Development and Optimisation of Solid-Phase Extraction of Extractable and Bound Phenolic Acids in Spelt (Triticum spelta L.) Seeds

A solid-phase extraction (SPE) technique was developed and optimised for isolation and concentration of extractable and bound phenolic acids from germinated spelt seeds, for analysis by liquid chromatography–mass spectrometry. Samples initially underwent solvent extraction under different conditions to maximise the yield of phenolic antioxidants. Optimal extraction conditions for extractable phenolics were absolute methanol as solvent, sample-to-methanol ratio 1:9, and reconstitution in non-acidified water. The bound phenolics were extracted from sample pellets using hydrolysis with 2 M NaOH, acidification of the hydrolysate with formic acid, and simultaneous isolation and purification using Strata X polymeric RP tubes. Compared to liquid-liquid extraction, this direct SPE protocol has significant advantages in terms of higher extraction efficiencies of total and individual phenolics and their antioxidant activities. These data suggest that direct SPE represents a rapid and reliable method for quantitative analysis of both the extractable and the commonly overlooked bound phenolics in Triticum spelta seeds.