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EFSA Journal 2012;10(5):2676

Suggested citation: EFSA Panel on Additives and Products or Substances used in Animal Feed (FEEDAP); Scientific
Opinion on the safety and efficacy of L-carnitine and L-carnitine L-tartrate as feed additives for all animal species based on a
dossier submitted by Lonza Benelux BV. EFSA Journal 2012;10(5):2676. [23 pp.] doi:10.2903/j.efsa.2012.2676. Available
online: www.efsa.europa.eu/efsajournal

European Food Safety Authority, 2012
SCIENTIFIC OPINION
Scientific Opinion on the safety and efficacy of L-carnitine and L-carnitine
L-tartrate as feed additives for all animal species based on a dossier
submitted by Lonza Benelux BV
1

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

European Food Safety Authority (EFSA), Parma, Italy
ABSTRACT
L-Carnitine has a key role in the metabolism of fatty acids as a substrate for the reversible acetylation of
coenzyme A and as a carrier for the transport of long-chain fatty acids from cytosol across the inner
mitochondrial membrane. L-Carnitine and L-carnitine L-tartrate administered via feed or water for drinking are
considered safe for the target species. The additives appear to have a wide margin of safety (> 10) at the levels
typically used in feed (1050 mg/kg feed). Very little information is available on the toxicology of L-carnitine.
Nevertheless, based on residue data obtained from multi-fold doses of the typical use levels, the FEEDAP Panel
concluded that typical supplementation of feed with L-carnitine or L-carnitine L-tartrate would not substantially
increase human exposure to carnitine from food of animal origin. Therefore, the FEEDAP Panel considers that
the use of L-carnitine and L-carnitine L-tartrate as additives in animal nutrition is safe for the consumer. L-
Carnitine and L-carnitine L-tartrate are not irritant to skin and eyes nor are they skin sensitisers. L-Carnitine and
L-carnitine L-tartrate showed limited dust formation. As inhalation toxicity studies were not available, adverse
effects in the respiratory tract cannot be fully excluded. The use of L-carnitine and L-carnitine L-tartrate in
animal nutrition is not expected to pose a risk to the environment. L-Carnitine and L-carnitine L-tartrate are
regarded as an effective source of L-carnitine in all animal species.
European Food Safety Authority, 2012

KEY WORDS
Nutritional additive, vitamins and provitamins, L-carnitine, L-carnitine L-tartrate, safety

1
On request from the European Commission, Question No EFSA-Q-2011-00251, adopted on 24 April 2012.
2
Panel members: Gabriele Aquilina, Georges Bories, Andrew Chesson, Pier Sandro Cocconcelli, Joop de Knecht, Nol
Albert Dierick, Mikolaj Antoni Gralak, Jrgen Gropp, Ingrid Halle, Christer Hogstrand, Reinhard Kroker, Lubomir Leng,
Secundino Lpez Puente, AnneKatrine Lundebye Haldorsen, Alberto Mantovani, Giovanna Martelli, Mikls Mzes,
Derek Renshaw, Maria Saarela, Kristen Sejrsen and Johannes Westendorf. Correspondence: FEEDAP@efsa.europa.eu
3
Acknowledgement: The Panel wishes to thank the members of the Working Group on Watersoluble Vitamins, including
Annette Schuhmacher, for the preparatory work on this scientific opinion.

L-Carnitine and L-carnitine L-tartrate for all animal species

EFSA Journal 2012;10(5):2676 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 L-carnitine and L-carnitine L-tartrate as additives to feed and water for drinking for all
animal species.
L-Carnitine has a key role in the metabolism of fatty acids as a substrate for the reversible
acetylation of coenzyme A and as a carrier for the transport of long-chain fatty acids from cytosol
across the inner mitochondrial membrane. Adult animals can synthesise carnitine from its precursor
lysine; however, endogenous synthesis might be insufficient in juvenile animals. Nutritional
requirement data for the different animal species and categories have not been established.
Oral administration routes of L-carnitine and L-carnitine L-tartrate via feed or water for drinking are
considered bioequivalent.
L-Carnitine and L-carnitine L-tartrate are safe for the target species. The additives appear to have a
wide margin of safety (> 10) compared at the levels typically used in feed (1050 mg L-carnitine/kg
feed).
Very little information is available on the toxicology of L-carnitine. Nevertheless, based on residue
data obtained from multi-fold doses of the typical use levels, the FEEDAP Panel concluded that
typical supplementation of feed with L-carnitine or L-carnitine L-tartrate would not substantially
increase human exposure to carnitine from food of animal origin. As the absorption rate declines with
increasing L-carnitine intake, the endogenous carnitine pool may not significantly increase. Therefore,
the FEEDAP Panel considers that the use of L-carnitine and L-carnitine L-tartrate as additives in
animal nutrition is safe for the consumer.
L-Carnitine and L-carnitine L-tartrate are not irritant to skin and eyes nor are they skin sensitisers. L-
Carnitine and L-carnitine L-tartrate showed limited dust formation. As inhalation toxicity studies were
not available, adverse effects in the respiratory tract cannot be fully excluded.
Carnitine is a naturally occurring substance, present in all animals and bacteria and, to a lesser extent,
in some plants. The use of L-carnitine and L-carnitine L-tartrate in animal nutrition is not expected to
pose a risk to the environment.
L-Carnitine and L-carnitine L-tartrate are regarded as effective sources of L-carnitine in all animal
species.
The FEEDAP Panel recommends the use of the specifications for L-carnitine according to PhEur (MG
1339) considering purity, substance-related impurities and other impurities (sulphated ash and heavy
metals).
L-Carnitine and L-carnitine L-tartrate for all animal species

