Flavouring Group Evaluation 35, (FGE.35) 1 : Three quinine salts from the Priority list from chemical group 30

The EFSA Journal (2008) 739, 118 Flavouring Group Evaluation 35, (FGE.35) 1 : Three quinine salts from the Priority list from chemical group 30 Opinion of the Scientific Panel on Food Additives, Flavourings, Processing Aids and Materials in Contact with Food (EFSAQ2003172B) Adopted on 22 May 2008 SUMMARY The Scientific Panel on Food Additives, Flavourings, Processing Aids and Materials in Contact with Food (the Panel) is asked to advise the Commission on the implications for human health of chemically defined flavouring substances used in or on foodstuffs in the Member States. In particular, the Scientific Panel is asked to evaluate three flavouring substances in the Flavouring Group Evaluation 35 (FGE.35) using the Procedure as referred to in the Commission Regulation (EC) No 1565/2000. These three flavouring substances belong to chemical group 30, Annex I of the Commission Regulation (EC) No 1565/2000. Quinine sulphate [FLno: 14.152] and quinine monohydrochloride dihydrate [FLno: 14.155] are temporarily accepted as flavouring substance, while awaiting reevaluation by EFSA (compare Annex II). The Panel is aware that existing evaluations of quinine were finished more than 15 years ago and that more recent warnings have been issued about persons who may be particularily sensitive to quinine (see Annex II). The Panel therefore recommends that the toxicological database on quinine should be reconsidered. For quinine monohydrochloride dihydrate [FLno: 14.155] the estimated intake, based on the Theoretical Added Maximum Daily Intake (mtamdi) approach, is 33000 μg/person/day (corresponding to 27000 μg/person/day for the quinine base), which exceeds the threshold of concern of 90 microgram/person/day for a structural class III compound. Therefore, for this substance more reliable exposure data are required in order to finalise the evaluation. 1 For citation purposes: Scientific Opinion of the Panel on Food Additives, Flavourings, Processing Aids and Materials in Contact with Food on a request from Commission on Flavouring Group Evaluation 35, (FGE.35) Three quinine salts from the Priority list from chemical group 30. The EFSA Journal (2008) 739, 12

For quinine hydrochloride [FLno: 14.011] and quinine sulphate [FLno: 14.152] use levels are required in order to calculate the mtamdis in order to conclude whether more refined exposure assessments are needed and to finalise the evaluation. For quinine hydrochloride [FLno: 14.011] the evaluation using the Procedure cannot be finalised, as data for production volumes as flavouring substance in order to calculate an MSDI are missing. Keywords Flavourings, safety, quinine salts. The EFSA Journal (2008) 739, 217

TABLE OF CONTENTS Summary... 1 Keywords... 2 Background... 4 Terms of Reference... 6 Assessment... 6 1. Presentation of the Substances in the Flavouring Group Evaluation 35... 6 1.1. Description... 6 1.2. Stereoisomers... 6 1.3. Natural Occurrence in Food... 6 2. Specifications... 6 3. Intake Data... 7 3.1. Estimated Daily per Capita Intake (MSDI Approach)... 7 4. Evaluation / Regulatory Status... 9 5. Conclusions... 9 Table 1: Specification Summary of the Substances in the Flavouring Group Evaluation 35... 11 Annex I: Procedure for the Safety Evaluation... 12 Annex II: Evaluation/ Regulation Status... 14 References:... 16 Scientific Panel Members... 18 Acknowledgement... 18 The EFSA Journal (2008) 739, 317

