Simplified enzymatic high-performance anion exchange chromatographic determination of total fructans in food and pet food--limitations and measurement uncertainty

A simplified method to determine total fructans in food and pet food has been developed and validated. It follows the principle of AOAC method 997.08, i.e., high-performance anion exchange chromatographic (HPAEC) determination of total fructose released from fructans (F(f)) and total glucose released from fructans (G(f)) after enzymatic fructan hydrolysis. Unlike AOAC method 997.08, calculation of total fructans is based on the determination of F(f) alone. This is motivated by the inherent difficulty to accurately determine low amounts of G(f) since many food and pet food products contain other sources of total glucose (e.g., starch and sucrose). In this case, a correction factor g can be used (1.05 by default) to take into account the theoretical contribution of G(f). At levels >5% of total fructans and in commercial fructan ingredients, both F(f) and G(f) can and should be accurately determined; hence, no correction factor g is required. The method is suitable to quantify total fructans in various food and pet food products at concentrations greater than or equal to 0.2% providing that the product does not contain other significant sources of total fructose such as free fructose or sucrose. Recovery rates in commercial fructan ingredients and in selected food and pet food ranged from 97 to 102%. As part of a measurement uncertainty estimation study, individual contributions to the total uncertainty (u) of the total fructan content were identified and quantified by using the validation data available. As a result, a correlation between the sucrose content and the total uncertainty of the total fructan content was established allowing us to define a limit of quantitation as a function of the sucrose content. One can conclude that this method is limited to food products where the sucrose content does not exceed about three times the total fructan content. Despite this limitation, which is inherent to any total fructan method based on the same approach, this procedure represents an excellent compromise with regard to accuracy, applicability, and convenience.