Determination of volatile ‘restricted substances’ in flavourings and their volatile raw materials by GC-MS T. Cachet,* H. Brevard, E. Cantergiani, A. Chaintreau, J. Demyttenaere, L. French, K. Gassenmeier, D. Joulain, T. Koenig, H. Leijs, P. Liddle, G. Loesing, M. Marchant, K. Saito, F. Scanlan, C. Schippa, A. Scotti, F. Sekiya, A. Sherlock and T. Smith ABSTRACT: Many flavour regulations around the world contain a list of so-called ‘restricted substances’ (RS), i.e. substances that occur naturally in source materials for flavourings and food ingredients with flavouring properties, but whose presence in certain foods is restricted and/or for which maximum levels are set, for example, the European regulation 1334/2008. Only a few publications refer to the determination of RS in compound flavourings or their raw materials, and the latter only concern the analysis of one or two individual RS in single essential oils. The Working Group on Methods of Analysis of the International Organization of the Flavor Industry (IOFI) has developed a method for the rapid routine determination of β-asarone, coumarin, menthofuran, methylchavicol, methyleugenol, pulegone, safrole and α- and β-thujones in flavourings and their raw materials by gas chromatography-mass spectrometry (GC-MS), using selected-ion monitoring and internal stan- dards. The method has been evaluated by nine flavour-industry laboratories using a complex surrogate flavouring containing all of the above analytes, at concentrations that would be likely to produce levels in finished foods of around typical maximum limits for these RS. Results were obtained from a total of 15 columns and sets of analytical conditions, using 11 GC/MS instruments, with in each case a determination of the analyte in two versions of the flavouring. With reproducibility relative standard deviations (RSD R ) of less than about 20%, and recoveries of 80–120%, the method performance can be considered as satisfactory for rapid routine checks on the levels of restricted substances in compound flavourings. The method is intended for flavour-industry laboratories in order for them to fulfil their obligation to inform food-industry clients of the amounts of these substances in commercial flavourings, but is not intended for their analysis in finished foods. Copyright © 2014 John Wiley & Sons, Ltd. Keywords: gas chromatography; mass spectrometry; selected-ion monitoring; flavourings; restricted substances; quantitative analysis Introduction Since 1999, the European Scientific Committee on Food, and subsequently the European Food Safety Authority (EFSA), has expressed opinions on a number of substances occurring natu- rally in source materials for flavourings and food ingredients with flavouring properties, e.g. herbs and spices, which raise tox- icological concern. Annex III, Part B, of the European Regulation 1334/2008 [1] specifies maximum levels for what are referred to as ‘ certain substances’, together with the ‘compound food in which the presence of this substance is restricted’. Although these are sometimes referred to within the food industry as ‘Biologi- cally Active Principles’ (so-called ‘BAPs’) they will in this method be referred to as ‘Restricted Substances’ (RS). Similar limits exist for some or all of these substances in other national legislations around the world, whereas the Codex Alimentarius Commission refers to ‘Flavouring substances and components of natural flavouring complexes that may require some risk management measures’ in the Guidelines on the Use of Flavourings [2] without indicating specific substances and limits, which a previous Codex document did. Although the lat- ter General Requirements for Natural Flavourings [3] containing a list of RS has been revoked, the list, or modifications of it, still exist in many flavour regulations around the world. For example a list of substances to be controlled can be found in the Mercosur Technical Regulation Concerning Flavourings, [4] the new Russian Federation Customs Union Technical Regulation on Food Additives [5] and many flavour regulations of South-East Asian countries, such as Malaysia, Indonesia and Singapore. Many published methods exist for the determination of RS in finished foods and beverages, of which a selection is given in the References section, [6–19] but there are only a few that refer to their determination in compound flavourings or their raw mate- rials. [20–24] The latter mainly concern the analysis in single essen- tial oils of one or two individual RS, and are often based on a direct gas chromatography (GC) or liquid chromatography (LC) analysis. * Correspondence to: T. Cachet, IOFI (International Organization of the Flavor Industry) Working Group on Methods of Analysis. E-mail: tcachet@iofiorg.org IOFI (International Organization of the Flavor Industry), Working Group on Methods of Analysis Flavour Fragr. J. 2015, 30, 160–164 Copyright © 2014 John Wiley & Sons, Ltd. Research Article Received: 24 March 2014, Revised: 10 September 2014, Accepted: 17 September 2014 Published online in Wiley Online Library: 10 December 2014 (wileyonlinelibrary.com) DOI 10.1002/ffj.3222 160