Flavouring Group Evaluation 36, (FGE.36) 1 Two triterpene glycosides from the priority list

The EFSA Journal (2008) 740, 219 Flavouring Group Evaluation 36, (FGE.36) 1 Two triterpene glycosides from the priority list pinion of the Scientific Panel on Food Additives, Flavourings, Processing Aids and Materials in Contact with Food (AFC) (EFSAQ2003172C) Adoption date 22 May 2008 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. 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 two flavouring substances in the Flavouring Group Evaluation FGE.36 (FGE.36) using the Procedure as referred to in the Commission Regulation (EC) No 1565/2000. These two flavouring substances belong to chemical group 30, Annex I of the Commission Regulation (EC) No 1565/2000. The present Flavouring Group Evaluation 36 (FGE.36) deals with two triterpene glycosides, glycyrrhizic acid [FLno: 16.012] and glycyrrhizic acid, ammoniated [FLno: 16.060] from chemical group 30. The Panel agrees with the evaluation by the Scientific Committee on Food (SCF) and emphasises the need for further information on use levels and on intakes that were requested by the SCF in its pinion from 2003 (see Annex II). The mtamdi of 210 mg/person/day exceeds by a factor of two 1 For citation purposes: Scientific pinion of the Panel on Food Additives, Flavourings, Processing Aids and Materials in Contact with Food (AFC) on a request from the Commission on Flavouring Group Evaluation 36, (FGE.36) Two triterpene glycosides from the priority list. The EFSA Journal (2008) 740, 119. European Food Safety Authority, 2008

the intake of 100 mg/person/day considered by the SCF to be a sufficient level of protection. Therefore, refined use levels and intake data are needed. Furthermore, the two substances have been presented without specification of the stereoisomeric composition. KEYWRDS Flavourings, safety, glycyrrhizic acid, ammoniated glycyrrhizic acid. The EFSA Journal (2008) 740, 319

TABLE F CNTENTS Panel Members... 2 Summary... 2 Keywords... 3 Background... 5 Acknowledgement... 7 Terms of Reference... 7 Assessment... 8 1. Presentation of the Substances in the Flavouring Group Evaluation 36... 8 1.1. Description... 8 1.2. Stereoisomers... 8 1.3. Natural ccurrence in Food... 8 2. Specifications... 8 3. Intake Data... 9 3.1. Estimated Daily per Capita Intake (MSDI Approach)... 9 4. Evaluation/ Regulatory Status... 11 5. Conclusions... 11 Table 1: Specification Summary of the Substances in the Flavouring Group Evaluation 36... 12 Annex I: Procedure for the Safety Evaluation... 14 Annex II: Evaluation/ Regulation Status... 17 References:... 19 The EFSA Journal (2008) 740, 419

BACKGRUND 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 pinion 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 the health 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 ctadeca9,12dienoic acid JECFA no: 332. 13.018 Furfural EFSA pinion 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 pinion 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)). The EFSA Journal (2008) 740, 519

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 16.002 Diammonium sulfide Not allocated any evaluation suggested for deletion by DG SANC. 16.012 Glycyrrhizic acid FGE.36 SCF pinion adopted 2003 (Commission Directive 2004/77/EC (EC, 2004b)). JECFA evaluation 2004: Data were inadequate to derive an ADI. 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 pinion 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 FGE.79 Adopted EFSA consideration of JECFA evaluated substances. 17.018 Phenylalanine 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 lcysteine hydrochloride FGE.26Rev1 Adopted EFSA evaluation of amino acids. 17.033 lcysteine FGE.79 Adopted EFSA consideration of JECFA evaluated substances. 17.034 Glycine FGE.79 Adopted EFSA consideration of JECFA evaluated substances. f 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): FGE.29: Aromatic hydrocarbon: vinylbenzene [FLno: 01.015]. The EFSA Journal (2008) 740, 619

FGE.35: Quinoline alkaloids: quinine hydrochloride [FLno: 14.011], quinine sulphate [FLno: 14.152], quinine monohydrochloride dihydrate [FLno: 14.155]. 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]. ACKNWLEDGEMENT 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. TERMS F REFERENCE The 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). The EFSA Journal (2008) 740, 719

