Abstract
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 l‐histidine monohydrochloride (HCl) monohydrate produced by fermentation using Corynebacterium glutamicum KCCM 80179 when used as a nutritional additive (amino acid) and as a sensory additive (flavouring compound) in feed and water for drinking for all animal species. The production strain is not genetically modified. No viable cells of the production strain were detected in the final product. The use of l‐histidine monohydrochloride monohydrate produced by fermentation using C. glutamicum KCCM 80179 is safe for the target species when used as a nutritional additive to supplement the diet in appropriate amounts to cover the requirements, depending on the species, the physiological state of the animal, the performance level, the environmental conditions, the background amino acid composition of the unsupplemented diet and the status of some essential trace elements such as copper and zinc. This conclusion would also cover the use as a sensory additive. l‐Histidine HCl monohydrate produced using C. glutamicum KCCM 80179 supplemented at levels appropriate for the requirements of species and life stage is considered safe for the consumer. l‐Histidine HCl monohydrate produced using C. glutamicum KCCM 80179 is not irritant to skin, is mildly irritant to eyes, and it is not a skin sensitiser. The additive does not pose a risk to users by inhalation. The use of l‐histidine HCl monohydrate produced by C. glutamicum KCCM 80179 in animal nutrition is not expected to represent a risk to the environment. l‐Histidine HCl monohydrate is considered an efficacious source of the essential amino acid l‐histidine for non‐ruminant animal species. For the supplemental l‐histidine to be as efficacious in ruminants as in non‐ruminant species, it would require protection against degradation in the rumen. It is also considered efficacious as a feed flavouring compound under the proposed conditions of use.
Keywords: nutritional additive, amino acid, l‐histidine monohydrochloride monohydrate, Corynebacterium glutamicum KCCM 80179, feed additive, safety
1. Introduction
1.1. Background and Terms of Reference
Regulation (EC) No 1831/20031 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.
The European Commission received a request from CJ Europe GmbH2 for authorisation of the product l‐histidine monohydrochloride monohydrate (feed grade) when used as a feed additive for all animal species (category: nutritional additives; functional group: amino acids, their salts and analogues; and category: sensory additives; functional group: flavourings).
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). The particulars and documents in support of the application were considered valid by EFSA as of 18 September 2018.
According to Article 8 of Regulation (EC) No 1831/2003, 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. EFSA shall deliver an opinion on the safety for the target animals, consumer, user and the environment and on the efficacy of the product L‐histidine monohydrochloride monohydrate produced by fermentation with Corynebacterium glutamicum KCCM 80179, when used under the proposed conditions of use (see Section 3.1.4).
1.2. Additional information
l‐Histidine monohydrochloride (HCl) monohydrate produced by fermentation with a non‐genetically modified strain of C. glutamicum (KCCM 80179) has not been assessed as a feed nutritional additive. The active substance of the product under application is l‐histidine.
l‐Histidine HCl monohydrate (minimum 98% on dry matter basis) produced by Escherichia coli ATCC 9637 is currently listed in the European Union Register of Feed Additives, and thus authorised in the European Union for use in feed for salmonids.3 l‐Histidine [EU Flavour Information System (FLAVIS) numbers 17.008] produced by chemical synthesis is currently listed in the European Union Register of Feed Additives, and thus authorised in the European Union as a feed flavouring.4
The EFSA Panel on Additives and Products or Substances used in Animal Feed (FEEDAP) assessed l‐histidine HCl monohydrate produced by E. coli ATCC 21318 as a nutritional feed additive (amino acid) for salmonids (EFSA, 2005, 2006a). The FEEDAP Panel assessed the safety and efficacy of l‐histidine as feed flavouring (EFSA FEEDAP Panel, 2014). The EFSA's Scientific Panel on Food Additives, Flavourings, Processing Aids and Materials in Contact with Food (AFC) evaluated l‐histidine and considered it safe for use as flavours in food (EFSA, 2006b, 2008a,b; EFSA CEF Panel, 2011).
l‐Histidine is authorised for use in food,5 cosmetics6 and as a veterinary medicinal product.7 , 8
l‐Histidine HCl monohydrate is described in a monograph of the European Pharmacopoeia (2017), monograph 01/2017:0910.
