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Olive Oil Adulterated with Hazelnut Oils

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Olive Oil Adulterated with Hazelnut Oils Olive oil adulterated with hazelnut oils: simulation to identify possible risks to allergic consumers Marco Arlorio, Jean Daniel Coisson, Matteo Bordiga, Fabiano Travaglia, Cristiano Garino, Laurian Zuidmeer, Ronald van Ree, Maria Gabriella Giuffrida, Amedeo Conti, Aldo Martelli To cite this version: Marco Arlorio, Jean Daniel Coisson, Matteo Bordiga, Fabiano Travaglia, Cristiano Garino, et al.. Olive oil adulterated with hazelnut oils: simulation to identify possible risks to allergic consumers. Food Additives and Contaminants, 2009, 27 (01), pp.11-18. 10.1080/02652030903225799. hal-00572615 HAL Id: hal-00572615 https://hal.archives-ouvertes.fr/hal-00572615 Submitted on 2 Mar 2011 HAL is a multi-disciplinary open access L’archive ouverte pluridisciplinaire HAL, est archive for the deposit and dissemination of sci- destinée au dépôt et à la diffusion de documents entific research documents, whether they are pub- scientifiques de niveau recherche, publiés ou non, lished or not. The documents may come from émanant des établissements d’enseignement et de teaching and research institutions in France or recherche français ou étrangers, des laboratoires abroad, or from public or private research centers. publics ou privés. Food Additives and Contaminants For Peer Review Only Olive oil adulterated with hazelnut oils: simulation to identify possible risks to allergic consumers Journal: Food Additives and Contaminants Manuscript ID: TFAC-2009-194.R1 Manuscript Type: Original Research Paper Date Submitted by the 29-Jul-2009 Author: Complete List of Authors: Arlorio, Marco Coisson, Jean Daniel Bordiga, Matteo; School of Pharmacy, Università del Piemonte Orientale "A. Avogadro", Discaff Travaglia, Fabiano Garino, Cristiano Zuidmeer, Laurian van Ree, Ronald Giuffrida, Maria Conti, Amedeo Martelli, Aldo Methods/Techniques: Chromatography - electrophoresis, Immunoassays Additives/Contaminants: Allergens Food Types: Olive oil http://mc.manuscriptcentral.com/tfac Email: [email protected] Page 1 of 23 Food Additives and Contaminants 1 2 3 4 Olive oil adulterated with hazelnut oils: simulation to identify possible 5 6 risks to allergic consumers 7 8 9 10 11 M. ARLORIO 1, J.D. COISSON 1, M. BORDIGA 1, F. TRAVAGLIA 1, C. GARINO 1, L. ZUIDMEER 2, R. 12 13 VAN REE 2, M.G. GIUFFRIDA 3, A. CONTI 3 & A. MARTELLI 1 14 15 16 For Peer Review Only 17 1DiSCAFF and Drug and Food Biotechnology Center, Via Bovio 6, 28100 Novara (Italy) 18 19 2Department of Experimental Immunology and Department of Otorhinolaryngology, Academic Medical 20 21 Center, Meibergdreef 9, NL-1105 AZ Amsterdam (The Netherlands) 22 23 3ISPA-CNR, Via Ribes 5 - 10010 Colleretto Giacosa (Italy) 24 25 26 27 Corresponding Author: 28 29 Marco Arlorio 30 31 Discaff, Via Bovio 6, 28100 Novara (Italy) 32 33 Tel. +390321-375772 34 35 Fax +390321-375621 36 37 [email protected] 38 39 40 41 42 43 44 45 46 47 48 49 50 51 52 53 54 55 56 57 58 59 60 http://mc.manuscriptcentral.com/tfac Email: [email protected] Food Additives and Contaminants Page 2 of 23 1 2 3 4 5 ABSTRACT 6 7 8 9 10 To be labelled as “extra-virgin”, olive oil should be cold-pressed and may not contain refined oil or 11 12 oil from other oleaginous seeds or nuts (Reg. EC 1531/2001). Adulteration of extra virgin olive oil 13 14 (EVOO) with hazelnut oil (HAO) is a serious concern both for oil suppliers and consumers. The 15 16 high degree of similarityFor between Peer the two fatsReview complicates the detectionOnly of low percentage of HAO 17 18 in EVOO. Many analytical approaches have been developed during last years to trace HAO in 19 20 21 EVOO, principally based on chromatographic analyses, differential scanning calorimetry and 22 23 nuclear magnetic resonance. Adulteration of EVOO with HAO may introduce hazelnut-derived 24 25 allergens. The aim of this work was to analyse the protein and allergen content of EVOO 26 27 intentionally spiked with raw cold-pressed HAO or solvent-extracted HAO. SDS-PAGE analysis 28 29 confirmed the presence of hazelnut proteins in solvent-extracted HAO with molecular masses 30 31 ranging from 10-60 kDa. In contrast, cold-pressed HAO did not show traces of protein. In spiked 32 33 34 EVOO, solvent-extracted HAO was still detected at 1% contamination level. Several bands on 35 36 SDS-PAGE migrated at apparent molecular masses coinciding with known allergens like Cor a 1 37 38 (~17 kDa), Cor a 2 (~14 kDa), Cor a 8 (~12 kDa), oleosin (~17 kDa) and Cor a 9 (~60kDa). MALDI- 39 40 TOF MS analysis confirmed the presence of two oleosin isoforms and of Cor a 9. Immunoblotting 41 42 demonstrated that an allergic patient with known reactivity to Cor a 1 and Cor a 2 indeed 43 44 recognized a 17 kDa band in solvent-extracted HAO. In conclusion, we have shown that 45 46 47 adulteration of extra virgin olive oil with solvent extracted hazelnut oil can be traced by simple 48 49 SDS-PAGE analysis, and that such practice introduces a potential risk for hazelnut allergic 50 51 patients. 