BACKGROUND Regulation (EC) No 2232/96 of the European Parliament and the Council (EC, 1996) lays down a Procedure for the establishment of a list of flavouring substances, the use of which will be authorised to the exclusion of all other substances in the EU. In application of that Regulation, a Register of flavouring substances used in or on foodstuffs in the Member States was adopted by Commission Decision 1999/217/EC (EC, 1999a), as last amended by Commission Decision 2008/478/EC (EC, 2008). Each flavouring substance is attributed a FLAVISnumber (FLnumber) and all substances are divided into 34 chemical groups. Substances within a group should have some metabolic and biological behaviour in common. Substances which are listed in the Register are to be evaluated according to the evaluation programme laid down in Commission Regulation (EC) No 1565/2000 (EC, 2000a), which is broadly based on the Opinion of the Scientific Committee on Food (SCF, 1999). For the submission of data by the manufacturer, deadlines have been established by Commission Regulation (EC) No 622/2002 (EC, 2002b). After the completion of the evaluation programme the positive list of flavouring substances for use in or on foods in the EU shall be adopted (Article 5 (1) of Regulation (EC) No 2232/96) (EC, 1996). By Commission Decision 1999/217/EC certain flavouring substances received priority in the evaluation programme, since concerns about the safety of consumers were expressed by some Member States. In the Register, these substances received the following remark: 3. Substance to be given priority evaluation. In the Commission Decision 1999/217/EC, Part 1 to 3, 18 flavouring substances received this remark. In May 2000, a revised list containing 49 flavouring substances with this remark was adopted (Commission Decision 2000/489/EC amending Commission Decision 1999/217/EC). By the amendment from January 2002 (Commission Decision 2002/113/EC amending Commission Decision 1999/217/EC) the list contains 43 flavouring substances with this remark (see Table A). Table A. Status of 43 Flavouring Substances Given Priority Evaluation (in Commission Decision 2002/113/EC amending Commission Decision 1999/217/EC) FLno Name Status in the FLAVIS database, May 2008 01.015 Vinylbenzene FGE.29 Priority substance. 08.017 Malic acid JECFA no: 619. 08.041 Octadeca9,12dienoic acid JECFA no: 332. 13.018 Furfural EFSA Opinion adopted 2004. ADI of 0.5 mg/kg bw. FGE.19 EFSA evaluation of alpha,betaunsaturated aldehydes and ketones under evaluation. 13.035 Menthofuran FGE.57 EFSA consideration of JECFA evaluated substances under evaluation. 13.126 Furfural diethyl acetal EFSA Opinion adopted 2004. ADI of 0.5 mg/kg bw. FGE.19 EFSA evaluation of alpha,betaunsaturated aldehydes and ketones under evaluation. 14.011 Quinine hydrochloride FGE.35 JECFA evaluation 1993: No concern up to 100 mg/l in soft drink (Commission Directive 2002/67/EC) (EC, 2002c). 14.152 Quinine sulphate FGE.35 JECFA evaluation 1993: No concern up to 100 mg/l in soft drink (Commission Directive 2002/67/EC) (EC, 2002c). 14.155 Quinine monohydrochloride dihydrate FGE.35 JECFA evaluation 1993: No concern up to 100 mg/l in soft drink (Commission Directive 2002/67/EC) (EC, 2002c). 16.002 Diammonium sulfide Not allocated any evaluation suggested for deletion by DG SANCO. 16.012 Glycyrrhizic acid FGE.36 SCF Opinion adopted 2003 (Commission Directive 2004/77/EC) (EC, 2004b). JECFA evaluation 2004: Data were inadequate to derive an ADI. The EFSA Journal (2008) 739, 417

Table A. Status of 43 Flavouring Substances Given Priority Evaluation (in Commission Decision 2002/113/EC amending Commission Decision 1999/217/EC) FLno Name Status in the FLAVIS database, May 2008 16.016 Caffeine FGE.49 SCF Opinions adopted 1999, 2003. (Commission Directive 2002/67/EC) (EC, 2002c). 16.027 Thiamine hydrochloride FGE.76 Adopted EFSA consideration of JECFA evaluated substances. 16.032 Theobromine FGE.49 SCF Opinions adopted 1999, 2003 on caffeine. 16.056 Taurine FGE.79 Adopted EFSA consideration of JECFA evaluated substances. 16.059 Ammonium hydrogen sulphide FGE.46 EFSA evaluation of ammonium and ammonium salts under evaluation. 16.060 Glycyrrhizic acid, ammoniated FGE.36 SCF Opinion adopted 2003 (Commission Directive 2004/77/EC) (EC, 2004b). JECFA evaluation 2004: Data were inadequate to derive an ADI. 17.001 betaalanine FGE.79 Adopted EFSA consideration of JECFA evaluated substances. 17.002 lalanine FGE.26Rev1 Adopted EFSA evaluation of amino acids. 17.003 Arginine FGE.79 Adopted EFSA consideration of JECFA evaluated substances. 17.005 Aspartic acid FGE.79 Adopted EFSA consideration of JECFA evaluated substances. 17.006 Cystine FGE.26 Adopted EFSA evaluation of amino acids. 17.007 Glutamine FGE.79 Adopted EFSA consideration of JECFA evaluated substances. 17.008 Histidine FGE.79 Adopted EFSA consideration of JECFA evaluated substances. 17.010 Isoleucine FGE.79 Adopted EFSA consideration of JECFA evaluated substances. 17.012 Leucine FGE.79 Adopted EFSA consideration of JECFA evaluated substances. 17.013 Lysine FGE.26 Adopted EFSA evaluation of amino acids. 17.014 Methionine FGE.79 Adopted EFSA consideration of JECFA evaluated substances. 17.017 Phenylalanine (DL) FGE.79 Adopted EFSA consideration of JECFA evaluated substances. 17.018 Phenylalanine (L) FGE.79 Adopted EFSA consideration of JECFA evaluated substances. 17.019 Proline FGE.79 Adopted EFSA consideration of JECFA evaluated substances. 17.020 Serine FGE.26 Adopted EFSA evaluation of amino acids. 17.021 Threonine FGE.26 Adopted EFSA evaluation of amino acids. 17.022 Thyrosine FGE.79 Adopted EFSA consideration of JECFA evaluated substances. 17.023 Valine FGE.79 Adopted EFSA consideration of JECFA evaluated substances. 17.024 Alanine FGE.79 Adopted EFSA consideration of JECFA evaluated substances. 17.026 Lysine FGE.79 Adopted EFSA consideration of JECFA evaluated substances. 17.027 Methionine FGE.26Rev1 Adopted EFSA evaluation of amino acids. 17.028 Valine FGE.26Rev1 Adopted EFSA evaluation of amino acids. 17.031 Lysine monochlorhydrate FGE.26 Adopted EFSA evaluation of amino acids. 17.032 Cysteine hydrochloride FGE.26Rev1 Adopted EFSA evaluation of amino acids. 17.033 Cysteine FGE.79 Adopted EFSA consideration of JECFA evaluated substances. 17.034 Glycine FGE.79 Adopted EFSA consideration of JECFA evaluated substances. Of the 43 substances in Table A, one is suggested for deletion from the Register, 12 have been evaluated in other FGEs, and 22 have been evaluated by JECFA (20 of these JECFA evaluated substances are also considered by EFSA). The remaining eight substances, vinylbenzene [FLno: 01.015], quinine hydrochloride [FLno: 14.011], quinine sulphate [FLno: 14.152], quinine monohydrochloride dihydrate [FLno: 14.155], glycyrrhizic acid [FLno: 16.012], glycyrrhizic acid, ammoniated [FLno: 16.060], caffeine [FLno: 16.016] and theobromine [FLno: 16.032] are considered in the following Flavouring Group Evaluations: FGE.29: Aromatic hydrocarbon: vinylbenzene [FLno: 01.015]. FGE.35: Quinoline alkaloids: quinine hydrochloride [FLno: 14.011], quinine sulphate [FLno: 14.152], quinine monohydrochloride dihydrate [FLno: 14.155]. The EFSA Journal (2008) 739, 517