EFSA Journal 2012;10(5):2676 3
TABLE OF CONTENTS
Abstract .................................................................................................................................................... 1
Summary .................................................................................................................................................. 2
Table of contents ...................................................................................................................................... 3
Background .............................................................................................................................................. 4
Terms of reference.................................................................................................................................... 4
Assessment ............................................................................................................................................... 6
1. Introduction ..................................................................................................................................... 6
2. Characterisation ............................................................................................................................... 6
2.1. L-Carnitine ................................................................................................................................... 6
2.2. L-Carnitine L-tartrate ................................................................................................................... 7
2.3. Manufacturing process ................................................................................................................ 8
2.3.1. L-Carnitine .......................................................................................................................... 8
2.3.2. L-Carnitine L-tartrate .......................................................................................................... 8
2.4. Stability and homogeneity ........................................................................................................... 8
2.4.1. L-Carnitine .......................................................................................................................... 8
2.4.2. L-Carnitine L-tartrate .......................................................................................................... 9
2.4.3. Homogeneity ...................................................................................................................... 9
2.5. Physico-chemical incompatibilities in feed ................................................................................. 9
2.6. Conditions of use ......................................................................................................................... 9
2.7. Evaluation of the analytical methods by the European Union Reference Laboratory (EURL) . 10
3. Safety ............................................................................................................................................. 10
3.1. Safety for the target species ....................................................................................................... 10
3.1.1. Conclusions on the safety for target species ..................................................................... 10
3.2. Safety for the consumer ............................................................................................................. 10
3.2.1. Absorption, distribution, metabolism, residues and excretion .......................................... 10
3.2.2. Toxicological studies ........................................................................................................ 11
3.2.3. Assessment of consumer safety ........................................................................................ 11
3.2.4. Conclusions on the safety for the consumer ..................................................................... 11
3.3. Safety for the user ...................................................................................................................... 12
3.3.1. Effects on the respiratory system ...................................................................................... 12
3.3.2. Effects on the eyes and skin ............................................................................................. 12
3.3.3. Conclusions on the safety for the user .............................................................................. 13
3.4. Safety for the environment ........................................................................................................ 13
4. Efficacy .......................................................................................................................................... 13
5. Post-market monitoring ................................................................................................................. 13
Conclusions and recommendations ........................................................................................................ 14
Documentation provided to EFSA ......................................................................................................... 14
References .............................................................................................................................................. 14
Appendices ............................................................................................................................................. 19
Appendix A ............................................................................................................................................ 19
Appendix B ............................................................................................................................................ 21
References .............................................................................................................................................. 22
L-Carnitine and L-carnitine L-tartrate for all animal species

EFSA Journal 2012;10(5):2676 4
BACKGROUND
Regulation (EC) No 1831/2003
4
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 also 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
time limit or pursuant to Directive 82/471/EEC.
The European Commission received a request from the company Lonza Benelux BV
5
for (i)
authorisation of a new use (i.e. use in water for drinking), and (ii) re-evaluation of authorisation of the
product carnitine in the form of L-carnitine and L-carnitine L-tartrate, 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) (re-evaluation
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
2011.
7

Carnitine in the form of L-carnitine and L-carnitine L-tartrate has been authorised without time limit
under Council Directive 70/524/EEC
8
for its use for all animal species as a nutritional additive.
The Panel on Food Additives, Flavourings, Processing Aids and Materials in Contact with Food
(AFC) expressed an opinion related to L-carnitine L-tartrate for use in foods for particular nutritional
purposes (EFSA, 2003). 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, 2011).
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 animals, consumer, user and the environment and the efficacy of carnitine in the
form of L-carnitine and L-carnitine L-tartrate, when used under the conditions described in Table 1.

4
Regulation (EC) 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.
5
Lonza Benelux BV, Aluminiumstraat 1, 4800 AL Breda, NL.
6
EFSA Dossier reference: FAD20100144.
7
A new mandate was received in EFSA in March 2011.
8
Commission list of the authorised additives in feedingstuffs published in application of Article 9t (b) of Council Directive
70/524/EEC concerning additives in feedingstuffs (2004/C 50/01). OJ C 50, 25.2.2004, p. 1.
L-Carnitine and L-carnitine L-tartrate for all animal species

EFSA Journal 2012;10(5):2676 5
Table 1: Description and conditions of use of the additive as proposed by the applicant
Additive
L-Carnitine
L-Carnitine L-tartrate
Registration number/EC No/No
(if appropriate)
-
Category of additive 3. Nutritional additives
Functional group(s) of additive
A. Vitamins, pro-vitamins and chemically well defined
substances having a similar effect.


