Fructan content determination by HPLC method with refractometric detection
2014
Bohacenko, I., Vyzkumny Ustav Potravinarsky, Prague (Czech Republic) | Pinkrova, J., Vyzkumny Ustav Potravinarsky, Prague (Czech Republic)
The method for fructan content determination using HPLC with refractometric detection and NH2 column has been implemented and internally validated. As well as other method, this method is based on the balance of glucose and fructose before and after hydrolysis of fructans by inulinase. In the first phase of testing under optimized chromatographic conditions, very good properties of NH2 column for separation of glucose, fuctose, saccharose, 1-kestose, nystose and 1-F-fructofuranosylnystose mixture have been demonstrated. During the internal validation of fructan determination in standard material, standard deviation sr = 0.2; expanded uncertainty of measurement U = +/- 0.4 % and mean recovery 97.9 % were determined. Standard deviations of fructan determinations in commercial materials Orafti P95, Orafti GR and Actilight 950P were very low, i.e. 0.04 and 0.05 respectively, which corresponds to the extended uncertainty of measurements U = +/- 0.1 %. Mean recovery of fructan determination, indicatively assessed by comparison with the fructan contents declared by the producer, ranged from 96.999.3 %. The method also allows a good separation and identification of individual fructan oligomers with DP 26 (both types GFn and Fn). These properties can be used to distinguish between different kinds of fructans. It can be concluded that the proposed method gives good prerequisites for the determination of fructans. After verification on real food samples, this method may be a good alternative to the official AOAC 997.08 method, especially in laboratories which do not have HPAEC-PAD chromatographic technology at their disposal.
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