The EFSA Journal (2007) 481, 1-7

Transcription

1 The EFSA Journal (2007) 481, 1-7 Opinion of the Scientific Panel on Dietetic Products, Nutrition and Allergies on a request from the Commission related to a notification from IFF on mustard seed oil pursuant to Article 6, paragraph 11 of Directive 2000/13/EC (Request Nº EFSA-Q ) (adopted on 3 May 2007) SUMMARY Allergic reactions to mustard, including severe anaphylactic reactions, are well documented in clinical and laboratory studies. Mustard allergy is also encountered in children. Studies suggest that mustard allergy may account for 1-7% of all food allergies in some European regions. Mustard allergens are resistant to heat and to enzymatic degradation, and therefore are not markedly affected by food processing. This notification covers mustard-derived essential oil, i.e. mustard seed oil, used as flavour in foods. The applicant claims a safe use for allergic individuals on the basis of the typically low quantities of mustard oil in foods and of the low amount of mustard protein found in five tested mustard seed oil samples according to an ELISA test developed by the applicant (limit of detection 1.5µg/g). It should be noted that this test does not detect hydrophobic proteins preferentially present in mustard seed oil. Furthermore, the applicant has not performed any IgE in vitro test to detect allergenic proteins and fragments thereof, nor have new laboratory and/or clinical studies been provided in order to demonstrate the lack of allergenicity. The applicant does not take into consideration the scientific literature reporting adverse reactions to allyl isothiocyanate, which may cause immunologically-mediated adverse reactions and systemic symptoms upon a very low exposure. The incomplete scientific data submitted by the applicant do not allow the Panel to evaluate the likelihood that mustard seed oil will trigger an adverse allergic reaction in susceptible individuals. KEY WORDS Mustard oil, essential oil, allergenicity, ELISA test, allyl isothiocyanate. Page 1 of 7

2 BACKGROUND In November 2003, the European Parliament and the Council adopted Directive 2003/89/EC 1 amending Directive 2000/13/EC, as regards indication of the ingredients present in foodstuffs. Annex IIIa of the Directive specifies a list of food ingredients or substances that are known to trigger allergic reactions or intolerances in sensitive individuals for which no labelling exemptions are allowed. Whenever the listed ingredients/substances or their derivatives are used in the production of foodstuffs, they must be labelled. Article 1, paragraph 11, subparagraph 2 of the Directive establishes a procedure allowing for temporary labelling exemption of derivatives from ingredients listed in Annex IIIa for which it has been scientifically established that it is not possible for them to cause adverse reactions. In accordance with this provision, submissions of request for temporary labelling exemption were notified to the Commission before 25 August The Commission, after consultation with the European Food Safety Authority, adopted a list (Directive 2005/26/EC 2 ) of those ingredients which are temporarily excluded from Annex IIIa until 25 November 2007, pending the final results of the notified studies. Consequently, applicants who submitted a dossier in 2004 on the basis of subparagraph 2, resulting in the inclusion of a product in the list of Directive 2005/26/EC, and who are seeking exclusion of that product from Annex IIIa beyond 25 November 2007 will have to submit a request enclosing the final results of the notified scientific studies. Therefore in the context of the permanent labelling exemption procedure, the European Food Safety Authority is asked to provide scientific opinions on the submissions in accordance with the present terms of reference. TERMS OF REFERENCE In accordance with Article 29 (1) (a) of Regulation (EC) N 178/2002, the European Commission requests the European Food Safety Authority to evaluate the scientific data submitted by International Flavours & Fragrances (IFF) in the framework of the procedure laid down in Article 6, paragraph 11 of Directive 2000/13/EC. On the basis of that evaluation, EFSA is requested to issue an opinion on the information provided, and particularly to consider the likelihood of adverse reactions triggered in susceptible individuals by the consumption of the following ingredients/substances used under the conditions specified by the applicant: mustard seed oil. ASSESSMENT Allergic reactions to mustard, including severe anaphylactic reactions, are well documented in clinical and laboratory studies (Caballero et al., 2002; Figueroa et al., 2005). Mustard allergy is also encountered in children (Morisset et al., 2003). The larger studies with food-allergic 1 Directive 2003/89/EC of the European Parliament and of the Council amending Directive 2000/13/EC as regards indication of the ingredients present in foodstuffs. OJ L , p Commission Directive 2005/26/EC of 21 March 2005 establishing a list of food ingredients or substances provisionally excluded from Annex IIIa of Directive 2000/13/EC of the European Parliament and of the Council. OJ L 75, , p Page 2 of 7

