Evaluation of DNA extraction procedures for traceability of various tomato products Manuela Turci, Maria Luisa Savo Sardaro, Giovanna Visioli, Elena Maestri, Marta Marmiroli, Nelson Marmiroli * Department of Environmental Science, Division of Genetics and Environmental Biotechnology, University of Parma, Viale G.P. Usberti, 11/A, 43100 Parma, Italy article info Article history: Received 28 November 2008 Received in revised form 20 April 2009 Accepted 26 April 2009 Keywords: DNA extraction methods Fluorimetric analysis PCR amplification SSR markers Fuzzy logic Solanum lycopersicum L. abstract Globalization of food trade requires the development of integrated approaches, such as traceability of ori- gin, quality and authenticity, to ensure food safety and consumers satisfaction. In this study, different genomic DNA extraction procedures were evaluated for their applicability to internal traceability of dif- ferent products in the tomato food chain. Quality, quantity and amplificability by SSR markers of extracted DNA tallied the methods performances; times and costs were considered too. The results were processed with ‘‘fuzzy-logic” approach. ‘‘Wizard” (Promega) scored the best performance in methods final ranking. This work demonstrated the value of genomic methodologies for internal traceability of tomato-derived goods. Ó 2009 Elsevier Ltd. All rights reserved. 1. Introduction The strategic development of a food chain approach to food quality and safety must be considered within a global context that is constantly evolving in terms of normative and requirements. Globalisation in food trade needs in particular the development of a more integrated and preventive approach. Expansion in international trade, highly integrated markets, more rapid adoption of new technologies, and an increased market concentration have important implications, both positive and neg- ative, for the development of a food chain approach to food safety (FAO, 2003). In particular, internal traceability has been indicated as a pro- duction action to improve reliability of labelling, to certify the ori- gin and the quality of products on the market, and to prevent fraudulent or deceptive labelling (EC No 178/2002). The European Union has considered the use of high-quality raw material in food production as a prerequisite to obtain a genuine and safe product of adequate nutritional value (White Paper on Food Safety. COM/ 99/719); consequently internal traceability is assuming a particu- lar relevance in the global process of traceability. The requirement of internal traceability procedures in food pro- duction has stirred also a certain level of technological implemen- tation (Di Bernardo, Del Gaudio, Galderisi, Cascino, & Cipollaro, 2007; Peano, Samson, Palmieri, Gulli, & Marmiroli, 2004). Method- ologies based on genetic and molecular biology are acquiring great interest for their applicability to track a given item at any stage along the food supply chain, from ‘‘farm to the fork” (Di Bernardo, Galderisi, Cipollaro, & Cascino, 2005). Among these, PCR analysis allows the identification of traces of genomic DNA that may resi- due in a food matrix from the principal component and/or from contaminants (Marmiroli, Peano, & Maestri, 2003). The DNA extraction method can affect the PCR based analysis by: (i) the presence of PCR inhibitors in the food matrices, (ii) the excessive fragmentation of the DNA molecules, and (iii) the short average length of DNA fragments. Quantity and quality of the extracted DNA are extremely sam- ple-dependent. In fact the food matrix production, and its chemi- co-physical composition can introduce many degrees of variability into the DNA extraction methods and in the efficacy of the DNA amplification (Di Bernardo et al., 2007; Peano et al., 2004). Moreover, a large number of plant species, including tomato, produce secondary metabolites such as phenolic compounds, tan- nins, flavonoids and alkaloids, whose presence in the extract can interfere with DNA analysis and inhibit its amplification (Di Ber- nardo et al., 2005). Processing at acid or alkaline pH may constitute 0956-7135/$ - see front matter Ó 2009 Elsevier Ltd. All rights reserved. doi:10.1016/j.foodcont.2009.04.012 * Corresponding author. Tel.: +39 0521 905606; fax: +39 0521 905402. E-mail addresses: manuela.turci@nemo.unipr.it (M. Turci), marialuisa.savosar- daro@unipr.it (M.L.S. Sardaro), giovanna.visioli@unipr.it (G. Visioli), elena.maestri@ unipr.it (E. Maestri), marta.marmiroli@unipr.it (M. Marmiroli), nelson.marmiroli@ unipr.it (N. Marmiroli). Food Control 21 (2010) 143–149 Contents lists available at ScienceDirect Food Control journal homepage: www.elsevier.com/locate/foodcont