FGE.36: Triterpene glycosides: glycyrrhizic acid [FLno: 16.012], glycyrrhizic acid, ammoniated [FLno: 16.060]. FGE.49: Xanthin alkaloids: caffeine [FLno: 16.016] and theobromine [FLno: 16.032]. TERMS OF REFERENCE European Food Safety Authority (EFSA) is requested to carry out a risk assessment on flavouring substances prior to their authorisation and inclusion in a positive list according to Commission Regulation (EC) No 1565/2000 (EC, 2000a). ASSESSMENT 1. Presentation of the Substances in the Flavouring Group Evaluation 35 1.1. Description The present, using the Procedure as referred to in the Commission Regulation (EC) No 1565/2000 (EC, 2000) (The Procedure shown in schematic form in Annex I), deals with three quinine alkaloids, quinine hydrochloride [FLno: 14.011], quinine sulphate [FLno: 14.152] and quinine monohydrochloride dihydrate [FLno: 14.155] from EU chemical group 30, Annex I of Commission Regulation (EC) No 1565/2000 (EC, 2000). The three flavouring substances under consideration, as well as their chemical Register names, FLAVIS (FL), Chemical Abstract Service (CAS), Council of Europe (CoE) and Flavor and Extract Manufacturers Association (FEMA) numbers, structure and specifications, are listed in Table 1. 1.2. Stereoisomers It is recognised that geometrical and optical isomers of substances may have different properties. Their flavour may be different, they may have different chemical properties resulting in possible variation of their absorption, distribution, metabolism, elimination and toxicity. Thus, information must be provided on the configuration of the flavouring substance, i.e. whether it is one of the geometrical/optical isomers, or a defined mixture of stereoisomers. The available specifications of purity will be considered in order to determine whether the safety evaluation carried out for candidate substances for which stereoisomers may exist can be applied to the material of commerce. Flavouring substances with different configurations should have individual chemical names and codes (CAS number, FLAVIS number, etc.). The configurations of the three flavouring substances in this FGE are determined by their chemical names: quinine refers to the ()isomer, which has (8S,9R)configuration. 1.3. Natural Occurrence in Food Quinine does not occur naturally in food. 2. Specifications Purity criteria for the three substances have been provided by the Flavour Industry (Table 1). Judged against the requirements in Annex II of Commission Regulation (EC) No 1565/2000 (EC, 2000), the information is adequate for the three candidate substances except that the purity criteria The EFSA Journal (2008) 739, 617