Description
Composition, description
Chemical
formula
Purity criteria
(if appropriate)
Method of analysis
(if appropriate)
L-Carnitine C
7
H
15
NO
3
min. 97.0 %
Titration with
hydrochloric acid
L -Carnitine L-tartrate C
18
H
36
N
2
O
12
min. 97.0 %
Backtitration with
hydrochloric acid
and sodium
hydroxide


Trade name (if appropriate) Not appropriate
Name of the holder of authorisation
(if appropriate)
Not appropriate


Conditions of use
Species or category
of animal
Maximum
Age
Minimum content
Maximum
content Withdrawal
period
(if appropriate)
mg kg
-1
of complete feedingstuffs,
supplementary feed (based on end feed)
and in water*
All animal species
and categories
- - - -
Other provisions and additional requirements for the labelling
Specific conditions or restrictions for use (if
appropriate)
Not applicable
Specific conditions or restrictions for
handling (if appropriate)
Please refer to MSDS
Post market monitoring
(if appropriate)
Not applicable
Specific conditions for use in
complementary feedingstuffs or water
(if appropriate)
Not applicable


Maximum Residue Limit (MRL) (if appropriate)
Marker residue
Species or category of
animal
Target tissue(s)
or food products
Maximum
content in tissues
- - - -
L-Carnitine and L-carnitine L-tartrate for all animal species

EFSA Journal 2012;10(5):2676 6
ASSESSMENT
This opinion is based in part on data provided by a single company involved in the production of
carnitine in the form of L-carnitine and L-carnitine L-tartrate. It should be recognised that these data
from a single applicant cover only a fraction of existing additives containing L-carnitine and L-
carnitine L-tartrate. The composition of the additives is not the subject of the application. The Panel
has sought to use the data provided, together with data from other sources, to deliver an opinion.
1. Introduction
L-Carnitine (-hydroxy--trimethyl aminobutyric acid), has a key role in the metabolism of fatty acids
(energy) as a substrate for the reversible acetylation of coenzyme A and as a carrier for the transport
of long-chain fatty acids from cytosol across the inner mitochondrial membrane. It is therefore
important for the energy metabolism of the animal. L-Carnitine is a dispensable compound as adult
animals can synthesize carnitine from its precursor lysine; however, endogenous synthesis might be
insufficient in juvenile animals (for details see Annex B).
Carnitine in the form of L-carnitine and L-carnitine L-tartrate 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 its use in all animal species as a nutritional additive.
The applicant has asked for a re-evaluation of the use of L-carnitine and L-carnitine L-tartrate as
additives to feed and for a new use in water for drinking. The substances are intended as nutritional
additives under the functional group vitamins, pro-vitamins and chemically well-defined substances
having similar effects, for all animal species and categories.
L-Carnitine, L-carnitine hydrochloride and L-carnitine L-tartrate are authorised for addition for specific
nutritional purposes in foods for particular nutritional uses (Regulation (EC) No 953/2009),
10
to
processed cereal-based foods and baby foods for infants and young children (Directive 2006/125/EC,
Annex IV)
11
and to infant formulae and follow-on formulae when reconstituted as instructed by the
manufacturer (Directive 2006/141/EC, Annex III).
12
Carnitine is also listed as a pharmacologically
active substance in veterinary medicinal products (Commission Regulation (EC) No 37/2010),
13
and it
is not subject to maximum residue limits when used in food-producing animals.
Levocarnitine is described in the European Pharmacopeia (PhEur) in Monograph (MG) 1339.
2. Characterisation
2.1. L-Carnitine
L-Carnitine (IUPAC name: (3R)-3-hydroxy-4-(trimethylazaniumyl) butanoate; synonym: -hydroxy--
trimethyl aminobutyric acid) is identified by the CAS (Chemical Abstracts Service) number 541-15-1
and the EINECS (European Inventory of Existing Chemical Substances) number 208-768-0. The
structural formula of L-carnitine is shown in Figure 1.

9
Commission list of the authorised additives in feedingstuffs published in application of Article 9t (b) of Council Directive
70/524/EEC concerning additives in feedingstuffs (2004/C 50/01). OJ C 50, 25.2.2004, p. 1.
10
Commission Regulation (EC) 953/2009 of 13 October 2009 on substances that may added for specific nutritional purposes
in foods for particular nutritional uses. OJ L 269, 14.10.2009, p. 9.
11
Commission Directive 2006/125/EC of 5 December 2006 on processed cerealbased foods and babyfoods for infants and
young children. OJ L 339 6.12.2006, p. 16.
12
Commission Directive 2006/141/EC of 22 December 2006 on infant formulae and follow-on formulae and amending
Directive 1999/21/EC. OJ L 401 30.12.2006, p. 1.
13
Commission Regulation (EU) 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.
L-Carnitine and L-carnitine L-tartrate for all animal species

EFSA Journal 2012;10(5):2676 7

Figure 1: Structural formula of L-carnitine.
The molecular formula of L-carnitine is C
7
H
15
NO
3
and its molecular weight is 161.20. It has a melting
point of 180197 C, shows a bulk density of approximately 0.64 g/cm
3
and is readily soluble in water
(12.7 g/mL at 2025 C according to different sources), soluble in ethanol and methanol and slightly
soluble in acetone.
L-Carnitine is a white to beige odourless hygroscopic powder. The analysis of five batches of the
product showed an average content of 100.1 0.3 % L-carnitine and 0.3 0.2 % water,
14
which meets
the minimum specifications of PhEur MG 1339 (98 % L-carnitine in the anhydrous substance). The
main impurity, 4-hydroxycrotonic acid, was below 0.01 % (0.0010.009 %). The amount of
substance-related impurities (impurities AD, according to PhEur) was below 0.05 % and complies
with the thresholds of PhEur, as demonstrated by the analysis of five batches
15
. Data for residual
solvents indicated that VICH (International Cooperation on Harmonisation of Technical Requirements
for Registration of Veterinary Medicinal Products) thresholds for class 2 and 3 are not exceeded.
Production batches are routinely screened for chemical and microbial contamination and impurities.
From that routine survey the following values were reported as typical: mercury < 0.1 mg/kg,
cadmium < 0.25 mg/kg, lead < 1.5 mg/kg, arsenic < 0.04 mg/kg, total aerobic microorganisms < 50
colony-forming units (CFUs)/g.
Six batches of L-carnitine were analysed for particle size distribution determined by laser diffraction.
The particle size distribution varied considerably from batch to batch: the fraction of particles below
50 m was in the range 10.949.7 % (v/v). The additive (three batches) showed a low dusting
potential of 0.060.25 g/m
3
in the StauberHeubach test.
16

