SCIENTIFIC OPINION. EFSA Panel on Additives and Products or Substances used in Animal Feed (FEEDAP) 2,3

Transcription

1 SCIENTIFIC OPINION Scientific Opinion on the safety and efficacy of vitamin B 1 (thiamine mononitrate) as a feed additive for all animal species based on a dossier submitted by VITAC EEIG 1 EFSA Panel on Additives and Products or Substances used in Animal Feed (FEEDAP) 2,3 ABSTRACT European Food Safety Authority (EFSA), Parma, Italy Thiamine mononitrate administered via feed or water for drinking is safe for the target animals having a wide margin of safety. The use of thiamine mononitrate as an additive in animal nutrition is safe for consumers. Since no data on inhalation toxicity of thiamine mononitrate has been provided, inhalation of dust is considered as potentially hazardous. Similarly no data is available on skin and eye irritation. Thiamine mononitrate is regarded as a skin and eye irritant. On the basis of published literature, thiamine should be considered as a sensitiser. A risk for the environment resulting from the use of thiamine mononitrate in animal nutrition is not foreseen. Thiamine mononitrate is regarded as an effective source of vitamin B 1. European Food Safety Authority, 2011 KEY WORDS Nutritional additive, vitamins and provitamins, vitamin B 1, thiamine mononitrate, safety 1 On request from the European Commission, Question No EFSA-Q , adopted on 11 October Panel members: Gabriele Aquilina, Georges Bories, Andrew Chesson, Pier Sandro Cocconcelli, Joop de Knecht, Noël Dierick, Mikolaj Antoni Gralak, Jürgen Gropp, Ingrid Halle, Christer Hogstrand, Reinhard Kroker, Lubomir Leng, Anne- Katrine Lundebye Haldorsen, Secundino Lopez Puente, Alberto Mantovani, Giovanna Martelli, Miklós Mézes, Derek Renshaw, Maria Saarela, Kristen Sejrsen, Johannes Westendorf. Correspondence: FEEDAP@efsa.europa.eu 3 Acknowledgement: The Panel wishes to thank the members of the Working Group on Vitamins, including Annette Schuhmacher, for the preparatory work on this scientific opinion. Suggested citation: EFSA Panel on Additives and Products or Substances used in Animal Feed (FEEDAP); Scientific Opinion on the safety and efficacy of vitamin B 1 (thiamine mononitrate) as a feed additive for all animal species based on a dossier submitted by VITAC EEIG.. [14 pp.]. doi: /j.efsa Available online: European Food Safety Authority, 2011

2 SUMMARY Following a request from the European Commission, the Panel on Additives and Products or Substances used in Animal Feed (FEEDAP) was asked to deliver a scientific opinion on the safety and efficacy of vitamin B 1 in the form of thiamine mononitrate as an additive to feed and water for drinking for all animal species. Thiamine is present in the organism predominantly as thiamine pyrophosphate, followed by thiamine triphosphate and thiamine monophosphate. Vitamin B 1 mainly acts in α-ketoacid decarboxylation (e.g. pyruvate, α-ketoglutarate and branched-chain α-ketoacid acids) and in transketolation (e.g. among hexose and pentose phosphates). Main deficiency symptoms are characterised as disorders in neural function. Oral administration routes of thiamine mononitrate via feed or water for drinking are considered as bioequivalent. Thiamine mononitrate is safe for the target animals, having a wide margin of safety. The use of thiamine mononitrate as an additive in animal nutrition is safe for consumers. Since no data on inhalation toxicity of thiamine mononitrate has been provided, inhalation of dust is considered as potentially hazardous. Similarly no data is available on skin and eye irritation. Thiamine mononitrate is regarded as a skin and eye irritant. On the basis of published literature, thiamine should be considered as a sensitiser. Thiamine occurs widely in nature. Its use in animal nutrition is not expected to substantially increase the concentration in the environment. Therefore, a risk for the environment resulting from the use of thiamine mononitrate in animal nutrition is not foreseen. Thiamine mononitrate is regarded as an effective source of vitamin B 1. To guarantee the quality of thiamine mononitrate, the FEEDAP Panel recommends that the specifications detailed in the European Pharmacopeia should be applied. 2

3 TABLE OF CONTENTS Abstract... 1 Summary... 2 Table of contents... 3 Background... 4 Terms of reference... 4 Assessment Introduction Characterisation Manufacturing process Stability and homogeneity Shelf life of the additive Stability in premixtures, feed and water for drinking Homogeneity Physico-chemical incompatibilities in feed Conditions of use Evaluation of the analytical methods by the European Union Reference Laboratory (EURL) Safety Safety for the target species Safety for the consumer Absorption, distribution, metabolism and excretion of thiamine Assessment of consumer safety Safety for the user Effects on the respiratory system Effects on the eyes and skin Sensitisation potential Safety for the environment Efficacy Post-market monitoring Conclusions and recommendations Documentation provided to EFSA References Appendix

