A simple and rapid method using capillary zone electrophoresis (CZE) for the determination of milt protein (MP), which contains mainly protamine, and polylysine (PL) in food additive preparations and processed foods was developed. CZE separation was performed on poly(vinyl alcohol)-coated capillaries at a column temperature of 20῍ with 120 mmol/L phosphate bu#er (pH 2.5) as the running bu#er.The influence of various components in food additive preparations on CZE analysis of MP and PL was examined. Egg white lysozyme, glycine, sodium acetate, glycerol, fumaric acid, calcium carbonate, dextrin, emulsifiers and sodium polyphosphate and pyrophosphate had no e#ect. No peak of protamine was detected in preparations containing metaphosphate.The analysis method for processed foods was composed of extraction with 4῎ formic acid, precipitation of macromolecular compounds with ethanol, concentration in a water bath and determination by CZE. The average recoveries were 108.4῎ for protamine sulfate (PS) in red bean sticky rice, and 81.3῎ for PL in white rice, 118῎ in egg sandwiches, and 115῎ in shiraae. The limits of detection of PS in red bean sticky rice and PL in white rice were both 50 ppm.
Daily intake of isoflavones (daidzin, glycitin, genistin, daidzein, glycitein, and genistein) was determined quantitatively, based on the market basket method. Acid hydrolysis during extraction of foods was chosen to convert phytoestrogenes into the respective aglycons, facilitating HPLC analysis and allowing quantitation of total isoflavones as aglycones including both originally present glycosides and "free" aglycones. The isoflavones were extracted from samples with methanol and determined by reversed-phase HPLC analysis using a linear gradient of methanolῌ water as the eluent. From the results of hydrolysis, the daily intake of total isoflavon was 38.1 mg/adult Japanese.The values obtained by the market basket method and the National Nutrition Survey method were similar.
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