ASSESSMENT 1. Presentation of the Substances in the Flavouring Group Evaluation 36 1.1. Description The present Flavouring Group Evaluation 36 (FGE.36), using the procedure as referred to in the Commission Regulation (EC) No 1565/2000 (EC, 2000a) (The Procedure shown in schematic form in Annex I), deals with two triterpene glycosides, glycyrrhizic acid [FLno: 16.012] and glycyrrhizic acid, ammoniated [FLno: 16.060] from chemical group 30, Annex I of Commission Regulation (EC) No 1565/2000 (EC, 2000a). The two 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.). Glycyrrhizic acid [FLno: 16.012] and glycyrrhizic acid, ammoniated [FLno: 16.060] can exist as two stereoisomers (18alpha and 18betaisomer). The two substances have been presented without specification of the stereoisomeric composition. 1.3. Natural ccurrence in Food Glycyrrhizic acid is a natural constituent of liquorice, isolated from the dried root of the plant Glycyrrhiza glabra L. Liquorice root contains about 215 % glycyrrhizic acid and liquorice blocks and extract powder about 425 % (Størmer et al., 1993). Glycyrrhizic acid occurs mainly as the 18 betaisomer but up to approximately 10 % can occur as the 18alphaisomer (Amagaya et al., 1985). 2. Specifications Purity criteria for the two 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, 2000a), the information is adequate for the two candidate substances. Stereoisomeric composition is lacking for both glycyrrhizic acid [FLno: 16.012] and glycyrrhizic acid, ammoniated [FLno: 16.060] (see Section 1.2 and Table 1). The EFSA Journal (2008) 740, 819

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). ne 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. ne 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 rganization 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 (IFI, 1995). The intake approach does not consider the possible natural occurrence in food. 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 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) 740, 919

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 glycyrrhizic acid [FLno: 16.012] and glycyrrhizic acid, ammoniated [FLno: 16.060] 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) 16.012 Glycyrrhizic acid 240 210000 Class III 90 16.060 Glycyrrhizic acid, ammoniated 130 210000* Class III 90 * The mtamdi calculation is based on the maximum use levels as no information on the normal use levels has been provided (see Table 3.2). Table 3.2 shows the normal and maximum use levels for the candidate substances. Table 3.2 Normal and Maximum use levels (mg/kg) for candidate substances in FGE.36 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 16.012 375 375 16.060 40 91 5000 1512 45 61 200 25 20 300 50 550 100 50 60 200 200 150 79 10 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 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 The EFSA Journal (2008) 740, 1019

4. Evaluation/ Regulatory Status Glycyrrhizic acid [FLno: 16.012] and glycyrrhizic acid, ammoniated [FLno: 16.060] have previously been evaluated by the Scientific Committee on Food (SCF, 2003b): Previously, the Committee evaluated the toxicological information for glycyrrhizinic acid and concluded that the data were inadequate to derive an ADI (SCF, 1991). At that time, the Committee considered it prudent that regular ingestion should not exceed 100 mg/day, while it was explicitly mentioned that this was a provisional figure. Since then, new toxicological information, including data from human volunteer studies, has become available. Although these data provide a stronger basis for the upper limit for regular ingestion of glycyrrhizinic acid of 100 mg/day, the Committee still is of the opinion that an ADI for glycyrrhizinic acid and ammonium glycyrrhizinate cannot be derived, because the new human toxicity studies are too limited (small experimental groups, short duration). The Committee considers that this upper limit for regular ingestion of 100 mg/day provides a sufficient level of protection for the majority of the population. It is noted that this upper limit includes the intake of glycyrrhizinic acid via all products, liquorice confectionery as well as glycyrrhizinic acid or ammonium glycyrrhizinateflavoured products. At the same time, the Committee realises that within the human population there are subgroups for which this upper limit might not offer sufficient protection. Glycyrrhizic acid [FLno: 16.012] and glycyrrhizic acid, ammoniated [FLno: 16.060] have also been evaluated by the JECFA (JECFA, 2004a). Annex II provides more detail on the outcome of these evaluations. 5. Conclusions The Panel agrees with the evaluation by the SCF and emphasises the need for further information on use levels and on intakes that were requested by the SCF in its pinion from 2003 (see Annex II). The mtamdi of 210 mg/person/day exceeds by a factor of two the intake of 100 mg/person/day considered by the SCF to be a sufficient level of protection. Therefore, refined use levels and intake data are needed. Furthermore, the two substances have been presented without specification of the stereoisomeric composition. The EFSA Journal (2008) 740, 1119