4 BACKGROUND Regulation (EC) No 1831/ establishes the rules governing the Community authorisation of additives for use in animal nutrition. In particular, Article 4(1) of that Regulation lays down that any person seeking authorisation for a feed additive or for a new use of a feed additive shall submit an application in accordance with Article 7; in addition, Article 10(2) of that Regulation specifies that for existing products within the meaning of Article 10(1), an application shall be submitted in accordance with Article 7, at the latest one year before the expiry date of the authorisation given pursuant to Directive 70/524/EEC for additives with a limited authorisation period, and within a maximum of seven years after the entry into force of this Regulation for additives authorised without a time limit or pursuant to Directive 82/471/EEC. The European Commission received a request from the VITAC EEIG Vitamins Authorisation Consortium 5 for (i) authorisation of a new use (i.e. use in water for drinking) and (ii) re-evaluation of authorisation of vitamin B 1 in the form of thiamine mononitrate, when used as a feed additive for all animal species (category: nutritional additive; functional group: vitamins, provitamins and chemically well-defined substances having similar effect) under the conditions mentioned in Table 1. According to Article 7(1) of Regulation (EC) No 1831/2003, the Commission forwarded the application to the European Food Safety Authority (EFSA) as an application under Article 4(1) (authorisation of a feed additive or new use of a feed additive) and under Article 10(2)/(7) (reevaluation of an authorised feed additive). EFSA received directly from the applicant the technical dossier in support of this application. 6 According to Article 8 of that Regulation, EFSA, after verifying the particulars and documents submitted by the applicant, shall undertake an assessment in order to determine whether the feed additive complies with the conditions laid down in Article 5. The particulars and documents in support of the application were considered valid by EFSA as of 20 April The Scientific Committee on Food expressed an opinion on the tolerable upper intake level of vitamin B 1 (SCF, 2001). The Panel on Food Additives and Nutrient Sources added to food issued a scientific opinion on benfotiamine, thiamine monophosphate chloride and thiamine pyrophosphate chloride, as sources of vitamin B 1 added for nutritional purposes to food supplements (EFSA, 2008) and an opinion on the inability to assess the safety of thiamine-enriched yeast added for nutritional purposes as a source of thiamine in food supplements and the bioavailability of thiamine from this source, based on the supporting dossier (EFSA, 2009a). The Panel on Dietetic Products, Nutrition and Allergies (NDA) issued an opinion on substantiation of several health claims pursuant to Article 13(1) of Regulation (EC) No 1924/2006 (EFSA, 2009b) and another opinion on the substantiation of a health claim related to thiamine and carbohydrate and energy-yielding metabolism pursuant to Article 14 of Regulation (EC) No 1924/2006 (EFSA, 2010). Vitamin B 1 in the form of thiamine mononitrate has been authorised without a time limit under Council Directive 70/524/EEC 8 for its use for all animal species as a nutritional additive. TERMS OF REFERENCE According to Article 8 of Regulation (EC) No 1831/2003, EFSA shall determine whether the feed additive complies with the conditions laid down in Article 5. EFSA shall deliver an opinion on the safety for the target animal(s), consumer, user and the environment and the efficacy of vitamin B 1 in the form of thiamine mononitrate, when used under the conditions described in Table OJ L 268, , p. 29. VITAC EEIG Vitamins Authorisation Consortium, 130A, B-1050 Brussel, Belgium. Company: Orffa International Holding BV, The Netherlands. EFSA Dossier reference: FAD A new mandate was received in EFSA in March OJ C 50, , p. 1. 4