for quinine sulphate [FLno: 14.152] is deficient in one of the parameters as no minimum assay is provided and for quinine monohydrochloride dihydrate [FLno: 14.155] the composition of the secondary components are to be specified (see Section 1.2 and Table 1). 3. Intake Data Annual production volumes of the flavouring substances as surveyed by the Industry can be used to calculate the Maximised SurveyDerived Daily Intake (MSDI) by assuming that the production figure only represents 60 % of the use in food due to underreporting and that 10 % of the total EU population are consumers (SCF, 1999). However, the Panel noted that due to yeartoyear variability in production volumes, to uncertainties in the underreporting correction factor and to uncertainties in the percentage of consumers, the reliability of intake estimates on the basis of the MSDI approach is difficult to assess. The Panel also noted that in contrast to the generally low per capita intake figures estimated on the basis of this MSDI approach, in some cases the regular consumption of products flavoured at use levels reported by the Flavour Industry in the submissions would result in much higher intakes. In such cases, the human exposure thresholds below which exposures are not considered to present a safety concern might be exceeded. Considering that the MSDI model may underestimate the intake of flavouring substances by certain groups of consumers, the SCF recommended also taking into account the results of other intake assessments (SCF, 1999). One of the alternatives is the Theoretical Added Maximum Daily Intake (TAMDI) approach, which is calculated on the basis of standard portions and upper use levels (SCF, 1995) for flavourable beverages and foods in general, with exceptional levels for particular foods. This method is regarded as a conservative estimate of the actual intake in most consumers because it is based on the assumption that the consumer regularly eats and drinks several food products containing the same flavouring substance at the upper use level. One option to modify the TAMDI approach is to base the calculation on normal rather than upper use levels of the flavouring substances. This modified approach is less conservative (i.e. it may underestimate the intake of consumers being loyal to products flavoured at the maximum use levels reported (EC, 2000a). However, it is considered as a suitable tool to screen and prioritise the flavouring substances according to the need for refined intake data (EFSA, 2004a). 3.1. Estimated Daily per Capita Intake (MSDI Approach) The Maximised SurveyDerived Daily Intake (MSDI (SCF, 1999)) data are derived from surveys on annual production volumes in Europe. These surveys were conducted in 1995 by the International Organization of the Flavour Industry, in which flavour manufacturers reported the total amount of each flavouring substance incorporated into food sold in the EU during the previous year (IOFI, 1995). The intake approach does not consider the possible natural occurrence in food. The EFSA Journal (2008) 739, 717

Average per capita intake (MSDI) is estimated on the assumption that the amount added to food is consumed by 10 % of the population 2 (Eurostat, 1998). This is derived for candidate substances from estimates of annual volume of production provided by Industry and incorporates a correction factor of 0.6 to allow for incomplete reporting (60 %) in the Industry surveys (SCF, 1999). The MSDI and mtamdi values for quinine hydrochloride [FLno: 14.011], quinine sulphate [FLno: 14.152] and quinine monohydrochloride dihydrate [FLno: 14.155] are shown in Table 3.1. Table 3.1 Estimated intakes based on the MSDI approach and the mtamdi approach FLno EU Register name MSDI (μg/capita/day) mtamdi (μg/person/day) Structural class Threshold of concern (µg/person/day) 14.011 Quinine hydrochloride Class III 90 14.152 Quinine sulphate 0.12 Class III 90 14.155 Quinine monohydrochloride dihydrate 120 33000 Class III 90 For quinine sulphate [FLno: 14.152] and quinine monohydrochloride dihydrate [FLno: 14.155] the MSDI values are 0.12 and 120 μg/capita/day, respectively, corresponding to 0.11 and 98 μg/capita/day of the quinine base. Table 3.2 shows the normal and maximum use levels for the candidate substance [FLno: 14.155]. The Industry submission does not indicate whether the use levels are maximum or normal use levels. Normal use is defined as the average of reported usages and maximum use is defined as the 95 th percentile of reported usages (EFFA, 2002i). Table 3.2 Normal and Maximum use levels (mg/kg) for the substances in FGE.35 FLno Food Categories Normal use levels (mg/kg) Maximum use levels (mg/kg) 01.0 02.0 03.0 04.1 04.2 05.0 06.0 07.0 08.0 09.0 10.0 11.0 12.0 13.0 14.1 14.2 15.0 16.0 14.155 25 15 85 100 Table 3.3 Food categories according to Commission Regulation (EC) No 1565/2000 (EC, 2000) Food category Description 01.0 Dairy products, excluding products of category 02.0 02.0 Fats and oils, and fat emulsions (type waterinoil) 03.0 Edible ices, including sherbet and sorbet 04.1 Processed fruit 04.2 Processed vegetables (incl. mushrooms & fungi, roots & tubers, pulses and legumes), and nuts & seeds 05.0 Confectionery 06.0 Cereals and cereal products, incl. flours & starches from roots & tubers, pulses & legumes, excluding bakery 07.0 Bakery wares 08.0 Meat and meat products, including poultry and game 09.0 Fish and fish products, including molluscs, crustaceans and echinoderms 10.0 Eggs and egg products 11.0 Sweeteners, including honey 12.0 Salts, spices, soups, sauces, salads, protein products, etc. 13.0 Foodstuffs intended for particular nutritional uses 14.1 Nonalcoholic ("soft") beverages, excl. dairy products 14.2 Alcoholic beverages, incl. alcoholfree and lowalcoholic counterparts 2 EU figure 375 millions (Eurostat, 1998). This figure relates to EU population at the time for which production data are available, and is consistent (comparable) with evaluations conducted prior to the enlargement of the EU. No production data are available for the enlarged EU. The EFSA Journal (2008) 739, 817