2. Data and methodologies
2.1. Data
The present assessment is based on data submitted by the applicant in the form of a technical dossier9 in support of the authorisation request for the use of l‐histidine HCl monohydrate produced by fermentation using C. glutamicum KCCM 80179 as a feed additive.
The FEEDAP Panel used the data provided by the applicant together with data from other sources, such as previous risk assessments by EFSA or other expert bodies, peer‐reviewed scientific papers, other scientific reports and experts’ knowledge, to deliver the present output.
EFSA has verified the European Union Reference Laboratory (EURL) report as it relates to the methods used for the control of the l‐histidine monohydrochloride monohydrate produced by fermentation with C. glutamicum KCCM 80179 in animal feed. The Executive Summary of the EURL report can be found in Annex A.10
2.2. Methodologies
The approach followed by the FEEDAP Panel to assess the safety and the efficacy of L‐histidine monohydrochloride monohydrate produced by fermentation with C. glutamicum KCCM 80179 is in line with the principles laid down in Regulation (EC) No 429/200811 and the relevant guidance documents: Guidance on the identity, characterisation and conditions of use of feed additives (EFSA FEEDAP Panel, 2017a), Guidance on the assessment of the safety of feed additives for the target species (EFSA FEEDAP Panel, 2017b), Guidance on the assessment of the safety of feed additives for the consumer (EFSA FEEDAP Panel, 2017c), Guidance on studies concerning the safety of use of the additive for users/workers (EFSA FEEDAP Panel, 2012), Guidance on the characterisation of microorganisms used as feed additives or as production organisms (EFSA FEEDAP Panel, 2018a), Guidance on the safety of feed additives for the environment (EFSA, 2008c), and Guidance on the assessment of the efficacy of feed additives (EFSA FEEDAP Panel, 2018b).
3. Assessment
The current application is for the authorisation of l‐histidine monohydrochloride monohydrate (minimum 98% purity) produced by fermentation using a ■■■■■ strain of C. glutamicum (KCCM 80179). The product is intended to be used in feed and water for drinking for all animal species as a nutritional additive (functional group: amino acids, their salts and analogues) and as a sensory additive (functional group: flavourings compounds).
3.1. Characterisation
3.1.1. Characterisation of the production organism
The production organism C. glutamicum has been deposited ■■■■■■■■■■ It is a ■■■■■■■■■■
The identity of the production strain as C. glutamicum was demonstrated ■■■■■■■■■■■■■■■.
The susceptibility of the production strain to the antibiotics listed in ■■■■■■■■■■ The minimum inhibitory concentrations were below to the cut off values set in such guidance.
3.1.2. Manufacturing process
l‐Histidine is produced by fermentation of the production strain. ■■■■■■■■■■
■■■■■■■■■■
3.1.3. Characterisation of the product/active substance
l‐Histidine monohydrochloride monohydrate (International Union of Pure and Applied Chemistry (IUPAC) name (2S)‐2‐amino‐3‐(1H‐imidazol‐5‐yl)propanoic acid;hydrate;hydrochloride, and synonyms l‐α‐Amino‐β‐(4‐imidazolyl)propionic acid monohydrochloride, glyoxaline‐5‐alanine hydrochloride) has the Chemical Abstracts Service (CAS) No 5934‐29‐2 and European Inventory of Existing Commercial Chemical Substances (EINECS) No 211‐438‐9. The chemical formula is C3H3N2–CH2–CH(NH2)–COOH· HCl·H2O and the molecular weight 209.63 g/mol. The structural formula is given in Figure 1.
Figure 1.
Molecular structure of l‐histidine monohydrochloride monohydrate
According to the specification, the additive contains ≥ 98% l‐histidine monohydrochloride monohydrate from which ≥ 73% is l‐histidine, ≤ 10% moisture, and ≤ 1% ash.18
Analysis of five batches showed an average histidine content of 73.5% ‘as is’ (range 73.5–73.6%), a chloride content of 16.7% (range 16.6–16.9%), a moisture content of 9.1% (range 8.9–9.3%), an ash content of 0.04% (range 0.02–0.07%) and 0.01% of phosphate (in all five batches).19 Free amino acids, ammonium, nitrate, nitrite, betaine and organic acids were not detected. The sum of the identified material on ‘as is’ basis was 99.4% (range 99.3–99.6%)
The specific optical rotation of the additive was measured in five batches of the final product and the average was + 10.48˚ (range + 10.42 to + 10.52˚).20 The measured values were within the range of the European Pharmacopoeia (+ 9.2 to + 10.6˚) and confirmed the l‐stereoisomer of histidine.