5 Table 1: Description and conditions of use of the additive as proposed by the applicant Additive Registration number/ec No/No (if appropriate) Category(ies) of additive Functional group(s) of additive Vitamin B 1 / Thiamine mononitrate -- Nutritional additives Vitamins, provitamins and chemically well defined substances having a similar effect Description Composition, description Chemical Purity criteria Method of analysis formula (if appropriate) (if appropriate) Thiamine mononitrate C 12 H 17 N 4 OS.NO 3 Min. 98.0% Current European Pharmacopoeia method Trade name (if appropriate) -- Name of the holder of authorisation (if appropriate) -- Species or category of animal All animal species and categories Maximum Age Conditions of use Minimum content Maximum content mg or Units of activity or CFU/kg of complete feedingstuffs (select what applicable) Withdrawal period (if appropriate) Specific conditions or restrictions for use (if appropriate) Specific conditions or restrictions for handling (if appropriate) Post-market monitoring (if appropriate) Specific conditions for use in complementary feedingstuffs (if appropriate) Marker residue Other provisions and additional requirements for the labelling Only for manufacture of animal feed. Applicable in premix, feed & water. The additive may be incorporated as such or in the form of an additive preparation containing min. 96% thiamine mononitrate and approx. 2% anti-caking agent. -- No specific requirements other than the traceability and complaint system implemented in compliance with the requirements of Regulation No 183/2005. Applicable in premix, feed & water. Direct incorporation of the additives necessitates specific equipment in the production site, in order to ensure proper and homogeneous mixing. Maximum Residue Limit (MRL) (if appropriate) Species or category of animal Target tissue(s) or food products Maximum content in tissues

6 ASSESSMENT This opinion is based in part on data provided by a single company (representing two manufacturers) involved in the production of vitamin B 1 in the form of thiamine mononitrate and belonging to VITAC EEIG Authorisation Consortium. It should be recognised that this data covers a fraction of existing additives containing thiamine mononitrate. The composition of the additives is not subject of the application. The FEEDAP Panel has sought to use the data provided together with data from other sources to deliver an opinion and to produce recommendations for the authorisation, which would secure the safety of future uses of vitamin B 1 in the form of thiamine mononitrate as feed additive. 1. Introduction Thiamine is present in the organism predominantly as thiamine pyrophosphate, followed by thiamine triphosphate and thiamine monophosphate. Vitamin B 1 mainly acts in α-ketoacid decarboxylation (e.g. pyruvate, α-ketoglutarate and branched-chain α-ketoacid acids) and in transketolation (e.g. among hexose and pentose phosphates). Main deficiency symptoms are characterised as disorders in neural function. Vitamin B 1 in the form of thiamine mononitrate is included in the European Union Register of Feed Additives pursuant to Regulation (EC) No 1831/2003. It is authorised without a time limit in application of Article 9t (b) of Council Directive 70/524/EEC 9 concerning additives in feedingstuffs (2004/C 50/01) for it use in all animal species as a nutritional additive. The applicant, a consortium representing two manufacturers, asks for the re-evaluation of the use of vitamin B 1 in the form of thiamine mononitrate as additive to feed for all animal species and categories without restrictions on age, (withdrawal) time and content in feedingstuff. The applicant is also seeking for a new use of vitamin B 1 in the form of thiamine mononitrate in water for drinking. Vitamin B 1 as thiamine mononitrate is authorised for use in food (Regulation (EC) No 1925/2006, 10 amended by Regulation (EC) No 1170/2009) 11 and in food supplements (Directive 2002/46/EC, Annex II), 12 for addition for specific nutritional purposes in foods for particular nutritional uses (Regulation (EC) No 953/2009), 13 to processed cereal based foods and baby foods for infants and young children (Directive 2006/125/EC, Annex IV) 14 and to infant formulae and follow-on formulae when reconstituted as instructed by the manufacturer (Directive 2006/141/EC, Annex III). 15 Vitamin B 1 is also listed as a pharmacologically active substance in veterinary medicinal products and it is not subject to maximum residue levels when used in food-producing animals (Commission Regulation (EC) No 37/2010). 16 Thiamine mononitrate is authorised in cosmetics as skin conditioners (Commission Decision 2006/257/EEC). 17 Thiamine mononitrate is described in the European Pharmacopeia (PhEur) as Monograph (MG) Characterisation Thiamine mononitrate (IUPAC name 3-[(4-amino-2-methylpyrimidin-5-yl)methyl]-5-(2- hydroxyethyl)-4-methylthiazolium nitrate, synonyms: vitamin B 1, aneurine) is identified by the CAS number and the EINECS number The structural formula of thiamine mononitrate is shown in Figure 1. 9 OJ C 50, , p OJ L , p OJ L , p OJ L , p OJ L 269, , p OJ L , p OJ L , p OJ L 15, , p OJ L , p. 1. 6