Table 3.3 Food categories according to Commission Regulation (EC) No 1565/2000 (EC, 2000) Food category Description 15.0 Readytoeat savouries 16.0 Composite foods (e.g. casseroles, meat pies, mincemeat) foods that could not be placed in categories 01.0 15.0 4. Evaluation / Regulatory Status Quinine hydrochloride [FLno: 14.011], quinine sulphate [FLno: 14.152] and quinine monohydrochloride dihydrate [FLno: 14.155] have been previously evaluated by the Scientific Committee for Food in relation to investigate the safetyinuse of quinine in food and drinks: Some individuals are hypersensitive to quinine, as occurs with other food components and food additives. These persons should be informed by the specific mention of the presence of quinine on the label. The Committee sees no objections from a toxicological point of view to the continued use at present levels (up to max. 100 mg/l) of quinine in bitter drinks (SCF, 1988). Quinine hydrochloride has been evaluated by the JECFA at its 39 th meeting: The Committee concluded that current levels of use up to 100 mg/l (as quinine base) in soft drinks were not of toxicological concern. The contribution of other uses of quinine in food and alcoholic beverages to daily intakes was considered to be negligible (JECFA, 1992a) and again at its 41 st meeting (JECFA, 1993a). An amendment to the EC labelling Directive (European Council Directive 2000/13/EC) requires that, by derogation from Article 6(6), second subparagraph, third indent, of Directive 2000/13/EC, quinine and/or caffeine used as a flavouring in the production or preparation of a foodstuff must be mentioned by name in the list of ingredients indicated in Article 3(1)(2), of Directive 2000/13/EC, immediately after the term flavouring (EC, 2000b). The German Federal Institute for Risk Assessment has recently published an updated health assessment: Quininecontaining beverages may cause health problems (BfR, 2008). Annex II provides more detail on the outcome of these evaluations. 5. Conclusions The Panel is aware that existing evaluations of quinine were finished more than 15 years ago when the SCF concluded that Some individuals are hypersensitive to quinine, as occurs with other food components and food additives. These persons should be informed by the specific mention of the presence of quinine on the label and that The Committee sees no objections from a toxicological point of view to the continued use at present levels (up to max. 100 mg/l) of quinine in bitter drinks. The Panel is also aware that more recent warnings have been issued about persons who may be particularily sensitive to quinine (see Annex II). The Panel therefore recommends that the toxicological database on quinine should be reconsidered. For quinine monohydrochloride dihydrate [FLno: 14.155] the estimated intake, based on the mtamdi approach is 33000 μg/person/day (corresponding to 27000 μg/person/day for the quinine base), which exceeds the threshold of concern of 90 microgram/person/day for a structural class III compound. Therefore, for this substance more reliable exposure data are required in order to finalise the evaluation. For quinine hydrochloride [FLno: 14.011] and quinine sulphate [FLno: 14.152] use levels are required in order to calculate the mtamdis in order to conclude whether more refined exposure assessments are needed and to finalise the evaluation. The EFSA Journal (2008) 739, 917

For quinine hydrochloride [FLno: 14.011] the evaluation using the Procedure cannot be finalised, as data for production volumes as flavouring substance in order to calculate an MSDI are missing. In order to determine whether the conclusion for the candidate substances can be applied to the materials of commerce, it is necessary to consider the available specifications: Specifications for the three candidate substances have been provided. Information on minimum assay is missing for quinine sulphate [FLno: 14.152] and for quinine monohydrochloride dihydrate [FLno: 14.155] the composition of the secondary components are to be specified. The EFSA Journal (2008) 739, 1017

TABLE 1: SPECIFICATION SUMMARY OF THE SUBSTANCES IN THE FLAVOURING GROUP EVALUATION 35 Table 1: Specification Summary of the Substances in the Flavouring Group Evaluation 35 FLno EU Register name Structural formula FEMA no CoE no CAS no 14.011 Quinine hydrochloride N O OH N HCl 2976 715 130892 Phys. form Mol. formula Mol. weight Solubility 1) Solubility in ethanol 2) Boiling point, C 3) Melting point, C ID test Assay minimum Solid Soluble C 20H 24N 2O 2 HCl Freely soluble 115116 396.91 NMR MS 95 % Refrac. Index 4) Spec. gravity 5) n.a. n.a. Specification comments 14.152 Quinine sulphate N O OH N H 2SO 4 2977 804637 Solid (C 20H 24N 2O 2) 2 H 2SO 4 2H 2O 782.96 Soluble Soluble 225 IR n.a. n.a. AV 6). 14.155 Quinine monohydrochloride dihydrate N O OH N HCl. 2H 2O 6119477 Solid Soluble C 20H 24N 2O 2 HCl Very soluble 115116 2H 2O 396.92 NMR 90 % n.a. n.a. Minimum assay is "90 %". Composition of secondary components to be specified. 1) Solubility in water, if not otherwise stated. 2) Solubility in 95 % ethanol, if not otherwise stated. 3) At 1013.25 hpa, if not otherwise stated. 4) At 20 C, if not otherwise stated. 5) At 25 C, if not otherwise stated. 6) AV: Missing minimum assay value. The EFSA Journal (2008) 739, 1117

