Determinación de patulina en zumo de manzana por electroforesis capilar y cromatografía líquida de alta resolución. Incidencia y estimación de la ingesta por la población

Resumen

Patulin is a secondary metabolite produced by fungi from the genera Penicillium and Aspergillus. It is mainly found in apples. Its toxicicty in different animal species includes as main effect damage in the intestinal tract. In 1995, the JECFA/WHO recommended that human exposure to PAT should be reduced to less than 0.4 μg/kg body weight/day. The European commission approved a Directive (nº 1881/2006) which established PAT limits for different apple derivative commodities. In this work, the optimization and validation of analytical methods by capillary electrophoresis (CE) (3 different modes) for the determination of patulin in apple juice have been carried out. Moreover, a comparison has been made between the CE methods and the commonly used HPLC technique. During methods validations the following parameters were studied: selectivity, linearity, precision and accuracy, recovery, limits of detection and quantification, robustness and specific parameters for CE. A stability study of patulin in apple juice stored a -20ºC was carried out during a period of 6 months which guaranteed samples stability. This is the first time in which CE has been used for PAT determination in apple juice. The electrochromatography method did not fulfil the validation requirements because there were so many technical problems that the method reliability was compromised. Patulin was studied in samples of apple juice: acquired in Spain and intended for adults (n=100); purchased in Denmark and intended for adults (n=20); and obtained in Spain and intended for infants and babies (n=20). The analytical technique used was micellar electrokinetic chromatography. For these groups, the mean PAT levels obtained were 28.5, 35.9, 8.0 μg/L, respectively, being all below the maximum PAT level allowed by the legislation in force. With these mean values, patulin daily intake estimate was performed considering an apple juice consumption of 200 mL per day in adults (body weight: 70 kg), 200 mL per day in children (body weight: 25 kg) and 130 mL per day for babies (body weight: 10 kg). In each case, the estimations gave values below the provisional tolerable daily intake of PAT established by JECFA in 2005.