7 Figure 1: Structural formula of thiamine mononitrate The molecular formula of thiamine mononitrate is C 12 H 17 N 4 OS NO 3 and its molecular weight is It has a melting point of 198 C (decomposition), shows a bulk density of approximately 0.35 g/ml and is soluble in water (2.7 g dissolves in about 100 ml at 25 C and approximately 30 g in 100 ml at 100 C), slightly soluble in methanol, ethanol and chloroform, insoluble in ether. Thiamine mononitrate is a white or almost white to yellowish crystalline powder, with a faint, characteristic odour and a bitter taste. PhEur MG 0531 requires a minimum of 98 % thiamine mononitrate in the dried substance. The analysis of five batches of each product showed an average (± SD) content of thiamine mononitrate of ± 0.04 % (product 1) 18 and ± 0.54 % (product 2). 19 Water was in the range %. Thiamine mononitrate of both manufacturers complies for both criteria with PhEur MG The amount of substance-related impurities as well as other impurities (sulphated ash and heavy metals) comply with the thresholds by PhEur, as demonstrated by the analysis of five batches of each producer. Data for residual solvents (methanol and ethanol) indicate that VICH thresholds are not exceeded. Control measures are in place. One batch from each manufacturer was analysed for particle size distribution determined by laser diffraction. The fraction of particles below 50 µm was between 70 % and 88 % (v/v). 20,21 The substance of one manufacturer was examined for its dusting potential which was 1.30 g/kg (corresponding to 3.2 g/m 3 ) Manufacturing process Thiamine mononitrate is produced by chemical synthesis out of aminopropionitrile, via a process involving several steps, i.e. formylation, condensation, cyclisation and oxidation. Intermediates of the chemical synthesis are alkene amide, formylpyrimidine and thiothiamine. The purification of thiamine mononitrate is a step-wise process including decolorisation and absorption on active carbon, microfiltration, crystallisation, centrifugation and washing. The applicant provided generic flow-charts of the synthetic process and the purification steps. A granular form of the additive is produced by the addition of an anticaking agent at the level of 2 % Stability and homogeneity Shelf life of the additive Thiamine mononitrate (nine batches from two producers) was demonstrated to have a shelf life of at least 36 (six batches) or 18 months (three batches) at 25 C and 60 % relative humidity (RH). Shelf life under accelerated conditions at 40 C (75 % RH) was shown for three batches of the product for 18 Technical dossier/section II/Annex Technical dossier/section II/Annex Technical dossier/section II/Annex Technical dossier/supplementary information March 2011/Annex B. 22 Technical dossier/supplementary information March 2011/Annex C. 7

8 six months. 23 The shelf life of the granular form (six batches) of thiamine mononitrate was demonstrated at 25 C and 60 % RH to be at least 36 months Stability in premixtures, feed and water for drinking According to the data published in the literature (Coelho, 2002), the stability of the additive in premixtures depends on the presence of trace elements and choline. Recovery of thiamine mononitrate after six months of storage in a vitamin premixture without choline chloride was 99 %, with choline chloride 73 %, in a vitamin trace element premixture without choline chloride 80 %, with choline chloride 48 %. These data indicate a reduced stability of thiamine mononitrate in the presence of choline chloride and of inorganic compounds of trace elements. The retention of thiamine mononitrate during feed processing is inversely correlated with the temperature applied during pelleting or extrusion. The recovery of thiamine mononitrate after pelleting was 96 % at C and 66 % at C, after extrusion 93 % at C and 66 % at C (Coelho, 2002). Similar results were obtained by Dozier (2002) for thiamine (form not given). The average monthly loss of thiamine mononitrate in pelleted and extruded feed can be estimated according to Coelho (1996) to be 4.5 %. The stability of thiamine mononitrate in water (one batch) was demonstrated when added at 10 mg/ml and kept for one week at room temperature Homogeneity Based on a statistical method (Jansen, 1992), in one example the coefficient of variation (CV %) for homogeneity of thiamine mononitrate in poultry feed was calculated to be around 1 %. Calculations for piglet, calf and cat feed, resulted in CVs of 0.16 %, 0.02 % and 0.12 %, respectively. However, this method has been developed to test the working accuracy of mixing equipments. Thiamine mononitrate is soluble in water at concentrations of 27 g/l at 25 C. Therefore, homogeneity in water for drinking does not need to be demonstrated Physico-chemical incompatibilities in feed No physico-chemical incompatibilities or interactions have been reported between thiamine mononitrate and feed materials, carriers, other approved additives or medicinal products when the additive was added to premixtures and feed. No such incompatibilities or interactions are expected Conditions of use Vitamin B 1 in the form of thiamine mononitrate is intended for use in all animal species and categories without a maximum content and a withdrawal period. The applicant states that the active substance can be administered via feed (premixtures, complete or complementary feed) and water for drinking Evaluation of the analytical methods by the European Union Reference Laboratory (EURL) EFSA has verified the EURL report as it relates to the methods used for the control of thiamine mononitrate in animal feed. The Executive Summary of the EURL report can be found in the Appendix. 23 Technical dossier/section II/Annexes 2.12 and Technical dossier/section II/Annex Technical dossier/section II/Annex