3.1.3.1. Impurities
Three batches of the additive were analysed for undesirable substances.
Heavy metals, (lead, cadmium and mercury) and arsenic were below the limits of detection (LODs).21 In the same batches, polychlorinated dibenzodioxins (PCDDs), and polychlorinated dibenzofurans (PCDFs) and coplanar dioxin‐like polychlorinated biphenyls (Co‐planar PCBs) were found below the LOD.22 Aflatoxins (B1, B2, G1, G2), ochratoxin A, zearalenone, deoxynivalenol, fumonisins B1 and B2 were below the corresponding LOD.23
In the same batches, nitrofurans (furazolidone, furaltadone, nitrofurazone and nitrofurantoin) and nitrofuran metabolites were below the corresponding LODs.24 A multiresidue pesticide analysis showed that none of the 358 pesticides was present in the three batches.25
Analysis of microbial contamination of the final product (three batches) indicated that Salmonella spp. (25 g sample), E. coli, and coliforms were absent whereas total bacterial count, was < 103 colony forming unit (CFU)/g; and yeasts and filamentous fungi were < 50 CFU/g.20
The presence of viable cells of the production strain in the final product was tested in three batches of the additive.■■■■■■■■■■ no bacterial growth was observed.
3.1.3.2. Physical properties
l‐Histidine monohydrochloride monohydrate is an off‐white, odourless, colourless and crystalline powder, with a bulk density of 550–750 kg/m3, melting point at 254°C, pH 3.5–4.5 (1% solution in water) and a water solubility of 56.6 g/L at 25°C.27
No information on the dusting potential of the additive under assessment was submitted. The particle size distribution (three batches analysed by sieving) showed that the fraction of particles < 125 μm ranged from 15 to 20% (w/w).28
3.1.3.3. Stability and homogeneity
The applicant submitted data on stability of an l‐histidine HCl monohydrate produced by a different strain of C. glutamicum (KCCM 80172). These data related to shelf life studies, stability in premixtures and feedingstuffs, in water for drinking and studies on the capacity of the additive to distribute homogeneously in feed (EFSA FEEDAP Panel, 2019).29 As the characteristics of the additives were similar and the production process was the same, the FEEDAP Panel considered the results of such studies applicable to the stability and the capacity to distribute homogeneously in feed of the additive under assessment. Regarding the shelf life, no losses were observed after storage at 25 or 40°C for 6 months. In a premixture with choline chloride, the losses ranged 4–6% after 6 months storage. In a mash feed, losses from 0 to 2% were observed after 3 months. No losses were observed in water for drinking stored at 25 and 40°C for 48 h. The coefficient of variation in a premixture was 5% and in a pelleted feed for chickens for fattening 6%.
The applicant provided a new stability study in feedingstuffs. The stability of three batches of the additive was studied in a pelleted feed for chicken for fattening (feed ingredients not described, only the nutritional composition) when supplemented at 0.4%.30 The additive was added via a premixture that was prepared containing 16% l‐histidine HCl monohydrate. The feed was pelleted at 72°C and dried at 60–65°C for 10 min. Feed processing (pelleting) represented a loss ranging from 3% to 8%. Pelleted samples were then kept at 25 ± 2°C packed in polyethylene bags for 3 months. At the end of the stability period, only one batch showed a loss of 16%.
3.1.3.4. Physicochemical incompatibilities in feed
No physicochemical incompatibilities in feed are expected with other additives, medicinal products or other feed materials.
3.1.4. Conditions of use
According to the applicant, the additive can be added directly in compound feed, through complementary feed or through premixtures and is aimed for all animal species. No proposed inclusion levels are provided, as the optimal daily allowance in quantitative terms depends on the species, the physiological state of the animal, the performance level and the environmental conditions, in particular on the amino acid composition of the unsupplemented diet.31
The additive can be also used in water for drinking.32
l‐Histidine monochloride monohydrate is proposed to be as a feed flavouring in feed or in water for drinking at a typical inclusion rate of 5 mg/kg, being the maximum recommended level of inclusion 25 mg/kg.
