Chlorophylls are essential for life because of their functional role in light harvesting for photosynthesis and photochemistry. It is the most abundant pigment on Earth, widely distributed in the tissues of all photosynthetic organisms, from algae to higher plants. Chlorophylls undergo chemical and/or enzymatic reactions during natural metabolism, food processing, and storage, being crucial to the accurate determination of the chlorophyll profile. This chapter describes a detailed experimental procedure and approach to extract chlorophylls from foods and their analysis with HPLC-UV/vis or HPLC hyphenated with MS to characterize the chlorophyll profile. The protocol includes information to facilitate the chlorophyll identification and the isolation of standards.