9 3. Safety Vitamin B 1 (thiamine mononitrate) for all animal species According to Regulation (EC) No 429/2008, tolerance, metabolism and residue, and toxicological (concerning consumer safety) studies are not required for vitamins, pro-vitamins and chemically defined substances having similar effects which are already authorised as feed additives under Directive 70/524/EEC and which do not have the potential to accumulate, which the FEEDAP Panel considers is the case for vitamin B Safety for the target species Little information is published concerning thiamine tolerance and toxicity in domestic animal species. Studies in mice, rats, rabbits and dogs indicate that parenteral administration ( mg/kg bw) of thiamine (hydrochloride) may be the only route by which signs of thiamine toxicity can be produced. In those animal species, thiamine toxicity is characterised by depression of the respiratory centre; however, the mechanism of those effects is unknown. The NRC (1987) concluded that dietary intake of thiamine up to 1000 times the requirement level are apparently safe for most animal species. Requirements for thiamine (NRC, see McDowell, 2000) are in the range of mg/kg feed for poultry, mg/kg for pigs, 1 15 mg/kg for fish and mg/kg for pets. Vitamin supplementation of commercial compound feed is mostly oriented towards recommendations, which are in the range of 1 4 mg/kg for pigs and poultry, 8 15 mg/kg for fish and 2 10 mg/kg feed for pets (AWT, 2002). A survey on vitamin supplementation of commercial feeds for pigs and poultry in Europe (Belgium, Denmark, Germany, Italy, Netherlands, Portugal, Spain, United Kingdom) identified a range of 0 7 mg vitamin B 1 /kg as commercial use levels (Gropp, 1994). A range for piglets and pigs of 1 3 mg/kg feed was reported by Whittemore et al. (2002). Three classes of thiamine antagonists are known. Structurally-related compounds (e.g. pyrithiamine, oxythiamine and amprolium) exert an antagonistic effect by a competitive mechanism. Structurally unrelated compounds are heat-stable polyphenols in feedingstuffs which oxidise the thiazole ring to non absorbable thiamine disulphide or sulphur which can reduce thiamine synthesis by bacteria. Thiaminases can be present in fresh fish or be produced by rumen microbes. Feed manufacturers are familiar with those thiamine antagonists and consider this in feed formulation. The FEEDAP Panel concludes that thiamine mononitrate is safe for the target animals having a wide margin of safety (about 1000 compared to the requirements/recommendations) Safety for the consumer Absorption, distribution, metabolism and excretion of thiamine Free thiamine released in the gastrointestinal tract is absorbed in the proximal intestinal tract by active and passive mechanisms. It is stored predominantly in the liver and kidneys. In humans 80 % of the body depot is found in form of thiamine pyrophosphate, 10 % as thiamine triphosphate and the remaining as thiamine monophosphate. Thiamine is excreted via the kidneys mainly in unchanged form, although several and partially different metabolites were identified in humans and rats. Thiamine is one of the most poorly stored vitamins. Mammals may deplete their body stores within one to two weeks (Ensminger et al., 1983). The only data found on the relationship between dietary thiamine and thiamine deposition in edible tissues and products refer to studies with laying hens. In one study (Leeson and Caston, 2003), the vitamin B 1 content was 49 and 67 µg/60 g egg after a three-month feeding period with diets supplemented with 4 and 24 mg thiamine/kg feed, respectively. Johnson and Korver (2008) found at lower dietary supplementation (2.4 and 11.4 mg thiamine/kg) after six weeks of feeding a higher thiamine concentration in the egg (239 and 395 µg/60 g egg). In another study (two factorial design, 60-week duration), thiamine in the whole egg was 0.52 and 0.65 mg/kg (corresponding to 31 and 39 µg/60 g egg) in groups fed diets supplemented with 0.91 and 3.0 mg/kg, respectively (Pérez-Vendrell et al., 2004). 9