2.2. L-Carnitine L-tartrate
L-Carnitine L-tartrate [IUPAC name 1-propanaminium, 3-carboxy-2-hydroxy-N,N,N-trimethyl-, (R)-,
salt with (R-(R*,R*)-2,3-dihydroxybutanedioic acid (2:1); synonyms: -hydroxy--trimethyl
aminobutyrate, L-tartrate, L-carnitine L-tartrate (2:1)] is identified by CAS number 36687-82-8. The
structural formula of L-carnitine L-tartrate is shown in Figure 2.

Figure 2: Structural formula of L-carnitine L-tartrate.
The molecular formula of L-carnitine L-tartrate is C
18
H
36
N
2
O
12
and its molecular weight is 472.49. It
has melting point of approximately 170 C (with decomposition), shows a bulk density of
approximately 0.70 g/cm
3
and is highly soluble in water (> 1 g/mL at 20 C).

14
Technical dossier/Section II.
15
Technical dossier/Supplementary information February 2012.
16
Technical dossier/Section II and supplementary information October 2011/Annexes 2.33 and 2.34 and supplementary
information February 2012/Annexes 2.45 and 2.48.
L-Carnitine and L-carnitine L-tartrate for all animal species

EFSA Journal 2012;10(5):2676 8
L-Carnitine L-tartrate is a crystalline white powder. The additive contains by specification at least
97.0 % L-carnitine L-tartrate, which corresponds to at least 67 % L-carnitine and less than 0.5 % water.
Analysis of five batches of the product showed an average content of L-carnitine, L-tartaric acid and
water of 68.5 0.7 %, 31.5 0.2 % and 0.2 0.1 %, respectively.
17
The main impurity, 4-
hydroxycrotonic acid, was below 0.02 % (< 0.010.02 %) and total related impurities were below
0.05 % (determined by HPLC). Data for residual solvents indicated that VICH thresholds for class 2
and 3 are not exceeded.
Production batches are routinely screened for chemical and microbial contamination and impurities.
The following values were reported as typical: mercury < 0.005 mg/kg, cadmium < 0.02 mg/kg, lead
< 0.03 mg/kg, arsenic < 0.04 mg/kg, total aerobic microorganisms < 50 CFUs/g.
Three batches of L-carnitine L-tartrate were analysed for particle size distribution determined by laser
diffraction. The fraction of particles below 50 m was in the range 2.83.4 % (v/v). The substance
showed a dusting potential of 0.19 g/m
3
.
18

2.3. Manufacturing process
2.3.1. L-Carnitine
L-Carnitine is produced by a two-step chemical synthesis process. In the first step, a solution of ethyl
4-chloroacetoacetate in ethanol is hydrogenated using a chiral catalyst. In the second step, ethyl (R)-4-
chloro-3-hydroxybutyrate is aminated with trimethylamine and hydrolysed with aqueous sodium
hydroxide, giving L-carnitine. The obtained aqueous L-carnitine solution is electrodialysed and re-
crystallised in an organic solvent and then dried. The applicant provided a flow chart of the synthetic
process.
A formulated product containing about 50 % L-carnitine is produced by the addition of a binder with a
high water absorption capacity such as a silicon source.
2.3.2. L-Carnitine L-tartrate
L-Tartaric acid in an organic solvent is added to an aqueous solution of L-carnitine. The solution is
cooled and centrifuged. The resultant L-carnitine L-tartrate crystals are then dried.
2.4. Stability and homogeneity
2.4.1. L-Carnitine
2.4.1.1. Shelf life
L-Carnitine (one batch, stored in double polyethylene bags) was demonstrated to have a shelf life of 36
months at 25 C. Shelf life under accelerated conditions at 40 C was shown for one batch of the
product to be six months. The shelf life of a formulated product containing 50 % L-carnitine (one
batch, stored in polyethylene bags at 25 C) did not differ from that of the active substance.
19

2.4.1.2. Stability of the additive when added to premixtures, feed and water for drinking
Three batches of a vitaminmineral premixture for piglets containing also choline chloride
20

supplemented with 8 000 mg L-carnitine per kilogram (from an additive containing 50 % L-carnitine)

17
Technical dossier/Section II and Supplementary information October 2011.
18
Technical dossier/Section II and Supplementary information October 2011/Annexes 2.35 and 2.36.
19
Technical dossier/Section II.
20
Technical dossier/Section II/Annexes 2.10 and 2.12.
L-Carnitine and L-carnitine L-tartrate for all animal species

EFSA Journal 2012;10(5):2676 9
showed no reduction in the L-carnitine content when kept in polyethylene bags at 25 C for up to six
months.
21

Stability of L-carnitine was also investigated in mash and pelleted feed for piglets (three batches each)
when added at 50 mg L-carnitine per kilogram complete feed. Feed processing did not affect the
concentration of L-carnitine in feed. After storage for three months at 25 C no reduction in the content
of L-carnitine was observed.
22