3.2. Safety
3.2.1. Safety for the target species
The absorption, distribution, metabolism and excretion of l‐histidine; its essentiality in animal nutrition; the requirements for the main food producing animal species, the histidine content in different feedingstuffs, and the consequences of excesses of histidine supplementation in animal nutrition were considered in a previous opinion of the EFSA FEEDAP Panel (2019).
Tolerance studies are not normally required for highly purified amino acids. Such tolerance studies with a certain indispensable amino acid will inevitably result in amino acid imbalances with depression of feed intake and hence impaired performance. No tolerance studies were submitted, and no use levels were proposed when used as a nutritional additive by the applicant. The applicant based the safety of the additive on the fact that l‐histidine HCl monohydrate is produced by a C. glutamicum that qualifies for the qualified presumption of safety (QPS) assessment, and has a high purity, with > 99% identified material.
On request of EFSA, the applicant provided a literature search for studies to address the safety for target animals. Several platform databases (Livivo, Toxnet, Ovid, Web of Knowledge, Google Scholar) were searched using the following search syntax: Histidine AND diet* AND (Chick* OR Birds OR Turkey OR Calf OR Calv* OR cow* OR rumen OR pig* OR fish OR salmon). The full text search for histidine in ‘title + keywords’ gave in total 658 hits. This was narrowed down to the term histidine in publication title: 89 hits. The search syntax “trace mineral” OR copper OR Zinc provided 23 hits. The same dataset assessed in a previous opinion (EFSA FEEDAP Panel, 2019) was submitted. Nearly all the studies found referred to requirements for l‐histidine and not to excesses nor tolerance for l‐histidine.
Although there is limited evidence from the published literature on the effects of supplementing histidine levels above the requirements, the FEEDAP Panel considers that, as with other amino acids, adverse effects might occur with levels of histidine in feeds exceeding the requirements, depending on the balance with other amino acids and the status of some essential trace elements such as copper and zinc.
The FEEDAP Panel, in its previous statement (EFSA FEEDAP Panel, 2010), identified risks of nutritional imbalances and hygienic concerns for amino acids when administered in water for drinking.
Since the levels proposed for the use of l‐histidine monohydrochloride monohydrate as flavouring (up to 25 mg/kg complete feed) are substantially lower than the animal requirements, the FEEDAP Panel considers l‐histidine monohydrochloride monohydrate produced with C. glutamicum KCCM 80179 is safe when used as a flavouring compound.
3.2.1.1. Conclusions on the safety for the target species
The use of l‐histidine monohydrochloride monohydrate produced by fermentation using Corynebacterium glutamicum KCCM 80179 is safe for the target species when used as a nutritional additive to supplement the diet in appropriate amounts to cover the requirements, depending on the species, the physiological state of the animal, the performance level, the environmental conditions, the background amino acid composition of the unsupplemented diet and the status of some essential trace elements such as copper and zinc. This conclusion would also cover the use as sensory additive.
3.2.2. Safety for the consumer
The product under assessment is produced by fermentation using a strain C. glutamicum (KCCM 80179) which fulfils the QPS qualifications for production purposes (EFSA BIOHAZ Panel, 2019). Therefore, the FEEDAP Panel considers that no safety concerns would derive from the fermentation process. The additive contains 98.9% l‐histidine HCl monohydrate and the amount of unidentified material is < 1%.
The FEEDAP, however, is aware that the intake of histamine, a metabolic by‐product of histidine, through fish flesh following microbial spoilage is a serious concern for consumers (EFSA BIOHAZ Panel, 2011). Histamine poisoning from fish flesh has been called ‘scombroid’ poisoning because of the edible fish species (e.g. tuna, mackerel) more liable to histamine formation due to the high content of histidine in their flesh. However, bacterial spoilage may induce histamine formation also in other teleost species, including diverse freshwater species such as trout and carp (Křížek et al., 2011) and catfishes (Widjaja et al., 2010). Commission Regulation (EC) No 2073/2005 sets a maximum limit of 200 mg histamine/kg flesh for sea fishery products (raw fish at the point of the first sale) of fish species associated with a high amount of histidine, in particular fish species of the families: Scombridae, Clupeidae, Engraulidae, Coryfenidae, Pomatomidae and Scombresosidae.33 The FEEDAP Panel considers that histamine food poisoning is mainly associated with the consumption of fish.