3 patients suggest that mustard allergy may account for 1-7% of all food allergies with regional variations (NDA, 2004a). Mustard allergens are resistant to heat and to enzymatic degradation, and therefore are not markedly affected by food processing. The major mustard allergens identified are Sin a 1, belonging to the 2S albumin family, with a molecular weight (MW) of 14 kda (Menendez- Arias et al., 1988), and Bra j 1, also from the 2S albumin family, with a MW of approximately 16 kda (Gonzales de la Pena et al., 1991). A new major allergen in mustard seeds has been recently isolated and identified, an 11S globulin called Sin a 2 with a MW of 51 kda (Palomares et al., 2005), but not all mustard allergens and their occurrence in different species are known. In addition, allyl isothiocyanate is a major skin-sensitizing agent (non IgEmediated mechanism), which is also the major component of mustard oil (97-100%). Mustard protein allergic individuals may react to the protein content of the oil. Individuals sensitised to the skin-sensitising component allyl isothiocyanate may react to oil even in the absence of mustard proteins (Lerbaek, 2004). Taking account of the potential allergen content and well documented clinical allergic reactions in individuals sensitive to mustard (NDA, 2004a), it is appropriate for the Panel to assess the likelihood that mustard seed oil may cause an allergic reaction in mustard-allergic individuals. In 2004, the Panel issued an Opinion on a notification submitted by International Flavours & Fragrances (IFF) to the European Commission pursuant to Article 6, paragraph 11 of Directive 2000/13/EC as amended by Directive 2003/89/EC, for temporary exemption from labelling (NDA, 2004b). The study by Koppelman et al. (2007) does not contain new analytical data when comparing with the dossier submitted for temporary exemption. 1. Manufacturing process The starting material for the mustard oil produced by IFF is mustard seed from Brassica juncea grown in Canada. Mustard oil is identified by its CAS (Chemical Abstracts Service) Number: The production process involves several steps. Whole mustard seeds are ground. During the grinding, tap water is added to the seed and afterwards the slurry is incubated at 50 C for 30 minutes. During this step, the allyl isothiocyanate is released from its precursor by the liberated endogenous enzyme myrosinase. The released oil is removed by steam distillation under reduced pressure. The resulting distillate contains both water and volatile mustard oil. The mustard oil is separated from the water by centrifugation and dried using sodium sulfate. Subsequently, mustard oil is filtered and packed. 2. Characterisation of the product The applicant provides information regarding the quality control of the final oil. According to the analytical data obtained by gas-liquid chromatography, the main volatile component of mustard seed oil is allyl isothiocyanate (97-100%). Based on the available toxicological data, the applicant also indicates that this substance has been classified as toxic by inhalation, in Page 3 of 7

4 contact with the skin and if swallowed, and irritating to eyes, respiratory system and skin (Risk phrases R23/24/25/36/37/38/43) in Directive 92/32/EC. 2.1 Intended use According to the applicant, the mustard seed oil is intended to be used as such or as part of a flavouring preparation to deliver the taste of mustard to foods. Typical levels of mustard oil in foods (mayonnaise/dressing, soups and sauces, bouillons, savouries and snacks) as consumed vary between mg/kg. The applicant states that the use of mustard oil is self-limiting due to the powerful sensory impact of this flavouring. 3. Evidence of non-allergenicity The applicant claims that mustard oil is not likely to trigger adverse reactions on the basis of two arguments: 1) the typical low levels of mustard oil in foods, and 2) the in vitro demonstration that proteins are not present in amounts higher than 1.5 µg/g in five samples of mustard seed oil analysed with an ELISA test developed by the applicant. The applicant did not take into account the known toxicity of allyl isothiocyanate and its role in causing allergic contact dermatitis, or the possibility that proteins not detectable with the ELISA test could cause an IgE-mediated reaction (Lerbaek et al., 2004; Kohl and Frosch, 1990). 3.1 History of non-allergenicity of the product The anaphylactic reactions due to mustard seed allergens reported in the literature and in the previous Opinion of the Panel have not been considered (NDA, 2004a). There could be a significant under-reporting of reactions due to lack of awareness by allergic individuals of the presence of mustard oil in foodstuffs. In addition, the possibility of adverse reactions, including irritation to eyes, skin and respiratory system, due to allyl isothiocyanate (Brand and Jacquot, 2002) has not been addressed by the applicant. 3.2 Laboratory-based tests An ELISA test has been developed for the detection of mustard seed proteins. Mustard proteins were extracted from ground mustard seeds and the obtained extracts were used for immunisation of two rabbits. A classical immunisation protocol was used and antisera were raised to construct the ELISA. The specificity of the antisera for the various seed proteins present in the extract was evaluated using IgG-immunoblotting. An inhibition ELISA for mustard proteins was performed with a detection limit of 1.5 µg/g (Koppelman et al., 2007). Recoveries for the ELISA methodology are reported to vary, especially at low concentrations of analyte due to analytical imprecision. The antibodies employed for this ELISA test were raised against proteins extracted with an aqueous buffer (Tris), and therefore hydrophilic proteins. Out of all proteins in the mustard extracts, only three seem to be recognized by the antibodies, which have MWs different from those of the known allergens. Only hydrophilic proteins in mustard oil can be detected by this ELISA test. Any hydrophobic mustard protein, most likely present in mustard seed oil, will not have been detected using this ELISA test. Page 4 of 7