10 In all studies, no data were provided on the background content of thiamine. Cereals may contain between 1 and 5 mg/kg and yeasts between 40 and 50 mg/kg; however literature data indicate considerable variation. Although all studies demonstrated the dose-dependent effect on thiamine content of eggs and considering the variable concentration in feed, the differences in the absolute values between the studies are so high that a considerable influence of the dietary thiamine level on the egg content of thiamine should not be expected in practice. It is concluded that vitamin B 1 supplementation of feed at recommended levels (see Section 3.1) would not essentially modify the contribution of thiamine from food of animal origin to consumer intake Assessment of consumer safety Estimates based on food intake indicate that reported mean intake of vitamin B 1 in some European countries varied from 1.10 mg/day to 2.28 mg/day with the high intake level (97.5 th percentile) ranging from 1.90 to 6.35 mg/day (SCF, 2001). The Scientific Committee on Food (SCF, 2001) could not identify a tolerable upper intake level (UL) of vitamin B 1 since no NOAEL could be established due to the lack of systematic oral dose-response intake studies as well as the extreme low toxicity. The Expert Group on Vitamins and Minerals (EVM, 2003) concluded that dietary supplements up to 100 mg/person/day (equivalent to 1.7 mg/kg bw/day supplemented thiamine for a 60-kg adult) would not be expected to result in adverse effects. This conclusion was based on the results of a randomised double-blind placebo-controlled study by Gokhale (1996) on 556 young females (12 21 years). No adverse effects were found when a daily oral dose of 100 mg of thiamine hydrochloride was given for 90 days followed by placebo for 60 days or vice versa. An exposure assessment is consequently not necessary since a contribution of thiamine in feed to the consumer intake is marginal and available data showed that the oral toxicity is very low and there is no UL for vitamin B 1 identified. Therefore, the FEEDAP Panel considers that the use of thiamine mononitrate and thiamine hydrochloride as additives in animal nutrition is safe for consumers Safety for the user Effects on the respiratory system Since no data on inhalation toxicity of thiamine mononitrate has been provided, inhalation of dust is considered as potentially hazardous Effects on the eyes and skin Because of the lack of data, thiamine mononitrate is considered as a skin and eye irritant Sensitisation potential Allergic eczematous reactions to topical exposure to thiamine hydrochloride in pharmaceutical workers have been reported (Larsen et al., 1989). Contact dermatitis may occur with occupational exposure and may cause sensitisation and lead to dermatitis-type reactions after oral or injected administrations. Anaphylactic reactions have only been observed with large parenteral (intravenous, intramuscular, subcutaneous) doses of thiamine hydrochloride, generally after multiple doses (Natural Standard Research Collaboration, 2010). Occupational asthmatic reactions have been reported (Seed and Agius, 2006). Thiamine (vitamin B 1 ) is the vitamin that most frequently induces an allergic reaction (Schalch and Dostalova, 1983 according to Fernandez et al. 1997). On this basis, thiamine should be considered as a sensitiser. 10

11 3.4. Safety for the environment Vitamin B 1 (thiamine mononitrate) for all animal species Thiamine occurs widely in nature. Its use in animal nutrition is not expected to substantially increase the concentration in the environment. Therefore, a risk for the environment resulting from the use of thiamine mononitrate in animal nutrition is not foreseen. 4. Efficacy According to Regulation (EC) No 429/2008, efficacy studies are not required for vitamins, provitamins and chemically defined substances having similar effects which are already authorised as feed additives under Directive 70/524/EEC and which do not have the potential to accumulate, which the FEEDAP Panel considers is the case for vitamin B 1. Due to the long history of use and its established nutritional role in domestic animals, thiamine mononitrate is regarded as an effective source of vitamin B 1. Vitamin B 1 has been globally used in animal nutrition for decades. Data on requirement, allowances and recommendations for feed supplementation are easily accessible as standard literature for animal nutrition experts. 5. Post-market monitoring The FEEDAP Panel considers that there is no need for specific requirements for a post-market monitoring plan other than those established in the Feed Hygiene Regulation 26 and Good Manufacturing Practice. CONCLUSIONS AND RECOMMENDATIONS CONCLUSIONS Oral administration routes of thiamine mononitrate via feed or water for drinking are considered as bioequivalent. Thiamine mononitrate is safe for the target animals, having a wide margin of safety. The use of thiamine mononitrate as an additive in animal nutrition is safe for consumers. Since no data on inhalation toxicity of thiamine mononitrate has been provided, inhalation of dust is considered as potentially hazardous. Similarly no data is available on skin and eye irritation. Thiamine mononitrate is regarded as a skin and eye irritant. On the basis of published literature, thiamine should be considered as a sensitiser. Thiamine occurs widely in nature. Its use in animal nutrition is not expected to substantially increase the concentration in the environment. Therefore, a risk for the environment resulting from the use of thiamine mononitrate in animal nutrition is not foreseen. Due to the long history of use and the established nutritional role in domestic animals, thiamine mononitrate is regarded as an effective source of vitamin B 1. RECOMMENDATIONS The FEEDAP Panel proposes to use the specifications for thiamine mononitrate according to PhEur (MG 0531) considering purity, substance-related impurities and other impurities (sulphated ash and heavy metals). When giving a warranty for stability in premixtures, the manufacturer should consider the effect of choline chloride. The FEEDAP Panel recommends including a corresponding warning under other 26 OJ L 35, , p