The Applicant provided two literature searches described and assessed in a previous opinion (EFSA FEEDAP Panel, 2019). The scarce evidence suggests that increasing dietary levels of histidine resulted in increased concentrations of histidine in edible tissues of food‐producing animals. Although histidine is a precursor of histamine, the main factors influencing histamine formation in fish are storage time, temperature, pH, hygienic conditions (e.g. bacterial contamination) or starter cultures of fermented foods, which have been reviewed in previous publications (EFSA BIOHAZ Panel, 2011; FAO, 2013, 2018; Technical report EFSA, 2017).
As pointed out by FAO (2018), ‘the available evidence highlights that under appropriate time × temperature control, and within the sensory shelf‐life of the product, histamine development in Salmonidae to the levels that cause scombroid fish poisoning is unlikely to occur’.
In view of the above, the FEEDAP Panel considers that supplementing the diets of salmonids with histidine to cover the requirements is unlikely to result in the increase of histamine formation, provided that appropriate handling and storage of fish are ensured. Although there is no evidence from other aquaculture species, the Panel considers that the above conclusions can be extrapolated to other commonly farmed fish. For fish species associated with high levels of histidine in flesh,34 the Panel notes that supplemental histidine may increase histidine concentration in fish flesh and the possibility to have higher levels of histamine in fish flesh following unproper storage. However, there are limits established for histamine to protect the consumer, in particular for Scombroid fish species.
In the absence of histamine poisoning records associated with raw mammal or poultry edible tissues and products, the FEEDAP Panel considers it unlikely that supplementation of feed with histidine to cover animal requirements will increase the risk of histamine poisoning upon consumption of such raw edible tissues and products from mammals and birds, provided that appropriate handling and storage are ensured.
3.2.2.1. Conclusions on the safety for the consumer
l‐Histidine HCl monohydrate produced using C. glutamicum KCCM 80179 supplemented at levels appropriate for the requirements of target species and life stage is considered safe for the consumer.
3.2.3. Safety for the user
The analytical data on particle size distribution of the additive indicate that 15–20% (w/w) of the particles had a diameter < 125 μm, and no data on dusting potential were available.
The applicant provided an acute inhalation toxicity test, an eye irritation test, a skin irritation test and a dermal sensitisation test. Those studies tested an l‐histidine monohydrochloride monohydrate produced by fermentation with a different production strain C. glutamicum KCCM 80172 (Batch H20170425, purity 99.48%, year 2017).35 As the purity and physical characteristics of the test item are very similar to the ones of the product under assessment and the production process is the same, the FEEDAP Panel considers that the results of the toxicological studies performed with l‐histidine monohydrochloride monohydrate originating from C. glutamicum KCCM 80172 can be used to support the safety for the user of l‐histidine monochloride monohydrate produced with C. glutamicum KCCM 80179.
Consequently, l‐Histidine HCl monohydrate produced using C. glutamicum KCCM 80179 is considered not irritant to skin, is a mildly irritant to eyes, and it is not a skin sensitiser. The additive does not pose a risk to users by inhalation.
3.2.4. Safety for the environment
The amino acid l‐histidine is a physiological and natural component of animal and plant proteins. It is not excreted as such (but as urea/uric acid and carbon dioxide). The use of l‐histidine in animal nutrition would not lead to any localised increase in its concentration in the environment. The use of amino acids in water for drinking, when given in addition to complete diets with a well‐balanced amino acid profile, would disturb the nitrogen balance and increase nitrogen excretion via urine. The use of l‐histidine HCl monohydrate produced by C. glutamicum KCCM 80179 in animal nutrition is not expected to represent a risk to the environment.
3.3. Efficacy
Efficacy studies are not required for amino acids naturally occurring in proteins of plants and animals. The nutritional role of the amino acid l‐histidine monohydrochloride monohydrate is well established in the scientific literature (NRC, 1994, 1998, 2011, 2012)
In general, the product l‐histidine monohydrochloride monohydrate is considered as an efficacious source of the essential amino acid l‐histidine for non‐ruminant animal species. For the supplemental L‐histidine to be as efficacious in ruminants as in non‐ruminant species, it would require protection against degradation in the rumen.
As l‐histidine is used in food as a flavouring compound, it is expected that it can provide a similar function in feed and no further demonstration of efficacy is necessary.