No reduction in the concentration of L-carnitine (three batches) was seen in water for drinking when
added at 1 g/L and stored for 24 hours at 25 C.
23

2.4.2. L-Carnitine L-tartrate
2.4.2.1. Shelf life
L-Carnitine L-tartrate (three batches stored in glass vials) was demonstrated to have a shelf life of 48
months at 25 2 C.
24

2.4.2.2. Stability of the additive when added to premixtures, feed and drinking water
The applicant provided no information on the stability of L-carnitine L-tartrate in premixtures and
complete feed, but stated that the stability of L-carnitine L-tartrate is similar to that of L-carnitine. The
FEEDAP Panel is therefore not in the position to draw conclusions on the stability of the additive L-
carnitine L-tartrate in premixtures and complete feed. However, even when hydrolysis occurs the L-
carnitine component is likely to be stable.
No reduction in the concentration of L-carnitine (intended value 1 g/L) was seen in water for drinking
when L-carnitine Ltartratee (three batches) was added at 1.4 g/L and stored for 24 hours at 25 C.
25

2.4.3. Homogeneity
The capacity for homogeneous distribution was examined in feed for L-carnitine (intended
concentration 50 mg/kg) and in a premixture and in feed for L-carnitine L-tartrate (intended
concentration 5 000 and 60 mg L-carnitine/kg, respectively) by analysis in triplicate of five samples
each. The coefficient of variation was 5.1 %, 3.4 % and 7.0 %, respectively.
26

L-Carnitine and L-carnitine L-tartrate are highly soluble in water. Therefore, homogeneity in water for
drinking need not be demonstrated.
2.5. Physico-chemical incompatibilities in feed
No physico-chemical incompatibilities or interactions have been reported between L-carnitine/L-
carnitine L-tartrate and feed materials, carriers, other approved additives or medicinal products when
added to premixtures and feed. No such incompatibilities or interactions were expected.
2.6. Conditions of use
L-carnitine and L-carnitine L-tartrate are intended for use via feed (premixtures, complete or
complementary feed) and water for drinking in all animal species and categories without a maximum
content or withdrawal period.

21
Technical dossier/Section II/Annex 2.9.
22
Technical dossier/Section II/Annex 2.13.
23
Technical dossier/Section II/Annex 2.14 and supplementary information October 2011.
24
Technical dossier/Section II.
25
Technical dossier/Section II/Supplementary information October 2011/Annex 2.38.
26
Technical dossier/Section II/Annex 2.15 and supplementary information October 2011/Annexes 3.39 and 2.40.
L-Carnitine and L-carnitine L-tartrate for all animal species

EFSA Journal 2012;10(5):2676 10
Two additives containing L-carnitine are mentioned in the dossier, one containing at least 97 % L-
carnitine and intended for use in milk replacers, wet feed formulation and water for drinking and
another containing about 50 % L-carnitine intended for supplementation of other solid feed.
2.7. 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 L-carnitine and
L-carnitine L-tartrate in animal feed. The executive summary of the EURL report can be found in
Appendix A.
3. Safety
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 FEEDAP considers is
the case for L-carnitine.
3.1. Safety for the target species
L-Carnitine is well tolerated by humans and all animal species, possibly because of its decreasing
bioavailability with increasing dietary L-carnitine concentrations. In humans the bioavailability of L-
carnitine declines to about 16 % and 5 % after oral administration of a single dose of 2 or 6 g L-
carnitine (Harper et al., 1988).
In studies with Japanese quail (Coturnix coturnix subsp. japonica), a dietary concentration of
8 000 mg L-carnitine/kg did not cause any adverse effect; also the addition of 8 000 mg D-carnitine/kg,
which competitively inhibits the carrier-mediated transport and synthesis of L-carnitine, did not affect
the zootechnical parameters of the quail (Kliemant, 2000). In horses no adverse effects could be
observed when 5, 10 or 20 g of L-carnitine was administered via gavage every three days, or 12 g
daily via the feed; the increase from 5 to 20 g L-carnitine/day increased free and total carnitine in the
plasma by only 25 % and 33 %, respectively (Schnitger, 2003). Doses of 1060 g per horse (as two or
three doses per day) were without adverse effects in thoroughbred horses (Foster et al., 1988; Harris et
al., 1995). Studies with growing pigs and early weaned piglets suggest that dietary concentrations of
about 5006 000 mg L-carnitine/kg are well tolerated (Newton and Haydon, 1988, 1989; Owen et al.,
1994, 1996, 2001a; Ghler, 2002; Heo et al., 2000).
3.1.1. Conclusions on the safety for target species
Based on the literature it can be concluded that L-carnitine and L-carnitine L-tartrate are safe for the
target species. The additives appear to have a wide margin of safety (> 10) at the levels typically used
in feed (between 10 and 50 mg L-carnitine/kg feed).
3.2. Safety for the consumer
3.2.1. Absorption, distribution, metabolism, residues and excretion
After ingestion L-carnitine is absorbed in the small intestine via a sodium-dependent transport system
(low carnitine concentration) or at pharmacological doses of carnitine via passive diffusion as free
carnitine and short-chain carnitine esters (Shaw et al., 1983; Gudjonsson et al., 1985; Hamilton et al.,
1986; Li et al., 1990; Marciani et al., 1991). The bioavailability of dietary L-carnitine is about 54
87 % in humans and dependent on the dietary L-carnitine content (Rebouche and Chenard, 1991;
Rebouche, 2004). The major portion of all carnitine in the body is found in the skeletal muscle (see,
for example, Mitchell 1978). Studies with labelled L-carnitine suggest that carnitine is usually largely
not metabolised (e.g. Lindstedt and Lindstedt, 1961; Yue and Fritz, 1962; Brooks and MacIntosh,
1975; Bremer, 1983); however, carnitine might be converted, for example to -methyl choline, during
stress and disease or depending on the dietary or physiological conditions (Mehlman et al., 1969;
L-Carnitine and L-carnitine L-tartrate for all animal species

