Metabolite profiling of polyphenols in peels of Citrus limetta Risso by combination of preparative high-speed countercurrent chromatography and LC–ESI–MS/MS

The polar constituents of peels from Citrus limetta variety Risso (Rutaceae) were investigated by a combination of two complementary chromatographic techniques consisting of preparative high-speed countercurrent chromatography (HSCCC), and off-line LC–ESI–MS/MS analysis to design a two-dimensional metabolite profile. Countercurrent chromatography (CCC) using solely immiscible solvent systems allowed the fractionation of principal components and an enrichment of minor concentrated metabolites from a crude polar solvent partition of C. limetta peels for subsequent structural identification by LC–ESI–MS/MS analysis. The combination of two very different chromatographic techniques resulted in lower detection limits for electrospray mass-spectrometry and revealed eighty-five compounds, including three abscisic acid derivatives, five limonoid glycosides, twenty-six dihydro-cinnamic and cinnamic acid glycosides, eleven flavanone glycosides, seven flavone glycosides, seventeen flavonol glycosides, including limocitrol and limocitrin derivatives. As a chemocharacteristic for C. limetta metabolites, many of the detected structures were linked to single and multiple 3-hydroxy-3-methyl-glutaryl (HMG) substitutions. C. limetta peels are a by-product of juice production, and not only the antioxidant fractions but also some of the fortified compounds could be used for food and pharmaceutical purposes.