The importance of medicinal products originated from the plant materials has greatly increased in recent years. Among the different bioactive natural products, flavonoids are crucial compounds and are represented by over 8000 varieties. These flavonoid substances are found largely in the foods of plant sources, and they are subdivided into 13 categories including anthocyanidins, catechins, flavonols and flavones. Considering the promising protective effects of flavonoids against cardiovascular diseases and certain types of cancers, increased research has been conducted for the extraction, purification and quantification of distinct flavonoid compounds. Therefore, this chapter intends to furnish a comprehensive view on the advanced extraction, cleanup and quantification techniques that are reported for the improved recovery of different flavonoids from medicinal plants, vegetables and fruits. Several non-conventional techniques such as microwave-assisted extraction, ultrasound-assisted extraction, pressurized-liquid extraction and supercritical fluid extraction processes reported for the improved extraction of flavonoids were critically examined. Further, different analytical methods recently reported for the separation and quantification of flavonoids were discussed extensively. At the end, conclusions and possible recommendations that enhance the extraction and quantification performance of flavonoids were highlighted. © 2020 Elsevier B.V.