of using an ultrasonic processor to release cell contents.