EFSA Journal 2012;10(5):2676 11
Mitchell, 1978). Carnitine is excreted mainly via the kidney with a highly efficient tubular
reabsorption of about 9099 % at normal dietary intakes and normal plasma and tissue concentrations
(Harper et al., 1988; Rebouche 2004); only 2 % of the ingested L-carnitine is voided in the faeces
(Carroll et al., 1981).
In studies with Japanese quail fed a wheat/soybean-based diet without or with an addition of 2 000 mg
L-carnitine per kilogram feed for 28 days, the total carnitine concentration (free carnitine, short-chain
acyl-carnitine and long-chain acyl-carnitine) could be increased (P < 0.05) from 56 to 110 mg/kg
skeletal muscle (M. pectoralis), from 52 to 110 mg/kg cardiac muscle and from 83 to 200 mg/kg liver
(all values are fresh matter; Jger, 2000), i.e. the L-carnitine concentration had at least doubled in each
of the tissues as a result of dietary carnitine supplementation. Comparable results were obtained in an
experiment with early weaned piglets fed 0 or 632 mg L-carnitine/kg feed (wheat/soybean meal). After
4956 days of feeding, the carnitine concentration significantly increased from 273 to 463 mg/kg
skeletal muscle, from 100 to 169 mg/kg cardiac muscle, from 56 to 99 mg/kg kidney and from 35 to
62 mg/kg liver of the control and carnitine-supplemented group, respectively (all values as fresh
matter; Ghler, 2002).
3.2.2. Toxicological studies
The available toxicity data on L-carnitine L-tartrate have been assessed by the Panel on Food
Additives, Flavourings, Processing Aids and Materials in Contact with Food (EFSA, 2003). A dose of
3 g L-carnitine L-tartrate given daily to volunteers for three weeks was tolerated.
An acute oral gavage toxicity test with L-carnitine L-tartrate in rats showed no effects at a dose of
5000 mg/kg body weight.
A gene mutation test was performed with L-carnitine L-tartrate in Salmonella Typhimurium strains TA
1535, TA 1537, TA 1538, TA 98 and TA 100. It was tested at concentrations of 1.6, 8, 40, 200, 1 000
and 5000 g per plate, without and with metabolic activation using rat liver S9 mix. There were two
independent experiments. There was no evidence of gene mutation (EFSA, 2003).
3.2.3. Assessment of consumer safety
No acceptable daily intake or tolerable upper intake level has been established for L-carnitine or L-
carnitine L-tartrate.
L-Carnitine is present in the human diet in a variety of food sources. The richest sources of dietary L-
carnitine (proximate L-carnitine content in brackets) are animal products, such as mutton (210 mg/kg),
beef (60 mg/kg), pork (27 mg/kg) and fish (35 mg/kg). Lower levels of L-carnitine are found in dairy
products (2 mg/l cows milk), whereas most fruits and vegetables (05 mg/kg) contain minimal
amounts of L-carnitine (for details see Mitchell, 1978; Rebouche 1984; Gtz, 1989; Demarquoy et al.,
2004; Knttel-Gustavsen and Harmeyer, 2007).
Human dietary L-carnitine intake ranges from < 0.2 to about 2.4 mg/kg body weight per day
(Rebouche, 2004).
3.2.4. Conclusions on the safety for the consumer
Based on residue data described under section 3.2.1, the FEEDAP Panel concluded that supplementing
feed with L-carnitine or L-carnitine L-tartrate at practical use levels between 10 and 50 mg L-
carnitine/kg feed would not substantially increase human exposure to carnitine from food of animal
origin. As the absorption rate declines with increasing L-carnitine intake, the endogenous carnitine
pool may not significantly increase. Therefore, the FEEDAP Panel considers the use of L-carnitine and
L-carnitine L-tartrate as additives in animal nutrition to be safe for consumers.
L-Carnitine and L-carnitine L-tartrate for all animal species

EFSA Journal 2012;10(5):2676 12
3.3. Safety for the user
3.3.1. Effects on the respiratory system
L-Carnitine showed between 10 % and 49 % of particles (v/v) with a diameter < 50 m; the
corresponding figure for L-carnitine L-tartrate was 3 %. However, the dusting potential for both
additives was low ( 0.25 g/m
3
). In the absence of data on the inhalation toxicity of L-carnitine and L-
carnitine L-tartrate, inhalation of dust is considered to be potentially hazardous.
3.3.2. Effects on the eyes and skin
3.3.2.1. L-Carnitine
In a skin irritation study [OECD (Organization for Economic Cooperation and Development)
guideline 404) the back skin of three New Zealand White rabbits was exposed to 0.5 g L-carnitine for
4 hours.
27
As slight erythema was observed at 1 hour but disappeared within two days, L-carnitine is
regarded as non-irritant to skin (Regulation (EC) No 1272/2008).
28

