Apple (Malus pumila) procyanidins fractionated according to the degree of polymerization using normal-phase chromatography and characterized by HPLC-ESI/MS and MALDI-TOF/MS

Our previously reported method for the fractionation of apple procyanidins was modified successfully to achieve the separation of (epi)catechins and procyanidins (ranging from dimers to octamers) according to the degree of polymerization. Normal-phase chromatography was employed, using a hexane–methanol–ethyl acetate mixture as the mobile phase. Each fraction was characterized using high-performance liquid-chromatography electrospray-ionization mass spectrometry (HPLC-ESI/MS) and matrix-assisted laser-desorption ionization time-of-flight mass spectrometry (MALDI-TOF/MS). This method will be useful for the evaluation of the physiological functions of proanthocyanidins and for the elucidation of their individual structures.