TABLE 1: SPECIFICATIN SUMMARY F THE SUBSTANCES IN THE FLAVURING GRUP EVALUATIN 36 Table 1: Specification Summary of the Substances in the Flavouring Group Evaluation 36 FLno EU Register name Structural formula FEMA no CoE no CAS no 16.012 Glycyrrhizic acid 6) H H 2528 2221 1405863 Phys. form Mol. formula Mol. weight Solid C 42H 62 16 822.93 Solubility 1) Solubility in ethanol 2) Boiling point, C 3) Melting point, C ID test Assay minimum 200 IR 95 % Refrac. Index 4) Spec. gravity 5) n.a. n.a. Specification comments SE 7), SW 8). Decomposes at 200 C. Stereoisomeric composition to be specified.. CASrn is 1405869. H H H H H H H H H 18betaisomer shown 16.060 Glycyrrhizic acid, ammoniated 6) H, NH 4 + 2528 2221 53956040 Solid C 42H 65N 16 839.96 Soluble 209 IR 95 % n.a. n.a. SE 7). Stereoisomeric composition to be specified. H H H H H H H H H H 18betaisomer shown 1) Solubility in water, if not otherwise stated. 2) Solubility in 95% ethanol, if not otherwise stated. The EFSA Journal (2008) 740, 1219

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) Stereoisomeric composition not specified. 7) SE: Missing data on solubility in ethanol. 8) SW: Missing data on solubility. The EFSA Journal (2008) 740, 1319

ANNEX I: PRCEDURE FR THE SAFETY EVALUATIN 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 pinion of the Scientific Committee on Food expressed on 2 December 1999 (SCF, 1999), which is derived from the evaluation procedure developed by the Joint FA/WH 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. ne 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 NAEL exist on the flavourings or on structurally related substances (Step A5 and B4)? 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) 740, 1419

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. The EFSA Journal (2008) 740, 1519

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 No Step B3. 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 substance or are its metabolites endogenous? Yes Substance would not be expected to be of safety concern Yes Does a NAEL exist for the substance which provides an adequate margin of safety under conditions of intended use, or does a NAEL exist for structurally related substances which is high enough to accommodate any perceived difference in toxicity between the substance and the related substances? Step A5. No Yes No Does a NAEL exist for the substance which provides an adequate margin of safety under conditions of intended use, or does a NAEL 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) 740, 1619

