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HPLC Determination of Lactulose in Heat Treated Milk

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Abstract

Lactulose (4-O-β-d-galactopyranosyl-d-fructose), formed from lactose (4-O-β-d-galactopyranosyl-d-glucose) by the Lobry de Bruyn–Alberda van Ekenstein rearrangement during severe heat treatments, is considered a useful indicator of heat-induced modifications in milk. Its chromatographic determination in milk is particularly troublesome due to the concomitant presence of a lactose amount two orders of magnitude larger than the lactulose amount and to a similar retention time of the two compounds. In this work, four HPLC methods have been compared with the aim to develop a more accurate analytical procedure to determine lactulose in milk, together with lactose. The developed method is based on a Carrez precipitation followed by a HPLC separation on two in-series amino-based columns, using CH3CN:H2O 75:25 (v:v) as the mobile phase at 1 ml/min flow rate and a refractive index as the detector. The linearity test for the quantitation of lactulose has been carried out over the range 0.060–1.006 mg/ml, the limit of detection, is 0.013 mg/ml (256 ng injected) and the limit of quantitation is 0.028 mg/ml (556 ng injected). The proposed method, simple, cheap and time-saving, allows an accurate lactulose–lactose separation, with conventional HPLC equipment.

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Acknowledgements

This research was financially supported by MiPAAF (Ministero delle Politiche Agricole, Alimentari e Forestali) Project: QUALIFU “Qualità Alimentare e Funzionale”. The authors are also grateful to anonymous referees and editor for helpful comments on an earlier draft. The authors thank Dr. L. Nachbin for editing of the manuscript and her valuable suggestions.

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  1. Istituto Nazionale di Ricerca per gli Alimenti e la Nutrizione, Food Science Area, Via Ardeatina, 546, 00178, Rome (I), Italy

    Pamela Manzi & Laura Pizzoferrato

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  1. Pamela Manzi
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  2. Laura Pizzoferrato
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Correspondence to Pamela Manzi.

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Manzi, P., Pizzoferrato, L. HPLC Determination of Lactulose in Heat Treated Milk. Food Bioprocess Technol 6, 851–857 (2013). https://doi.org/10.1007/s11947-011-0700-x

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