The EFSA Journal (2005) 185, 1-7

Transcription

1 The EFSA Journal (2005) 185, 1-7 Opinion of the Scientific Panel on Dietetic Products, Nutrition and Allergies on a request from the Commission related to a notification from DWV on milk products, egg products and fish products used in the manufacture of wine pursuant to Article 6 paragraph 11 of Directive 2000/13/EC (Request N EFSA-Q ) (adopted on 22 February 2005) SUMMARY This notification by Deutscher Weinbauverband E.V. (DWV) deals with several different wines, that are not well characterised either in terms of geographic origin, varieties of grapes, or in terms of the conditions of treatments applied during the wine making process. This notification also deals with several food allergens, i.e. fish products such as gelatine and isinglass; egg products such as albumin and lysozyme; and milk products such as casein, that are used as processing aids for making wine and may partly remain in the finished products. All the substances specified are known major allergens with a possible exception for the fishderived product isinglass. The application provides no consistent information on the residual level of proteins from the fining agents in the finished product. The evidence given of the non-allergenicity of the fined wines is weak. The historical review is not documented, the strategy of search is not detailed and the absence of known cases of allergic reaction to fined wines may also be due to under-reporting and lack of awareness of the fining agents used in the wine making process. The currently available experimental data are preliminary and not sufficient to exclude residual allergenicity. The scientific data submitted by the applicant do not allow the Panel to predict the likelihood of adverse reactions in milk, egg and fish allergic individuals. Appropriate analytical and clinical studies applying good clinical and good laboratory practice should be carried out. KEY WORDS Wine, processing aids, isinglass, fish gelatine, egg albumin, lysozyme, casein. 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. 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 Page 1 of 7

2 Annex IIIa of the Directive specifies a list of ingredients that are known to trigger allergic reactions or intolerances for which no labelling exemptions are allowed. Whenever the listed ingredients or their derivatives are used in the production of foodstuffs, they must be labelled. Article 1, paragraph 11 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 shall, not later than 25 November 2004, and after consultation with the European Food Safety Authority, adopt a list of those ingredients which shall be temporarily excluded from Annex IIIa, pending the final results of the notified studies, or at the latest until 25 November Therefore, 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 Deutscher Weinbauverband E.V. (DWV) in the framework of the procedure laid down for temporary labelling exemptions 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, pending the final results of the studies undertaken, 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: isinglass, fish gelatine; albumin, lysozyme (produced from eggs); casein (produced from milk). ASSESSMENT Alcoholic beverages such as wines are included in the new regulation on food labelling. Processing aids used for making wine which may partly remain in the finished products (i.e. red and white wines) need to be labelled when they derive from food allergens listed in Annex IIIa of the Directive. This notification deals with several food allergens, i.e. fish products such as gelatine and isinglass; egg products such as albumin and lysozyme and milk products such as casein that are used as processing aids for making wine. All the substances concerned are known major allergens (NDA, 2004) with a possible exception for the fish-derived product isinglass. 1. Manufacturing process Detailed information is provided on the origin, manufacturing and characteristics of the allergenic ingredients but the dossier does not provide a detailed description of the manufacturing process for making wines and does not make any reference to good manufacturing practices. Page 2 of 7

3 The applicant refers to different wines (red and white wines) that are produced in different regions. The principle of fining is to mechanically remove insoluble and colloidal substances from wine. Isinglass (a fish collagen preparation), fish gelatine, casein and egg white/albumin act as coagulating agents and their addition to wines or must results in the precipitation of those substances. The applicant states that normally, after a clarification process, the wine is filtered afterwards. Lysozyme is used for its anti-bacterial effect in order to prevent the growth of anaerobic bacteria and to better control the fermentation process. No detailed analytical data are given on whether or how the fining agents are completely removed after the clarification process, or to what level. 2. Product characterisation A detailed description of the products used as processing aids is given. Complete information is provided on the characteristics, origin, identification, manufacturing process, specifications of the fining agents used, i.e. i) fish products: isinglass, fish gelatine; ii) egg products: albumin, lysozyme; and iii) milk products: casein. The allergenic ingredients used as processing aids are well characterised and their quality seems to be controlled. The applicant provides only very general indications with regard to the intended use. No exposure estimations are given. 2.1 Isinglass Isinglass is a form of fish-derived collagen. The collagen of isinglass forms a positively charged colloidal solution at the ph level of wine. The clarification is achieved by neutralizing the negatively charged particles in suspension in wine which then aggregate and sediment. Generally isinglass is used at a concentration of 1 g/hl of wine. After 8-14 days, the wine can be filtered. The applicant states that no scientific publications on isinglass residual levels in wine exist and that the conventional use of isinglass always attempts to remove the fining agent completely. 2.2 Fish gelatine Gelatine is traditionally used to clarify wines and reduce tannin levels. Gelatine is made by denaturation of collagen, which is derived from fish. Fish gelatine has a different structure but the amino acid composition is similar to that of isinglass. Thirty to 100 g of fish gelatine is needed for 1 hl of wine (i.e mg/l). No data about the final concentration in wine are provided by the applicant. 2.3 Egg albumin Page 3 of 7