ANNEX II: EVALUATIN/ REGULATIN STATUS Glycyrrhizic acid [FLno: 16.012] and glycyrrhizic acid, ammoniated [FLno: 16.060] SCF pinion adopted 2003 (SCF, 2003b) The Committee was asked to consider if the pinion of the Committee expressed in 1991 on glycyrrhizin is still valid in the light of additional information resulting from toxicological and clinical studies published since then on both glycyrrhizinic acid and its salts. The Committee is asked to take into account dietary exposure from all known sources, including contributions due to its natural occurrence in liquorice and through the ingestion of food products to which it is added as a flavouring substance. The Committee is also asked to evaluate ammonium glycyrrhizinate as a chemically defined flavouring substance for the possible acceptability of its inclusion in the Community Register. Conclusion by the SCF: Previously, the Committee evaluated the toxicological information for glycyrrhizinic acid and concluded that the data were inadequate to derive an ADI (SCF, 1991). At that time, the Committee considered it prudent that regular ingestion should not exceed 100 mg/day, while it was explicitly mentioned that this was a provisional figure. Since then, new toxicological information, including data from human volunteer studies, has become available. Although these data provide a stronger basis for the upper limit for regular ingestion of glycyrrhizinic acid of 100 mg/day, the Committee still is of the opinion that an ADI for glycyrrhizinic acid and ammonium glycyrrhizinate cannot be derived, because the new human toxicity studies are too limited (small experimental groups, short duration). The Committee considers that this upper limit for regular ingestion of 100 mg/day provides a sufficient level of protection for the majority of the population. It is noted that this upper limit includes the intake of glycyrrhizinic acid via all products, liquorice confectionery as well as glycyrrhizinic acid or ammonium glycyrrhizinateflavoured products. At the same time, the Committee realises that within the human population there are subgroups for which this upper limit might not offer sufficient protection. These subgroups comprise people with decreased 11betahydroxysteroid dehydrogenase2 activity (the target enzyme of glycyrrhizinic acid, for which genetic polymorphisms resulting in reduced basal activity have been described), people with prolonged gastrointestinal transit time, and people with hypertension or electrolyterelated or water homeostasisrelated medical conditions. A more extensive discussion on sensitive subgroups in the population can be found in Annex II (section on modeling Pharmacokineticpharmacodynamic model ). The Committee notes that for ammonium glycyrrhizinate as well as for glycyrrhizinic acid, used as chemically defined flavouring substances, the Upper Use Levels in foods indicate that the Maximised SurveyDerived Intake (MSDI) exposure estimates (130 and 240 microg/person/day, respectively) may underestimate the intake for individuals who select to consume certain foods, e.g. foods flavoured at the Upper Use Levels (see Annex I). To complete the evaluation of glycyrrhizinic acid and ammonium glycyrrhizinate as chemically defined flavouring substances, the following information needs to be provided: The EFSA Journal (2008) 740, 1719

for glycyrrhizinic acid: specifications as required by Commission Regulation (EC) No. 1565/2000. for ammonium glycyrrhizinate: adequate data on solubility, as required by Commission Regulation (EC) No. 1565/2000. data on occurrence of the 18alpha isomer in the commercial product. The Committee notes that glycyrrhizinic acid and ammonium glycyrrhizinate are evaluated here irrespective of their chirality. for both glycyrrhizinic acid and ammonium glycyrrhizinate: more refined usage data (e.g market share data), as it seems these substances are used in many food categories, but within a given food category probably only in very few products. JECFA evaluation (JECFA, 2005c) The JECFA Committee was asked to comment on the safety of glycyrrhizinic acid and its monoammonium salt as a natural constituent of liquorice (USA, licorice ) and in its use as a flavouring substance in various food products. Conclusion by the JECFA: The Committee concluded that the safety evaluation of glycyrrizinic acid should be based on the data from humans. It was observed that there is a sensitive subset of the population who appear to show signs of pseudohyperaldosteronism at lower exposures than those which produce effects in the general population, but the available data did not allow the Committee to adequately characterize this subgroup, and hence the data could not be used to assign an ADI. The available data suggest that an intake of 100 mg/day would be unlikely to cause adverse effects in the majority of adults. The Committee recognized that, in certain highly susceptible individuals, physiological effects could occur at intakes somewhat below this figure. The data indicate that consumers with a high intake of liquorice confectionery or herbal tea containing liquorice may have an intake of glycyrrhizinic acid of >100 mg/day. EU Regulation (Commission Directive 2004/77/EC) Commission Directive 2004/77/EC of 29 April 2004 amending Directive 94/54/EC as regards the labelling of certain foods containing glycyrrhizinic acid and its ammonium salt, requiring that confectionery or beverages containing 100 mg/kg or 10 mg/l or above should be labelled contains liquorice. Confectionery containing 4 g/kg or above should be labelled contains liquorice. People suffering from hypertension should avoid excessive consumption. Beverages containing 50 mg/l or above or of 300 mg/l or above in the case of beverages containing more than 1.2 % by volume of alcohol should be labelled contains liquorice. People suffering from hypertension should avoid excessive consumption (EC, 2004b). The EFSA Journal (2008) 740, 1819

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