5 The applicant did not perform immunochemical analysis or IgE-binding studies with the mustard seed oil In vitro studies Using the above-described inhibition ELISA methodology, five samples of mustard seed oil from different batches were analysed looking at the potential presence of mustard proteins. The results showed that the protein content of all samples was below the detection limit of 1.5µg/g. On this basis, the applicant concludes that the use of this ingredient as flavouring in foods is unlikely to cause allergic reactions in mustard allergic individuals. The applicant does not take into consideration the possibility that proteins not detectable with this in vitro system could cause an IgE-mediated reaction Animal studies No animal studies were provided or referred to in the dossier. 3.2 Clinical studies No clinical studies have been provided by the applicant. A case report has been cited by the applicant (Morisset et al., 2003), which shows that one patient reacted to a dose of 0.8 mg of mustard protein in a placebo-controlled food challenge study. CONCLUSIONS The incomplete scientific data submitted by the applicant do not allow the Panel to evaluate the likelihood that mustard seed oil will trigger an adverse allergic reaction in susceptible individuals. DOCUMENTATION PROVIDED TO EFSA Dossier submitted by International Flavors & Fragrances (IFF) to the European Commission pursuant to Article 6, paragraph 11 of Directive 2000/13/EC as amended by Directive 2003/89/EC on 10 August REFERENCES Brand G and Jacquot L (2002). Sensitization and desensitization to allyl isothiocyanate (mustard oil) in the nasal cavity. Chem Senses 27: Caballero T, San-Martin MS, Padial MA, Contreras J, Cabanas R, Barranco P, Lopez-Serrano MC (2002). Clinical characteristics of patients with mustard hypersensitivity. Ann Allergy Asthma Immunol 89: Page 5 of 7

6 Council Directive 92/32/EEC of 30 April 1992 amending for the seventh time Directive 67/548/EEC on the approximation of the laws, regulations and administrative provisions relating to the classification, packaging and labelling of dangerous substances. Official Journal L 154, , p Figueroa J, Blanco C, Dumpierrez AG, Almeida L, Ortega N, Castillo R, Navarro L, Perez E, Gallego MD, Carrillo T (2005). Mustard allergy confirmed by double-blind placebocontrolled food challenges: clinical features and cross-reactivity with mugwort pollen and plant-derived foods. Allergy 60: Gonzalez de la Pena MA, Menendez Arias L, Monsalve RI, Rodriguez R (1991). Isolation and characterization of a major allergen from oriental mustard seeds, Bra J I. Int Arch Allergy Appl Immunol 96: Kohl PK and Frosch PJ (1990). Irritant contact dermatitis induced by mustard compress. Contact Dermatitis 23: Koppelman SJ, Vlooswijk R, Bottger G, van Duijn G, van der Schaft P, Dekker J, van Bergen H (2007). Development of an ELISA method to detect mustard protein in mustard seed oil. J Food Prot 70: Lerbaek A, Rastogi SC, Mennè T (2004). Allergic contact dermatitis from allyl isothiocyanate in a Danish cohort of 259 selected patients. Contact Dermatitis 51: Menendez Arias L, Moneo I, Dominguez J, Rodriguez R (1988). Primary structure of the major allergen of the yellow mustard (Sinapis alba L.) seed, Sin A 1. Eur J Biochem 177: Morisset M, Moneret-Vautrin DA, Maadi F, Fremont S, Guenard L, Croizier A, Kanny G (2003). Prospective study of mustard allergy: first study with double-blind placebo-controlled food challenge trials (24 cases). Allergy 58: NDA (Scientific Panel on Dietetic Products, Nutrition and Allergies) (2004a). Opinion of the Scientific Panel on Dietetic Products, Nutrition and Allergies on a request from the Commission relating to the evaluation of allergenic foods for labelling purposes. The EFSA Journal 32, NDA (Scientific Panel on Dietetic Products, Nutrition and Allergies) (2004b). Opinion of the NDA Panel related to a notification from IFF on mustard seed oil pursuant to Article 6 paragraph 11 of Directive 2000/13/EC. The EFSA Journal 157, Palomares O, Cuesta-Herranz J, Rodríguez R, Villalba MA (2005). Recombinant precursor of the mustard allergen Sin a 1 retains the biochemical and immunological features of the heterodimeric native protein. Int Arch Allergy Immunol 137: Page 6 of 7

7 PANEL MEMBERS Jean-Louis Bresson, Albert Flynn, Marina Heinonen, Karin Hulshof, Hannu Korhonen, Pagona Lagiou, Martinus Løvik, Rosangela Marchelli, Ambroise Martin, Bevan Moseley, Andreu Palou, Hildegard Przyrembel, Seppo Salminen, John (Sean) J Strain, Stephan Strobel, Inge Tetens, Henk van den Berg, Hendrik van Loveren and Hans Verhagen. ACKNOWLEDGEMENT The Scientific Panel on Dietetic Products, Nutrition and Allergies wishes to thank Taraneh Dean, Martin Stern, Jean-Michel Wal, and Adrianus van Hengel for their contributions to the draft opinion. Page 7 of 7