A simple and rapid analytical UHPLC methodology with spectrophotometric (UV/Vis)\ndetection, coupled with different extraction procedures, has been perfected to investigate the presence\nof biologically active O-prenylated umbelliferone derivatives, such as auraptene and umbelliprenin,\nin pomegranate (Punica granatum L.) seed extracts. Absolute ethanol was the most efficient extraction\nsolvent in terms of yields, after a short ultrasound-assisted. The highest concentration values recorded\nunder these experimental conditions were 1.99 microg/g of dry extract and 6.53 microg/g for auraptene and\numbelliprenin, respectively. The parent metabolite umbelliferone was also detected (0.67 microg/g). The\nextraction and UHPLC analytical methodology set up in the present study proved to be an efficient,\npowerful, and versatile technique for the simultaneous qualitative analysis and quantification of\noxyprenylated coumarins in pomegranate seed extracts. The characterization of such secondary\nmetabolites in the mentioned phytopreparation represents, to the best of our knowledge, the first\nexample in the literature.
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