52 53 54 55 Keywords:- Hidden allergens, olive oil, hazelnut oil, oleosins. 56 57 58 59 60 http://mc.manuscriptcentral.com/tfac Email: [email protected] Page 3 of 23 Food Additives and Contaminants 1 2 3 4 5 Introduction 6 7 8 Due to its nutritional and commercial value, the consumption of olive oil has increased 9 10 considerably over recent years, representing 15.2% of the total oil utilization in the European Union 11 12 (Firestone 1985). Production of olive oil represents about 3% of the world oils and fat markets, with 13 14 78% of global production taking place in Mediterranean countries, where oil industry is important in 15 16 sustaining viabilityFor of agricultural Peer economy. Review The cultivation, harvesting Only and extraction of olive oil is 17 18 a hard and time-consuming task that adds to its relatively high commercial price. Olive oil is either 19 20 21 classified as “virgin olive oil” (i.e. cold-pressed olive oil produced by the use of physical means 22 23 without chemical treatment), as “olive oil” (i.e. a mixture of virgin and refined oil) or “refined oil” (i.e. 24 25 olive oil chemically treated to neutralize strong taste and in particular acidity) (Reg. EC 1531/2001). 26 27 Oils labelled as “extra-virgin olive oil” (EVOO) must not contain any refined olive oil; oils labelled as 28 29 “olive oil” must not contain oil from other oleaginous nuts or seeds. To increase profits, 30 31 unscrupulous dealers may be tempted to add lower-priced vegetable or hazelnut oils to fresh extra 32 33 34 virgin olive oils. This fraudulent practice, in addition to being unfair to the consumer in terms of the 35 36 cost (Chiavaro 2008), may also cause severe health and safety problems such as the occurrence 37 38 of Spanish toxic oil syndrome in 1981 (Gelpi 2002, Blanch 1998). 39 40 41 42 Hazelnut oil (HAO), obtained from Corylus avellana L. seeds, is often used in extra-virgin olive oil 43 44 (EVOO) adulteration, because of the similar chemical composition with respect to both major and 45 46 47 minor compounds (Vichi 2001). The high degree of similarity complicates the detection of HAO in 48 49 EVOO, especially at concentrations below 20% (Parcerisa 1998; Ozen 2002) . The seriousness of 50 51 the problem has been aggravated during the last twenty years. In 1993/94 1,829,000 tonnes of 52 53 olive oil were consumed world wide. In 1995 it has been reported that quantities of Turkish 54 55 hazelnut oil was illegally introduced into the European community, without proper declaration to 56 57 customs and excise, in order to adulterate olive oil (Aparicio 2004). The loss to European 58 59 60 consumers was estimated at 40 million euros due to hazelnut oil being three times as cheap as olive oil. In 1997 the European Commission, Unit for the Coordination of Fraud Prevention http://mc.manuscriptcentral.com/tfac Email: [email protected] Food Additives and Contaminants Page 4 of 23 1 2 3 (UCLAF), estimated that more than 20,680 tonnes of hazelnut oil was used for producing 103,400 4 5 tonnes of adulterated olive oil (UCLAF 1997; European Union Research Committee 2001). 6 7 8 Many analytical approaches have been developed during last years in order to detect HAO in 9 10 EVOO, principally based either on chromatographic analyses of free and esterified sterols, or on 11 12 the detection of other minor secondary compounds, like filbertone (( E)-5 methylhept-2-en-4-one) 13 14 (Parcerisa 2000; Flores 2006a; Flores 2006b). Other advanced approaches such as differential 15 16 scanning calorimetry,For nuclear Peer magnetic Review resonance and isotop Onlyic ratio coupled with mass 17 18 spectrometry were also described (Chiavaro 2008; Sacchi 1997; Remaud 1997). Until today, no 19 20 21 validated PCR-based methods for identifying fraudulently spiked HAO in EVOO have been 22 23 reported. 24 25 26 27 HAO, like other edible oils, is largely considered a “protein-free” food but solvent-assisted 28 29 extraction for “industrial” oil can potentially (and probably) influence the presence of residual 30 31 hazelnut proteins, as has been shown for other nuts and legume oil preparations (Crevel 2000). 32 33 34 Some evidence highlighted that peanut oil (as well as the soybean oil) can not always be 35 36 considered as a safe product for peanut-allergic patients (Hourihane 1997; Crevel 2000; Ring & 37 38 Möhrenschlager 2007). Apart from a recent paper focused on the use of a biosensor (Bremer 39 40 2009), no specific data are available for protein content of either raw or refined HAO.
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