ANNEX I: PROCEDURE FOR THE SAFETY EVALUATION The approach for a safety evaluation of chemically defined flavouring substances as referred to in Commission Regulation (EC) No 1565/2000 (EC, 2000a), named the "Procedure", is shown in schematic form in Figure I.1. The Procedure is based on the Opinion of the Scientific Committee on Food expressed on 2 December 1999 (SCF, 1999), which is derived from the evaluation procedure developed by the Joint FAO/WHO Expert Committee on Food Additives at its 44 th, 46 th and 49 th meetings (JECFA, 1995; JECFA, 1996a; JECFA, 1997a; JECFA, 1999b). The Procedure is a stepwise approach that integrates information on intake from current uses, structureactivity relationships, metabolism and, when needed, toxicity. One of the key elements in the Procedure is the subdivision of flavourings into three structural classes (I, II, III) for which thresholds of concern (human exposure thresholds) have been specified. Exposures below these thresholds are not considered to present a safety concern. Class I contains flavourings that have simple chemical structures and efficient modes of metabolism, which would suggest a low order of oral toxicity. Class II contains flavourings that have structural features that are less innocuous, but are not suggestive of toxicity. Class III comprises flavourings that have structural features that permit no strong initial presumption of safety, or may even suggest significant toxicity (Cramer et al., 1978). The thresholds of concern for these structural classes of 1800, 540 or 90 microgram/person/day, respectively, are derived from a large database containing data on subchronic and chronic animal studies (JECFA, 1996a). In Step 1 of the Procedure, the flavourings are assigned to one of the structural classes. The further steps address the following questions: can the flavourings be predicted to be metabolised to innocuous products 3 (Step 2)? do their exposures exceed the threshold of concern for the structural class (Step A3 and B3)? are the flavourings or their metabolites endogenous 4 (Step A4)? does a NOAEL exist on the flavourings or on structurally related substances (Step A5 and B4)? In addition to the data provided for the flavouring substances to be evaluated (candidate substances), toxicological background information available for compounds structurally related to the candidate substances is considered (supporting substances), in order to assure that these data are consistent with the results obtained after application of the Procedure. The Procedure is not to be applied to flavourings with existing unresolved problems of toxicity. Therefore, the right is reserved to use alternative approaches if data on specific flavourings warranted such actions. 3 Innocuous metabolic products : Products that are known or readily predicted to be harmless to humans at the estimated intakes of the flavouring agent (JECFA, 1997a). 4 Endogenous substances : Intermediary metabolites normally present in human tissues and fluids, whether free or conjugated; hormones and other substances with biochemical or physiological regulatory functions are not included (JECFA, 1997a). The EFSA Journal (2008) 739, 1217

Procedure for Safety Evaluation of Chemically Defined Flavouring Substances Step 1. Decision tree structural class Step 2. Can the substance be predicted to be metabolised to innocuous products? Step A3. Yes Step B3. No Do the conditions of use result in an intake greater than the threshold of concern for the structural class? Data must be available on the substance or closely related substances to perform a safety evaluation Yes Do the conditions of use result in an intake greater than the threshold of concern for the structural class? Step A4. Yes No Step B4. No Is the substanc e or are its metabolites endogenous? Yes Substance would not be expected to be of safety concern Yes Does a NOAEL exist for the substance which provides an adequate margin of safety under conditions of intended use, or does a NOAEL exist for structurally related substances which is high enough to acc ommodate any perc eived difference in toxicity between the substance and the related substances? Step A5. No Yes No Does a NOAEL exist for the substance which provides an adequate margin of safety under conditions of intended use, or does a NOAEL exist for structurally related substances which is high enough to accommodate any perceived difference in toxicity between the substance and the related substances? No Additional data required Figure I.1 Procedure for Safety Evaluation of Chemically Defined Flavouring Substances The EFSA Journal (2008) 739, 1317

ANNEX II: EVALUATION/ REGULATION STATUS Quinine hydrochloride [FLno: 14.011], quinine sulphate [FLno: 14.152] and quinine monohydrochloride dihydrate [FLno: 14.155] SCF report of the Scientific Committee for food on quinine (Opinion expressed 19 February 1988) The SCF acknowledges that it has been provided with comprehensive additional information and new studies in response to its requests. The Committee is now assured that no adverse reproductive or teratological effects will result from the use of quinine in bitter drinks. The Committee has also been provided with information on actual and potential intakes of quinine from bitter soft drinks at a European level. The estimated actual intake in European countries is, on average, of the order of 0.26 mg/person/day, and for regular consumers of bitter drinks it is unlikely that the mean daily intake will exceed 5 mg quinine per person/day. This information is reassuring for the Committee and it has noted that intake appears to be restricted to the adult population. Military jetpilots consuming 105 mg quinine daily showed, under extremely strenuous conditions, mild adverse effects, but these effects are not considered relevant in the context of the use of quinine as a food additive. For human volunteers under normalconditions 120 mg/person/day gave no effects. This should not be considered in relation to the estimated maximum daily intake of 5 mg/person/day in Member states. Some individuals are hypersensitive to quinine, as occurs with other food components and food additives. These persons should be informed by the specific mention of the presence of quinine on the label. The Committee sees no objections from a toxicological point of view to the continued use at present levels (up to max. 100 mg/l) of quinine in bitter drinks. JECFA evaluation of quinine (JECFA, 1993) Quinine was last reviewed by the Committee at its thirtyninth meeting, when a clear No Observed Effect Level (NOEL) with respect to ocular effects in humans of 80 mg of anhydrous quinine hydrochloride per day (equivalent to 72 mg of free base) was observed. At that time, the Committee withdrew the previously established temporary Acceptable Daily Intake (ADI) of 00.9 mg of quinine per kg body weight per day and concluded that current levels of use in soft drinks, which the Committee was informed were up to 75 mg/l (as quinine base), were not of toxicological concern. At its present meeting, the Committee reassessed the toxicological information in the light of new data on levels of quinine in beverages. The Committee concluded that current levels of use up to 100 mg/l (as quinine base) in soft drinks were not of toxicological concern. The contribution of other uses of quinine in food and alcoholic beverages to daily intakes was considered to be negligible. The Committee again noted that certain consumers show an idiosyncratic hyperreactivity to quinine, and reiterated its recommendation that the consumer should be informed by appropriate means of the presence of quinine in foods and beverages in which it is used. A toxicological monograph was not prepared. The existing specifications for quinine hydrochloride and quinine sulfate were revised. EU Regulation (Commission Directive 2002/67/EC) The EFSA Journal (2008) 739, 1417