12 provisions of the authorisation (e.g. Stability of thiamine mononitrate may be reduced in premixtures containing choline chloride ). Considering the potential risk of users by inhalation exposure and to guarantee homogeneous distribution in the final feed, the use of thiamine mononitrate should be restricted to premixture manufacturers. Poor distribution of vitamin B 1 in feed may result in thiamine deficiency which would affect animal welfare. DOCUMENTATION PROVIDED TO EFSA 1. Vitamin B 1 (thiamine mononitrate) as a feed additive for all animal species. July Submitted by Submitted by VITAC EEIG Vitamins Authorisation Consortium. 2. Vitamin B 1 (thiamine mononitrate) as a feed additive for all animal species. Supplementary information. March Submitted by Submitted by VITAC EEIG Vitamins Authorisation Consortium. 3. Vitamin B 1 (thiamine mononitrate) as a feed additive for all animal species. Supplementary information. September Submitted by Submitted by VITAC EEIG Vitamins Authorisation Consortium. 4. Evaluation report of the European Union Reference Laboratory for Feed Additives on the methods(s) of analysis for thiamine monitrate. 5. Comments from Member States received through the ScienceNet. REFERENCES AWT (Arbeitsgemeinschaft für Wirkstoffe in der Tierernährung e.v.), Vitamins in animal nutrition, ISBN Coelho M, Stability of vitamins affected by feed processing. Feedstuffs, 29, Coelho M, Vitamin stability in Premixes and Feeds. A Practical Approach in Ruminant Diets. Proceedings 13 th Annual Florida Ruminant Nutrition Symposium, pp Dozier WA, Mash conditioning and vitamin stability. Feed International, May 2002, EFSA (European Food Safety Authority), Opinion of the Scientific Panel on Food Additives and Nutrient Sources added to food on Benfotiamine, thiamine monophosphate chloride and thiamine pyrophosphate chloride, as sources of vitamin B 1 added for nutritional purposes to food supplements. The EFSA Journal, 864, EFSA (European Food Safety Authority), 2009a. Opinion of the Scientific Panel on Food Additive and Nutrient Sources added to food on the inability to assess the safety of thiamine-enriched yeast added for nutritional purposes as a source of thiamine in food supplements and the bioavailability of thiamine from this source, based on the supporting dossier. The EFSA Journal, 1121, 1-6. EFSA Panel on Dietetic Products, Nutrition and Allergies (NDA), 2009b. Scientific opinion of the on substantiation of health claims related to thiamine and energy-yielding metabolism (ID 21, 24, 28), cardiac function (ID 20), function of the nervous system (ID 22, 27), maintenance of bone (ID 25), maintenance of teeth (ID 25), maintenance of hair (ID 25), maintenance of nails (ID 25), maintenance of skin (ID 25) pursuant to Article 13(1) of Regulation (EC) No 1924/2006. EFSA Journal, 7(9): EFSA Panel on Dietetic Products, Nutrition and Allergies (NDA), Scientific opinion on the substantiation of a health claim related to thiamine and carbohydrate and energy-yielding metabolism pursuant to Article 14 of Regulation (EC) No 1924/2006. EFSA Journal, 8(7):