3.4. Post‐marketing 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 Regulation36 and Good Manufacturing Practice.
4. Conclusions
The use of l‐histidine monohydrochloride monohydrate produced by fermentation using C. glutamicum KCCM 80179 is safe for the target species when used as a nutritional additive to supplement the diet in appropriate amounts to cover the requirements, depending on the species, the physiological state of the animal, the performance level, the environmental conditions, the background amino acid composition of the unsupplemented diet and the status of some essential trace elements such as copper and zinc. This conclusion would also cover the use as a sensory additive.
l‐Histidine HCl monohydrate produced using C. glutamicum KCCM80179 supplemented at levels appropriate for the requirements of species and life stage is considered safe for the consumer.
l‐Histidine HCl monohydrate produced using C. glutamicum KCCM80179 is considered not irritant to skin, is mildly irritant to eyes, and it is not a skin sensitiser. The additive does not pose a risk to users by inhalation.
The use of l‐histidine HCl monohydrate produced by C. glutamicum KCCM80179 in animal nutrition is not expected to represent a risk to the environment.
l‐Histidine HCl monohydrate is considered an efficacious source of the essential amino acid l‐histidine for non‐ruminant animal species. For the supplemental l‐histidine to be as efficacious in ruminants as in non‐ruminant species, it would require protection against degradation in the rumen. It is also considered efficacious as a feed flavouring compound under the proposed conditions of use.
Chronology
| Date | Event |
|---|---|
| 20/06/2018 | Dossier received by EFSA: L‐histidine monhohydrochloride monohydrate feed grade from Corynebacterium glutamicum. Submitted by CJ Europe GmbH. |
| 06/07/2018 | Reception mandate from the European Commission |
| 18/09/2018 | Application validated by EFSA – Start of the scientific assessment |
| 22/10/2018 | Request of supplementary information to the applicant in line with Article 8(1)(2) of Regulation (EC) No 1831/2003 – Scientific assessment suspended. Issues: characterisation of the additive, safety for target species, safety for the consumer and safety for the user. |
| 18/12/2018 | Comments received from Member States |
| 06/12/2018 | Reception of supplementary information from the applicant ‐ Scientific assessment re‐started |
| 20/12/2018 | Reception of the Evaluation report of the European Union Reference Laboratory for Feed Additives |
| 15/01/2019 | Request of supplementary information to the applicant in line with Article 8(1)(2) of Regulation (EC) No 1831/2003 – Scientific assessment suspended Issues: Safety for the target species and for the consumer |
| 11/02/2019 | Reception of supplementary information from the applicant ‐ Scientific assessment re‐started |
| 02/07/2019 | Opinion adopted by the FEEDAP Panel. End of the Scientific assessment |
Abbreviations
- AFC
Scientific Panel on Food Additives, Flavourings, Processing Aids and Materials in Contact with Food
- CAS
Chemical Abstracts Service
- CFU
colony forming unit
- Co‐planar PCB
coplanar dioxin‐like polychlorinated biphenyl
- CV
coefficient of variation
- DM
dry matter
- EINECS
European Inventory of Existing Commercial Chemical Substances
- EURL
European Union Reference Laboratory
- FEEDAP
Panel on Additives and Products or Substances used in Animal Feed
- FLAVIS
EU Flavour Information System
- HPLC‐UV
high‐performance liquid chromatography coupled with ultraviolet detection
- IEC‐VIS
ion exchange chromatography coupled with photometric detection
- IUPAC
International Union of Pure and Applied Chemistry
- KCCM
Korean Culture Centre of Microorganisms
- LOD
limit of detection
- LOQ
limit of quantification
- MIC
minimum inhibitory concentration
- OECD
Organisation for Economic Co‐operation and Development
- PCDD
polychlorinated dibenzodioxin
- PCDF
polychlorinated dibenzofuran
- QPS
qualified presumption of safety
- RH
relative humidity
- Rrec
recovery rate
- RSDip
relative standard deviation for intermediate precision
- RSDr
relative standard deviation for repeatability
- WHO
World Health Organization
- VKM
Norwegian Scientific Committee for Food Safety
Annex A – Evaluation report of the analytical methods submitted in connection with the application for authorisation of l‐histidine monohydrochloride monohydrate produced by fermentation with Corynebacterium glutamicum KCCM 80179
1.