There is an amendment (Commission Directive 2002/67/EC (EC, 2002c) to the EC labelling directive (European Council Directive 2000/13/EC): By derogation from Article 6(6), second subparagraph, third indent, of Directive 2000/13/EC, quinine and/or caffeine used as a flavouring in the production or preparation of a foodstuff must be mentioned by name in the list of ingredients indicated in Article 3(1)(2), of Directive 2000/13/EC, immediately after the term flavouring (EC, 2000b). German Federal Institute for risk Assessment (BfR): Quininecontaining beverages may cause health problems (BfR, 2008) Summary: Quinine is a bittertasting, crystalline white powder. It is obtained from the bark of the cinchona tree and belongs to the group of alkaloids. In medicine quinine is used to treat malaria and nocturnal leg cramps. In the food sector, quinine is used as a flavouring mainly in beverages like bitter lemon and tonic water. When larger amounts of quinine are consumed, it can constitute a health problem for some consumer groups. BfR sees risks in particular for quinine intakes during pregnancy. For instance, a newborn baby, whose mother had drunk more than 1 litre tonic water a day in the weeks up to its birth, suffered health disorders. Based on existing regulations in the medicinal product sector, BfR, therefore, advises pregnant women against drinking quininecontaining beverages on precautionary grounds. People who have been advised against taking quinine, cinchona bark or their preparations by their doctors because of their clinical pictures should not consume any quininecontaining soft drinks either. This applies, for instance, to people who suffer from tinnitus, preexisting damage to the optic nerve, haemolytic anaemia or who are hypersensitive to quinine or cinchona alkaloids. Patients with cardiac arrhythmia and people who take medicine that interacts with quinine, should only drink quininecontaining soft drinks after consulting their doctors. This applies in particular to medications which inhibit blood coagulation. At higher levels of tonic water consumption, it may be necessary to reduce their therapeutic dose. Already today quinine must be mentioned by name in the list of ingredients of quininecontaining products. BfR also believes that there is a need for information which attracts the attention more particularly of pregnant women and other risk groups to possible health impairments. Motor vehicle drivers should be informed that larger amounts of quininecontaining bitter beverages can cause visual disturbances. BfR recommends raising awareness about the possible health risks from quinine to consumers. Specific information should be provided about the symptoms of quinine hypersensitivity and cinchonism (typical adverse reactions to quinine). Consumers should be advised to immediately stop their quinine intake if these symptoms occur, and to consult a doctor. BfR recommends that the health assessment of quinine by the Scientific Committee on Food from 1988 should be updated. BfR is of the opinion that the problems of quininecontaining bitter soft drinks underline the importance of the systematic recording of adverse reactions that occur in conjunction with the consumption of foods. The Institute, therefore, explicitly supports the setting up of a central reporting office. The EFSA Journal (2008) 739, 1517