13 Ensminger AH, Ensminger ME, Konlande JE and Robson JRK, In: Foods and Nutrition Encyclopedia, Vol. 1, A-H. California: Pegus Press, p European Pharmacopeia (PhEur), Thiamine mononitrate, Monograph (MG) th Edition. Strasbourg, France. Council of Europe (COE) European Directorate for the Quality of Medicines. EVM (Expert group on vitamins and minerals), 2003, online. Safe upper levels for vitamins and minerals. Available from Fernandez M, Barcelò M, Munoz C, Torrecillas M and Blanca M, Anaphylaxis to thiamine (vitamin B 1 ). Allergy, 52, Gokhale LB, Curative treatment of primary (spasmodic) dysmenorrhoea. Indian Journal of Medical Research, 103, Gropp JM Vitamin Fortification Levels in European Commercial Poultry and Swine Diets. Proceedings of the Arkansas Nutrition Conference, Fayetteville, Arkansas, Jansen HD, Mischtechnik im Futtermittelbetrieb. Anforderungen an Mischenlage, Arbeits- und Mischgenauigkeit. Die Mühle+ Mischfuttertechnik, 129, Johnson ML and Korver DR, 2008, online. The effect of dietary supplementation on egg vitamin content. Metabolism and Nutrition IV: Vitamins and Minerals and Feed Manufacturing, Abstracts of papers, Poultry Science Association, Niagara Falls, Ontario, Canada. Available from Larsen AI, Jepsen JR and Thulin H, Allergic contact dermatitis from thiamine. Contact Dermatitis, 20, Leeson S and Caston LJ, Vitamin Enrichment of Eggs. Journal of Applied Poultry Research, 12, McDowell LR Vitamins in animal and human nutrition, Second Edition, Iowa State University Press. ISBN , p NRC (National Research Council), Vitamin tolerance of animals. National Academic Press, Washington DC, Natural Standard Research Collaboration, 2010, online. Natural Standard Patient Monograph. Thiamin (thiamine), vitamin B 1. Available from Pérez-Vendrell M, L. Llauradó L, Brufau J, Hernández JM, Improving the nutritive value of eggs by feeding laying hens an Optimum Vitamin Nutrition (OVN ). Zootecnica, December, SCF (Scientific Committee on Food), 2001, online. Opinion of the Scientific Committee on Food on the Upper Tolerable Intake of Vitamin B 1. Available from Seed MJ and Agius RM, Prediction of asthma hazard of thiamine. Allergy, 61, 648. Schalch W and Dostalova L, Vitamins. In: de Week AL, Bundgard H, editors. Allergic reactions to drugs. Vol. 63. Berlin: Springer-Verlag, Whittemore CT, Close WH, Hazzledine MJ, The need for nutrient requirement standards for pigs. A report of the British Society of Animal Science nutritional standards working group: pigs. Pig News and Information, 23, 67N-74N. 13

14 APPENDIX Executive Summary of the Evaluation Report of the European Union Reference Laboratory for Feed Additives on the Method(s) of Analysis for thiamine mononitrate 27 In the current application authorisation is sought for Vitamin B 1 under the category/functional group 3(a) nutritional additives / vitamins, pro-vitamins and chemically well defined substances having similar effect according to the classification system of Regulation (EC) No 1831/2003. Specifically, authorisation is sought to use the active substance, thiamine mononitrate (Vitamin B 1 ) for all animal species and categories. The feed additive is a crystalline or granular powder with a minimum content of 96% of Vitamin B 1. It is intended to be incorporated directly in feedingstuffs (including complete feedingstuffs or complementary feedingstuffs) or through premixtures. Due to enough solubility in water the additive can be used directly in water. However, the Applicant does not propose any maximum or minimum concentration of feed additive in feedingstuffs or water. For the determination of the Vitamin B 1 in the feed additive the Applicant proposed the method described in the European Pharmacopoeia, based on High-Performance Liquid Chromatography (HPLC). The CRL considers this method suitable to be used within the frame of official control. For the determination of Vitamin B 1 in premixtures and feedingstuffs, the Applicant submitted a method described in the literature, based on Reverse Phase High-Performance Liquid Chromatography (RP-HPLC) coupled to a fluorescence detector, using an excitation wavelength at 360 nm and an emission wavelength at 430 nm. This method was further ring-trial validated and published in the Italian Official Journal. The following performance characteristics were reported for premixtures and feedingstuffs samples with a Vitamin B 1 content ranging from 7 to 484 mg/kg: - a relative standard deviation of repeatability (RSD r ) ranging from of 4.2 to 4.7%; - a relative standard deviation of reproducibility (RSD R ) ranging from 5 to 13%; - a recovery rate (R Rec ) ranging from 88 to 97%, and - a limit of quantification (LOQ) of 5 mg/kg. Based on these performance characteristics presented the EURL recommends the ring-trail validated method based on Reverse Phase High-Performance Liquid Chromatography (RP-HPLC) coupled to fluorescence detector for official control to determine Vitamin B 1 in premixtures and feedingstuffs. For the determination of Vitamin B 1 in the water, the Applicant proposed the CEN method EN 14122:2003, intended for foodstuffs. Due to enough solubility of the product in water, this method seems to be adequate to analyze thiamine mononitrate in water. However no experimental data were provided by the applicant for the aqueous media. Therefore the EURL cannot evaluate nor recommend the method for official control to determine Vitamin B 1 in water. Further testing or validation of the methods to be performed through the consortium of National Reference Laboratories as specified by article 10 (Commission Regulation (EC) No 378/2005) is not considered necessary. 27 The full report is available on the EURL website pdf 14