@article{oai:kanazawa-u.repo.nii.ac.jp:00015063,
 author = {Koyama, Kazuo and Hirao, Takashi and Toriba, Akira and Hayakawa, Kazuichi},
 issue = {5},
 journal = {Biomedical Chromatography},
 month = {May},
 note = {The quality of starch-containing foods may be significantly impaired by contamination with very small amounts of α-amylase, which can enzymatically hydrolyze the starch and cause viscosity loss. Thus, for quality control, it is necessary to have an analytical method that can measure low amylase activity. We developed a sensitive analytical method for measuring the activity of α-amylase (from Bacillus subtilis) in starch-containing foods. The method consists of six steps: (1) crude extraction of α-amylase by centrifugation and filtration; (2) α-amylase purification by desalting and anion-exchange chromatography; (3) reaction of the purified amylase with boron-dipyrromethene (BODIPY)-labeled substrate, which releases a fluorescent fragment upon digestion of the substrate, thus avoiding interference from starch derivatives in the sample; (4) stopping the reaction with acetonitrile; (5) reversed-phase solid-phase extraction of the fluorescent substrate to remove contaminating dye and impurities; and (6) separation and measurement of BODIPY fluorescence by HPLC. The proposed method could quantify α-amylase activities as low as 10mU/mL, which is enough to reduce the viscosity of starch-containing foods. © 2012 John Wiley & Sons, Ltd.},
 pages = {583--588},
 title = {An analytical method for measuring α-amylase activity in starch-containing foods},
 volume = {27},
 year = {2013}
}