Determination of Total, Saturated, Unsaturated, and Monounsaturated Fats in Cereal Products by Acid Hydrolysis and Capillary Gas Chromatography: Collaborative Study

Abstract

Fifteen laboratories participated in a collaborative study to determine total, saturated, unsaturated, and monounsaturated fats in cereal products by gas chromatographic (GC) analysis of fatty acid methyl esters (FAMEs). Cereal products, representing a wide range of cereal grains and processes, were hydrolyxed in 8N HCl and extracted with ethyl and petroleum ethers. FAMEs were produced by the reaction of the mixed ether extracts with sodium hydroxide in methanol (NaOH/MeOH) and then with boron trifluoride reagent (14% BF3 in MeOH). They were quantitatively determined by capillary GC. Total fat was calculated as the sum of individual fatty acids expressed as triglyceride equivalents in accordance with nutrition labeling guidelines. Saturated, unsaturated, and monounsaturated fats were calculated as sums of individual fatty acids. The total fat contents of samples ranged from 0.56 to 12.64%. A split design was used to determine performance parameters of results obtained by 15 laboratories on 24 samples. Of the 24 samples, 7 were blind duplicates and 5 were independent materials. Statistical analysis for total fat yielded a relative standard deviation for repeatability (RSDr) range of 1.32 to 13.30% and a relative standard deviation for reproducibility (RSDR) range of 4.42 to 22.82%. The goal of this study was to determine total fat, saturated fat, unsaturated, and monounsaturated fat in cereal-based products by complete extraction, methylation, and quantitation of total fatty acids. The acid hydrolysis-capillary GC method for determining total, saturated, unsaturated, and monosaturated fats in cereal products has been adopted by AOAC INTERNATIONAL.