In the current application, authorisation is sought under Article 4 for l‐histidine monohydrochloride monohydrate produced by fermentation with Corynebacterium glutamicum KCCM 80179, under the category/functional group 3(c) ‘nutritional additives’/’amino acids, their salts and analogues’, according to Annex I of Regulation (EC) No 1831/2003. Authorisation is sought for all animal species. According to the Applicant, l‐histidine monohydrochloride monohydrate has a minimum purity (mass fraction) of 98%. The feed additive is intended to be added directly into feedingstuffs or through premixtures. However, the Applicant did not propose any minimum or maximum content of l‐histidine monohydrochloride monohydrate in feedingstuffs.
For the quantification of l‐histidine monohydrochloride monohydrate in the feed additive, the Applicant submitted an in‐house validated analytical method based on reversed‐phase high‐performance liquid chromatography coupled with ultraviolet detection (HPLC‐UV). The Applicant reported in the frame of the validation study a relative standard deviation for repeatability (RSDr) and intermediate precision (RSDip) ranging from 0.1 to 2.1% and a recovery rate (Rrec) ranging from 98 to 102%. Furthermore, the EURL calculated a RSDr of 0.3% from further analytical data presented by the Applicant.
For the quantification of l‐histidine in premixtures and feedingstuffs, the EURL identified the ring‐trial validated Community method (Commission Regulation (EC) No 152/2009) based on ion exchange chromatography coupled with photometric detection (IEC‐VIS). This method, designed for the analysis of amino acids in premixtures and feedingstuffs, does not distinguish between the salts and the amino acid enantiomers. The following performance characteristics were reported for the quantification of total histidine: RSDr ranging from 2.4 to 7.0% and RSDR ranging from 13 to 23%.
Based on the performance characteristics available, the EURL recommends for official control the in‐house validated method based on HPLC‐UV to quantify l‐histidine monohydrochloride monohydrate in the feed additive and the ring‐trial validated Community method based on IEC‐VIS to quantify histidine in premixtures and feedingstuffs.
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, as last amended by Regulation (EU) 2015/1761) is not considered necessary.
Suggested citation: EFSA FEEDAP Panel (EFSA Panel on Additives and Products or Substances used in Animal Feed) , Bampidis V, Azimonti G, de Lourdes Bastos M, Christensen H, Dusemund B, Kouba M, Kos Durjava M, López‐Alonso M, López Puente S, Marcon F, Mayo B, Pechová A, Petkova M, Sanz Y, Villa RE, Woutersen R, Costa L, Cubadda F, Dierick N, Flachowsky G, Mantovani A, Wallace RJ, Manini P, Tarrés‐Call J and Ramos F, 2019. Scientific Opinion on the safety and efficacy of l‐histidine monohydrochloride monohydrate produced using Corynebacterium glutamicum KCCM 80179 for all animal species. EFSA Journal 2019;17(7):5784, 14 pp. 10.2903/j.efsa.2019.5784
Requestor: European Commission
Question number: EFSA‐Q‐2018‐00547
Panel members: Giovanna Azimonti, Vasileios Bampidis, Maria de Lourdes Bastos, Henrik Christensen, Birgit Dusemund, Maryline Kouba, Mojca Kos Durjava, Marta López‐Alonso, Secundino López Puente, Francesca Marcon, Baltasar Mayo, Alena Pechová, Mariana Petkova, Fernando Ramos, Yolanda Sanz, Roberto Edoardo Villa and Ruud Woutersen.
Acknowledgements: The Panel wishes to thank the following for the support provided to this scientific output: Elisa Pettenati, Kostantinos Sofianidis.
Legal notice: Relevant information or parts of this scientific output have been blackened in accordance with the European Commission decision on the confidentiality requests formulated by the applicant. The full output has been shared with the European Commission, EU Member States and the applicant.
Adopted: 2 July 2019
Notes
Regulation (EC) No 1831/2003 of the European Parliament and of the Council of 22 September 2003 on additives for use in animal nutrition. OJ L 268, 18.10.2003, p. 29.
Welding GmbH & Co. KG. Esplanase 39, 20354 Hamburg, Germany.
Commission Regulation (EC) No 244/2007 of 7 March 2007 concerning the authorisation of L‐histidine monohydrochloride monohydrate as feed additive. OJ L 73/6, 13.3.2007, p. 2.
