- Polymethoxyflavones (PMF's) are naturally occurring flavonoids present in Rutaceae family in the Citrus genus. Due to the potential use of PMF's as a chemopreventive agent and functional food, a rapid reproducible method for large scale isolation is critical. Isolation of PMFs from citrus is a challenging task due to complex sample matrices. Recent advances in chromatographic techniques led to several isolation and identification methods of PMF's from citrus. The present chapter discusses about the various techniques used for extraction and isolation of PMFs from different citrus species. The identification of these compounds using 1H NMR and 13C NMR is also described. © 2012 American Chemical Society.