Cereal Chem. 77(5):685-691The size-exclusion (SE) HPLC profile of total protein extract obtained by sonication of flour samples at ambient temperature showed marked instability on reinjection. Instability was related to the presence of flour proteases that were inactivated by thermal treatment of flour samples at 60°C. Extraction of flour protein by sonication was a function of ultrasonic energy (sonication time × power product) delivered to flour sample. As protein solubility increased, the proportions of the earliest eluted SE-HPLC fractions (F1 and F2) increased. Oversonication of proteins evidenced by a decrease in F1 amount at the benefit of F2 occurred below the sonication energy level needed for total protein extraction. Ultrasonic energy level was adjusted to allow total protein extraction while limiting oversonication. The sonication procedure was applied on 27 flour samples of contrasting dough strength to extract total proteins. Absolute amount of protein extractable by sonication, determined from SE-HPLC area, was strongly correlated with flour protein content. Very significant and equivalent relationships were found between alveographic W index and absolute amount of either unextractable protein extract or F1 of SE-HPLC profile from total protein extract.