Commission List of the authorised additives in feedingstuffs published in application of Article 9th of Council Directive 70/524/EEC concerning additives in feedingstuffs (2004/C 50/01). OJ C 50/1 25.2.2004.
Regulation (EU) No 609/2013 of the European Parliament and of the Council of 12 June 2013 on food intended for infants and young children, food for special medical purposes, and total diet replacement for weight control and repealing Council Directive 92/52/EEC, Commission Directives 96/8/EC, 1999/21/EC, 2006/125/EC and 2006/141/EC, Directive 2009/39/EC of the European Parliament and of the Council and Commission Regulations (EC) No 41/2009 and (EC) No 953/2009, OJ L 181, 29.6.2013, p.35.
Commission Decision of 9 February 2006 amending Decision 96/335/EC establishing an inventory and a common nomenclature of ingredients employed in cosmetic products. OJ L 97, 5.4.2006, pp. 1–528.
Commission Regulation (EU) No 37/2010 of 22 December 2009 on pharmacologically active substances and their classification regarding maximum residue limits in foodstuffs of animal origin. OJ L 15, 20.1.2010, p. 1.
Regulation (EC) No 470/2009 of the European Parliament and of the Council of 6 May 2009 laying down Community procedures for the establishment of residue limits of pharmacologically active substances in foodstuffs of animal origin, repealing Council Regulation (EEC) No 2377/90 and amending Directive 2001/82/EC of the European Parliament and of the Council and Regulation (EC) No 726/2004 of the European Parliament and of the Council. OL L 152, 16.6.2009, p. 11.
FEED dossier reference: FAD‐2018‐0040.
The full report is available on the EURL website: https://ec.europa.eu/jrc/sites/jrcsh/files/finrep-fad-2018-0040-histidine.pdf
Commission Regulation (EC) No 429/2008 of 25 April 2008 on detailed rules for the implementation of Regulation (EC) No 1831/2003 of the European Parliament and of the Council as regards the preparation and the presentation of applications and the assessment and the authorisation of feed additives. OJ L 133, 22.5.2008, p. 1.
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Technical dossier/Section II.1.3 and annex II.1.1; and supplementary information November 2018/02 SIN CJ L‐HIS 80179, reply to question 1.
Technical dossier/Section II/Annex II.1.4. Analytical method for histidine was HPLC with C18 column using potassium phosphate buffer as mobile phase.
Technical dossier/Section II/Annex II.1.2.
Technical dossier/Section II/Annex II.1.2. LOD (in mg/kg) were 1 for lead; 0.5 for cadmium and arsenic; and 0.1 for mercury.
Technical dossier/Section II/Annex II.1.2. LOD (in µg/kg) was 0.05 for PCDD/F and 0.13 for Co‐PCBs.
Technical dossier/Section II/Annex II.1.2. LOD (in µg/kg) was 0.1 for aflatoxins (B1, B2, G1, G2) and ochratoxin A; 1.5 for zearalenone; 0.5 for deoxynivalenol and 5 for fumonisins B1 and B2.
Technical dossier/Section II/Annex II.1.3. LOD (in µg/kg) was 0.09‐0.15 for nitrofurans and 0.11‐0.16 for nitrofuran metabolites.
Technical dossier/Section II/Annex II.1.4. LOD (in µg/kg) was 0.5‐8.
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Technical dossier/Section II.2.2.1 and Annex II.3.1.
Technical dossier/Section II.2.2.1/Annex II.1.5.
Technical dossier/Section II/Annexes II.4.1, II.4.3, II.4.4., II.4.2 and II.4.5.
Technical dossier/Supplementary information December 2018/Annex Sin 2 CJ.
Technical dossier/Section II/II.5.1.
Technical dossier/Section II.4.1.3.
Commission Regulation (EC) No 2073/2005 of 15 November 2005 on microbiological criteria for foodstuffs. OJ 22.12.2005, L 338/1 26 pp.
According to the Commission Regulation (EC) No 2073/2005, in particular fish species of the families Scombridae, Clupeidae, Engraulidae, Coryfenidae, Pomatomidae and Scombresosidae.
FAD‐2017‐0052/Technical dossier/Section III/Annex III.3.07.
Regulation (EC) No 183/2005 of the European Parliament and of the Council of 12 January 2005 laying down requirements for feed hygiene. OJ L 35, 8.2.2005, p. 1.
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