For an acute eye irritation/corrosion study (OECD guideline 405) 0.1 mL of L-carnitine (67 mg) was
instilled into one of the eyes of three New Zealand White rabbits.
29
Moderate conjunctival irritation
(redness, chemosis and discharge) and minimal to moderate conjunctival irritation was observed in
two treated eyes, in one case at 24 hours and in the other case at 48 hours. The results suggest that L-
carnitine can be regarded as non-irritant to eyes (Regulation (EC) No 1272/2008).
In a Buehler test for skin sensitisation (OECD guideline 406) 20 albino guinea pigs were treated with
75 % L-carnitine (w/w) in distilled water during the induction period, and 50 % and 75 % L-carnitine
(w/w) in distilled water during the challenge phase; 10 guinea pigs served as control group.
30
No skin
reaction was observed after topical challenge. The results indicate that L-carnitine is not a skin
sensitiser.
3.3.2.2. L-Carnitine L-tartrate
In a dermal toxicity study (OECD guideline 402) L-carnitine L-tartrate was administered by a single
dermal application (2000 mg/kg body weight) to five rats of each sex for 24 hours.
31
No abnormalities
were observed at macroscopic post-mortem examination (day 15). The dermal LD
50
value of L-
carnitine L-tartrate was > 2 000 mg/kg.
In an acute skin irritation/corrosion study (OECD guideline 404) the skin of three New Zealand White
rabbits was exposed to 0.5 g L-carnitine L-tartrate for four hours.
32
At 1, 24, 48 and 72 hours after
removal no signs of either irritation or corrosion were observed. The results therefore indicate that L-
carnitine L-tartrate is non-irritant.
For an acute eye irritation study (OECD guideline 405) 0.1 mL of L-carnitine L-tartrate (92 mg) was
instilled into one eye of each of three New Zealand White rabbits.
33
Instillation resulted in irritation of
the conjunctivae (redness, chemosis and discharge), which had cleared within 72 hours in two rabbits
and within 7 days in one rabbit. It was concluded that L-carnitine L-tartrate is non-irritant to eyes.

27
Technical dossier/Section III/Reference 3.18.
28
Regulation (EC) 1272/2008 of 16 December 2008 on classification, labelling and packaging of substances and mixtures,
amending and repealing Directive 67/548/EEC and 1999/45/EC, and amending Regulation (EC) No 1907/2006. OJ C 353,
31.12.2008, p. 1.
29
Technical dossier/Section III/Reference 3.19.
30
Technical dossier/Section III/Reference 3.20.
31
Technical dossier/Section III/Reference 3.21.
32
Technical dossier/Section III/Reference 3.22.
33
Technical dossier/Section III/Reference 3.23.
L-Carnitine and L-carnitine L-tartrate for all animal species

EFSA Journal 2012;10(5):2676 13
A local lymph node assay (OECD guideline 429) was performed to study the skin sensitisation
potential of L-carnitine L-tartrate.
34
Three groups of five female CBA strain mice each were treated
with 10 %, 25 % or 50 % L-carnitine L-tartrate, by open application on the ears. The results suggested
that L-carnitine L-tartrate is not a skin sensitiser.
3.3.3. Conclusions on the safety for the user
L-Carnitine and L-carnitine L-tartrate are not irritating to skin and eyes and are not skin sensitisers. L-
Carnitine and L-carnitine L-tartrate in the formulations described show limited dust formation. As no
information is available on inhalation toxicity, adverse effects on the respiratory tract cannot be fully
excluded.
3.4. Safety for the environment
Carnitine is a naturally occurring substance, present in all animals and bacteria and, to a lesser extent,
in some plants. Its use in animal nutrition is not expected to substantially increase the concentration in
the environment. Therefore, any risk to the environment resulting from the use of carnitine in animal
nutrition is not foreseen.
4. Efficacy
According to Regulation (EC) No 429/2008, efficacy studies are not required for vitamins, pro-
vitamins and chemically well-defined substances having similar effects which are already authorised
as feed additives.
Requirement/allowance data that could be used to derive feed supplementation levels are not
established (McDowell, 2002; AWT, 2002). L-Carnitine and L-carnitine L-tartrate are regarded as
effective sources of L-carnitine in all animal species.
Intensive research has been done on the action of L-carnitine in the fatty acid and energy metabolism
of humans and animals; a number of studies also deal with the effect of additional dietary L-carnitine
on endogenous carnitine concentrations and fat and energy utilisation, as well as on the performance
of various livestock animals (e.g. Newton and Haydon, 1986; Gnther, 1989; Musser et al., 1999a,b;
Owen et al., 2001a,b; Rincker et al., 2003; Ramanau et al., 2004, 2005; Birkenfeld et al., 2006; Fischer
et al., 2009) and pets and racing animals (e.g. Dubelaar et al., 1991a,b; Gevaert et al., 1991; Borghijs
and DeWilde, 1992; Pelletier, 1992; Falaschini and Trombetta, 1993), as well as different fish species
(Santulli and DAmelio, 1986; Santulli et al., 1988, 1990; Torrelle et al., 1991; Burtle, 1993; Ji et al.,
1996). However, the carnitine doses stated to be effective for improving the performance varied not
only among the species investigated but also for one species between the research groups. Moreover,
the occurrence of a carnitine effect is often inconsistent, possibly depending on the experimental diets,
particularly the presence of the precursors, age and growth rate of the animal and the general
experimental conditions (e.g. Burtle, 1990; Litz, 1993; Schuhmacher et al., 1993; Schuhmacher and
Gropp, 1998; Ghler, 2002). In studies with piglets and freshly hatched quail L-carnitine appeared to
improve weight gain and feed efficiency only if the metabolic precursors lysine or methionine were
deficient in the diets (Schuhmacher et al., 1993). However, it seems very unlikely that an amino acid-
sparing effect on a molar basis was responsible for these findings.
5. Post-market monitoring
The FEEDAP Panel considers that there is no need for a specific post-market monitoring plan, other
than the requirements established in the Feed Hygiene Regulation
35
and good manufacturing practice.

