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Opuntia spp. (prickly pear) cultivation is playing a steadily growing role as a drought tolerant plant resource for fruit, and fodder production and also land conservation worldwide. From the view point of global climatical changes and the increasing needs for a consistent and reliable fruit production, cacti fruits are almost ideal crops for areas endangered by desertification and inconsistent precipitation levels. Various cacti plants have their origin from arid regions of North-America, and today they are distributed throughout most of all dry sub-tropical and tropical areas of the world. Betalain pigment profilings of betacyanins with their respective degradation products and betaxanthins occurring in yellow, orange and violet Opuntia fruits from Sicilia were done by using two chromatographic dimensions. Fractions recovered by preparative ion-pair HSCCC were investigated by off-line LC-ESI-MS/MS analysis.