4 The main egg white protein component is ovalbumin which is a glycophospho-protein, responsible for the fining effect. Albumin is produced from fresh hen s egg white by pasteurisation and spray drying. Egg white is used to remove the tannins from red wine. Albumin is also mainly used to improve the sensorial qualities of red wine, e.g. colour, taste, acidity. Like some other animal proteins (e.g. fish collagen and gelatine), albumin neutralizes and forms aggregates with negatively charged particles in suspension in wines that thus precipitate. Egg white is used at a concentration of g/hl for red wine. No data about residual concentration of egg protein in wine are provided by the applicant. 2.4 Lysozyme Lysozyme (mucopeptide N-acetylmuramyl-hydrolase) is a 129 aminoacid polypeptide which is found in various plant and animal tissues, particularly in milk and egg white. It is an enzyme with an anti-microbial activity against in particular Gram positive bacteria. It allows to better control the fermentation process and to avoid the formation of volatile acids. It prevents the development of lactobacilli and thus increases the effect of sulphites that can be used in lower quantities when lysozyme is added. The recommended dose varies from to 50 g/hl to prevent or to stop bacterial growth and malolactic fermentation in must and wine. 2.5 Casein Casein is a mixture of phosphoproteins. Whole casein is the major protein fraction of milk and constitutes the coagulum or curd (i.e. the solid fraction of proteins obtained after coagulation of milk) which is formed by action of chymosin (rennin) or by acidification of the milk to ph 4.6. Caseins are characterised by clusters of hydrophobic and hydrophilic amino acids; this amphiphilic character is responsible for the structure and properties of caseins. The different caseins cross-link to form ordered aggregates, micelles, in suspension in lactoserum. Casein coagulates in the acidic environment of must and wine. In addition it binds and eliminates phenolic compounds, particularly tannins that may affect wine colour and taste. The fining effect is based on coagulation due to the same mechanisms as described for the other animal proteins. The usual dose ranges between 10 to 20 g/hl potassium caseinate but may be increased up to 60 g/hl. No data about the residual concentration in wine are provided by the applicant. 3. Evidence of non-allergenicity 3.1 History of non-allergenicity of the product The applicant provides a general review on food allergens and particularly on fish, egg and milk allergens. Regarding the specific products derived from fish, eggs and milk used as processing aids in this application and regarding the specific finished products that are concerned, reference to historical reviews is provided. However no information is given by the applicant on the basis of the statement that: Page 4 of 7