REFERENCES: BfR, 2008. Quininecontaining beverages may cause health problems. Federal Institute for Risk Assessment. Updated BfR Health Assessment. No 020/2008, 17 February 2005. Updated 9 May 2008. [Online] http://www.bfr.bund.de/cm/245/quinine_containing_beverages_may_cause_health_problems.pdf (as of 19 May 2008). Cramer, G.M., Ford, R.A., Hall, R.L., 1978. Estimation of toxic hazard a decision tree approach. Food Cosmet. Toxicol. 16(3), 255 276. EC, 1996. Regulation No 2232/96 of the European Parliament and of the Council of 28 October 1996. Official Journal of the European Communities 23.11.1996, L 299, 14. EC, 1999a. Commission Decision 1999/217/EC of 23 February 1999 adopting a register of flavouring substances used in or on foodstuffs. Official Journal of the European Communities 27.3.1999, L 84, 1137. EC, 2000a. Commission Regulation No 1565/2000 of 18 July 2000 laying down the measures necessary for the adoption of an evaluation programme in application of Regulation (EC) No. 2232/96. Official Journal of the European Communities 19.7.2000, L 180, 816. EC, 2000b. Commission Directive 2000/13/EC of 20 March 2000 on the approximation of the laws of the Member States relating to the labelling, presentation and advertising of foodstuffs. Official Journal of the European Union 06.5.2000, L 109, 2942. EC, 2002b. Commission Regulation No 622/2002 of 11 April 2002 establishing deadlines for the submission of information for the evaluation of chemically defined flavouring substances used in or on foodstuffs. Official Journal of the European Communities 12.4.2002, L 95, 1011. EC, 2002c. Commission Directive 2002/67/EC of 18 July 2002on the labelling of foodstuffs containing quinine, and foodstuffs containing caffeine. Official Journal of the European Union 19.7.2002, L 191, 2021. EC, 2008. Commission Decision 2008/478/EC of 17 June 2008 amending Decision 1999/217/EC as regards the register of flavouring substances used in or on foodstuffs. Official Journal of the European Union 24.6.2008, L 163, 42. EFFA, 2002i. Letter from EFFA to Dr. Joern Gry, Danish Veterinary and Food Administration. Dated 31 October 2002. Re.: Second group of questions. FLAVIS/8.26. EFSA, 2004a. Minutes of the 7th Plenary meeting of the Scientific Panel on Food Additives, Flavourings, Processing Aids and Materials in Contact with Food, Held in Brussels on 1213 July 2004. Brussels, 28 September 2004. [Online]. Available: http://www.efsa.eu.int/science/afc/afc_meetings/502_en.html Eurostat, 1998. Total population. Cited in Eurostat, 2004. The EU population, Total population. [Online]. Available: http://europa.eu.int/comm/eurostat/newcronos/queen/display.do?screen=detail&language=en&product=yes&root=yes/yearlies/c /ca/caa/caa10000. August 13, 2004. IOFI, 1995. European inquiry on volume of use. IOFI, International Organization of the Flavor Industry, 1995. JECFA, 1992a. 39. Report: Thirtyninth Meeting of the Joint FAO/WHO Expert Committee on Food Additives. Report: WHO Technical Report Series, no. 828. JECFA, 1993a. 41. Report: FortyFirst Meeting of the Joint FAO/WHO Expert Committee on Food Additives. Report: WHO Technical Report Series, no. 837. JECFA, 1995. Evaluation of certain food additives and contaminants. Fortyfourth Meeting of the Joint FAO/WHO Expert Committee on Food Additives. WHO Technical Report Series, no. 859. Geneva. JECFA, 1996a. Toxicological evaluation of certain food additives. The fortyfourth meeting of the Joint FAO/WHO Expert Committee on Food Additives and contaminants. WHO Food Additives Series: 35. IPCS, WHO, Geneva. JECFA, 1997a. Evaluation of certain food additives and contaminants. Fortysixth report of the Joint FAO/WHO Expert Committee on Food Additives. Geneva, 615 February 1996. WHO Technical Report Series, no. 868. Geneva. JECFA, 1999b. Evaluation of certain food additives and contaminants. Fortyninth report of the Joint FAO/WHO Expert Committee on Food Additives. Rome, 1726 June 1997. WHO Technical Report Series, no. 884. Geneva. SCF, 1988. Report of the Scientific Committee for Food on Quinine. In food Science and techniques. (Opinion expressed on 11 November 1988). Reports of the Scientific Committee for Food (Twentyfirst series). Internal Market and industrial Affairs. DirectorateGeneral. The EFSA Journal (2008) 739, 1617

SCF, 1995. Scientific Committee for Food. First annual report on chemically defined flavouring substances. May 1995, 2 nd draft prepared by the SCF Working Group on Flavouring Substances (Submitted by the SCF Secretariat, 17 May 1995). CS/FLAV/FL/140Rev2. Annex 6 to Document III/5611/95, European Commission, DirectorateGeneral III, Industry. SCF, 1999. Opinion on a programme for the evaluation of flavouring substances (expressed on 2 December 1999). Scientific Committee on Food. SCF/CS/FLAV/TASK/11 Final 6/12/1999. Annex I the minutes of the 119 th Plenary meeting. European Commission, Health & Consumer Protection DirectorateGeneral. The EFSA Journal (2008) 739, 1717

SCIENTIFIC PANEL MEMBERS Fernando Aguilar, Herman Nybro Autrup, Susan Barlow, Laurence Castle, Riccardo Crebelli, Wolfgang Dekant, KarlHeinz Engel, Nathalie Gontard, David Michael Gott, Sandro Grilli, Rainer Gürtler, John Christian Larsen, JeanCharles Leblanc, Catherine Leclercq, F. Xavier Malcata, Wim Mennes, Maria Rosaria Milana, Iona Pratt, Ivonne Magdalena Catharina Maria Rietjens, Paul P. Tobback, Fidel Toldrá. ACKNOWLEDGEMENT The Scientific Panel on Food Additives, Flavourings, Processing Aids and Materials in Contact with Food wishes to thank Jørn Gry, Vibe Beltoft, Pia Lund and Karin Nørby for their contribution to the draft Opinion. The EFSA Journal (2008) 739, 1817