Enzymatic, spectrophotometric determination of glucose, fructose, sucrose, and inulin/oligofructose in foods

Abstract

A fast, simple, and accurate method, using only standard laboratory equipment, was developed for the quantification of glucose, fructose, sucrose, and inulin/oligofructose in different food matrixes. Samples were extracted using boiling water and hydrolyzed with sucrase and fructanase. Sugars were determined in the initial extract and in both hydrolysates using an enzymatic, spectrophotometric kit for glucose and fructose determination with hexokinase, glucose-6-phosphate dehydrogenase, and phosphoglucose isomerase. Calculations of sucrose and inulin/oligofructose were based only on fructose measurement. Glucose results of the hydrolysates were not used for inulin/oligofructose calculations because of possible interference. Released glucose by the hydrolysis of maltose or by possible partial hydrolysis of other compounds like maltodextrines, starch, lactose, or maltitol could interfere in the measurement of the sucrase and the fructanase hydrolysates. To validate the method, a wide range of different food matrixes and different amounts of inulin/oligofructose (1-54%) were analyzed. Mean recovery +/- relative standard deviation (RSD) for inulin or oligofructose was 96.0 +/- 5.3%. The RSDr for inulin/oligofructose measured on 35 food samples, analyzed in duplicate, was 5.9%. Accuracy and precision of the method were less for samples with large concentrations of sucrose, maltose, maltodextrines, or starch (ratio to inulin/oligofructose >4 to 1). Precision and accuracy were comparable with those of the ion exchange chromatographic method AOAC 997.08 and the enzymatic, spectrophotometric method AOAC 999.03. In contrast to 999.03, this method allows the accurate quantification of both GFn and Fn forms.