The influence of different treatments, i.e., crushing, high temperature short time (90 degrees C/4 min) (HTST) and low temperature long time (60 degrees C/40 min) (LTLT) blanching, acidification (pH 4.3), and sequences of these treatments on the folate poly-gamma-glutamate profile and stability were investigated. In this study, broccoli was used as a case study. Regarding the folate poly-gamma-glutamate profile, endogenous folate poly-gamma-glutamates in broccoli florets were found predominantly as hepta- and hexa-gamma-glutamates. Crushing raw broccoli, acidification and LTLT blanching enhanced folate deconjugation resulting in monoglutamate, di- and tri-gamma-glutamates. Compared to other treatments, HTST blanching preformed prior to crushing resulted in the highest concentration of long chain poly-gamma-glutamates. Regarding folate poly-gamma-glutamates stability, acidification combined with LTLT blanching decreased folate stability whereas HTST blanching combined with different sequences of blanching and crushing did not affect folate poly-gamma-glutamates stability. It was concluded that crushing (prior to heating), acidification and blanching could be strategically applied to increase the folate monoglutamate content of broccoli. (C) 2009 Elsevier Ltd. All rights reserved.