5 - Isinglass/gelatine is used for hundreds of years to clarify wine and beer, so far, not a single case of fish allergy related to this substance is known, and that in the medical literature, there is no indication of the allergenic potential of wine that has been fined with isinglass or fish proteins. - In the medical literature, there is no indication of the allergenic potential of wine that has been fined with egg products. - In the medical literature, there is no indication of an allergenic potential of wine that has been fined with milk products. The level of evidence for these assumptions is low since the strategy and the sources used for the search are not detailed and also because the risk is high that allergic reactions to fish, egg or milk are not diagnosed when adverse reactions due to consumption of wines are observed and that such allergic reactions are thus under reported. 3.2 Laboratory-based tests In vitro studies Analytical studies The applicant provides a general review on analytical methods that are available and appropriate for the detection of allergens and particularly on those that have enough sensitivity as to detect residual levels in wines. A major issue for the intended application is the quantitative analysis of residual amounts of the fining agents that may remain in wines. The applicant refers to a proposal by the Australian Grape and Wine Research and Development Corporation (GWRDC) whose research project combines analytical and clinical studies. However, the applicant does not seem to be associated with this study. On behalf of the German Wine Growing Association a pilot study was performed. Two white wines (Riesling, Mosel-Saar-Ruwer; Pinot blanc, Baden) and one red wine (Dornfelder, Pfalz) were treated with the processing aids at maximum doses of 50 g/hl of milk potassium caseinate, 3 g of isinglass and 12 g of ovalbumin. The maximum dose was approximately 3 times higher than that normally used in the wine processing. For each wine the protein composition was analysed by electrophoresis and silver staining. In the fined wines the applicant states that the typical bands for isinglass could not be detected. Similarly, neither the specific bands of ovalbumin and egg white products nor a band that would be typical for potassium caseinate were detected in any of the various fined wines IgE binding studies After the electrophoretical separation of the proteins and in parallel with their silver staining in the studies above, the allergens present in the fined wines were revealed by western blotting using a poorly characterised pool of sera of allergic patients sensitised to fish, eggs or milk. Negative controls were done with sera from non-allergic individuals. Positive controls were done with the fining agents and with fish, egg or milk extracts using the sera from Page 5 of 7

6 allergic patients. As stated by the applicant, none of the wines showed the typical bands of the fining agents derived from fish, eggs or milk during immunodetection procedures. As stated by the applicant this is a preliminary pilot study which is not sufficient to clearly establish the absence of allergenicity in fined wines. In particular the protein bands corresponding to potential allergens have not been precisely characterised but only identified by reference to protein databases. No quantitative evaluation has been made of the sensitivity of the assays and in particular no consistent clinical and biological information is provided on the patients recruited for the study. Sera are only partially characterised and, in addition, they have been pooled and not individually used which precludes detecting any reaction in a particular patient who would be particularly highly sensitised to a protein of the fining agents Animal studies The applicant mentions a study where animal models are used. However, no details are given. 3.3 Clinical studies No clinical studies were provided by the applicant. 3.4 Proposed studies The outlines of a study which is planned to provide better information on the allergenic potential of the final wines are presented by the applicant. This study includes the development of analytical methods for detection and characterisation of the proteins of the fining agents and for the quantification of their residues in wines. It also includes the development of in vitro testing and IgE binding studies and the development of clinical studies to be carried out with patients with relevant allergies. The applicant describes future studies regarding identification and quantification of the proteins of wines and of the fining agents using mass spectrometry in association with liquid chromatography; development of improved sensitive and specific ELISA tests for the different proteins of the fining agents and particularly for isinglass; development of electrophoretic separation of proteins of wines and of the fining agents and immunological analysis by western blotting using sera of patients allergic to fish, egg or milk. The applicant provides detailed information on the test material (e.g. origin and preparation of the control and fined wines) to be used for the studies, which would include DBPCFC and skin tests but no information is given on the design of the clinical studies and on the conditions under which they will be performed. The applicant refers to a network of collaboration with clinicians that has been set up within the on-going EU funded research project REDALL (Reduced Allergenicity of processed food) and expects that this project will provide useful additional information. CONCLUSIONS AND RECOMMENDATIONS Page 6 of 7

7 The scientific data submitted by the applicant do not allow the Panel to predict the likelihood of adverse reactions in milk, egg and fish allergic individuals. Appropriate analytical and clinical studies applying good clinical and good laboratory practice should be carried out. DOCUMENTATION PROVIDED TO EFSA Dossier submitted by Deutscher Weinbauverband E.V. (DWV) to the European Commission pursuant to Article 6 paragraph 11 of Directive 2000/13/EC as amended by Directive 2003/89/EC, on 17 August REFERENCES NDA (Scientific Panel on Dietetic Products, Nutrition and Allergies) (2004). 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, PANEL MEMBERS Wulf Becker, Francesco Branca, Daniel Brasseur, Jean-Louis Bresson, Albert Flynn, Alan A. Jackson, Pagona Lagiou, Martinus Løvik, Geltrude Mingrone, Bevan Moseley, Andreu Palou, Hildegard Przyrembel, Seppo Salminen, Stephan Strobel, Henk van den Berg, and Hendrik van Loveren. ACKNOWLEDGEMENT The Scientific Panel on Dietetic Products, Nutrition and Allergies wishes to thank Elide Pastorello, Martin Stern, and Jean-Michel Wal for their contributions to the draft opinion. Page 7 of 7