34
Technical dossier/Section III/Reference 3.24.
35
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.
L-Carnitine and L-carnitine L-tartrate for all animal species

EFSA Journal 2012;10(5):2676 14
CONCLUSIONS AND RECOMMENDATIONS
CONCLUSIONS
Oral administration routes of L-carnitine and L-carnitine L-tartrate via feed or water for drinking are
considered bioequivalent.
L-Carnitine and L-carnitine L-tartrate are safe for the target species. The additives appear to have a
wide margin of safety (> 10) at the levels typically used in feed (1050 mg L-carnitine/kg feed).
Very little information is available on the toxicology of L-carnitine. Nevertheless, based on residue
data obtained from multi-fold doses of the levels typically use, the FEEDAP Panel concluded that
typical supplementation of feed with L-carnitine or L-carnitine L-tartrate would not substantially
increase human exposure to carnitine from food of animal origin. As the absorption rate declines with
increasing L-carnitine intake, the endogenous carnitine pool may not significantly increase. Therefore,
the FEEDAP Panel considers the use of L-carnitine and L-carnitine L-tartrate as additives in animal
nutrition to be safe for the consumer.
L-Carnitine and L-carnitine L-tartrate are not irritant to skin and eyes, nor are they skin sensitisers. L-
Carnitine and L-carnitine L-tartrate show limited dust formation. As inhalation toxicity studies are not
available, adverse effects in the respiratory tract cannot be fully excluded.
The use of L-carnitine and L-carnitine L-tartrate in animal nutrition is not expected to pose a risk to the
environment.
L-Carnitine and L-carnitine L-tartrate are regarded as effective sources of L-carnitine in all animal
species. Nutritional requirement data for the different animal species and categories have not been
established.
RECOMMENDATIONS
The FEEDAP Panel proposes to use the specifications for L-carnitine according to PhEur (MG 1339)
considering purity, substance-related impurities and other impurities (sulphated ash and heavy metals).
DOCUMENTATION PROVIDED TO EFSA
1. L-Carnitine and L-carnitine L-tartrate as a feed additive for all animal species. October 2010.
Submitted by Lonza Benelux BV.
2. L-Carnitine and L-carnitine L-tartrate as a feed additive for all animal species. Supplementary
information. October 2011. Submitted by Lonza Benelux BV.
3. L-Carnitine and L-carnitine L-tartrate as a feed additive for all animal species. Supplementary
information. February 2012. Submitted by Lonza Benelux BV.
4. Evaluation report of the European Union Reference Laboratory for Feed Additives on the
methods(s) of analysis for L-carnitine and L-carnitine L-tartrate.
5. Comments from Member States received through the ScienceNet.
REFERENCES
AWT (Arbeitsgemeinschaft fr Wirkstoffe in der Tierernhrung e.V.), 2002. Vitamins in Animal
Nutrition. ISBN 3-86037-167-3.
Bremer J, 1983. Carnitine metabolism and functions. Physiology Review, 63, 14201480.
L-Carnitine and L-carnitine L-tartrate for all animal species

EFSA Journal 2012;10(5):2676 15
Birkenfeld C, Kluge H and Eder K, 2006. L-Carnitine supplementation of sows during pregnancy
improves the suckling behaviour of their offspring. British Journal of Nutrition, 96, 334342.
Borghijs HK and DeWilde RO, 1992. The influence of two different dosages of L-carnitine on some
blood parameters during exercise in trained pigeons. Journal of Veterinary Nutrition, 1, 3135.
Brooks DE and McIntosh JE, 1975. Turnover of carnitine by rat tissues. Biochemical Journal, 148,
439445.
Burtle GJ, 1990. L-Carnitine enhanced fish growth. Presentation at Carnitine seminar, Lonza, Basel.
Burtle GJ, 1993. Effects of dietary L-carnitine supplements on growth and muscle lipid of fingerling
channel catfish. Report to Lonza, Basel.
Carroll JE, Brooke MH, Shumate JB and Janes NJ, 1981. Carnitine intake and excretion in
neuromuscular diseases. American Journal of Clinical Nutrition, 34, 26932698.
Demarquoy J, Georges B, Rigault C, Royer M-C, Clairet A, Soty M, Lekounoungou S and Le Borgne
F, 2004. Radioisotopic determination of L-carnitine content in foods commonly eaten in Western
countries. Food Chemistry, 86, 137142.
Dubelaar ML, Lucas CM and Hlsmann WC, 1991a. The effect of L-carnitine on force development
of the latissimus dorsi muscle in dogs. Journal of Cardiac Surgery, 6, 270275.
Dubelaar ML, Lucas CM and Hlsmann WC, 1991b. Acute effect of L-carnitine on skeletal muscle
force tests in dogs. American Journal of Physiology 260, E189193.
EFSA (European Food Safety Authority), 2003. Opinion of the Scientific Panel on Food Additives,
Flavourings, Processing Aids and Materials in Contact with Food (AFC) related to L-carnitine-L-
tartrate for use in foods for particular nutritional purposes. EFSA Journal 19, 113.
EFSA Panel on Dietetic Products, Nutrition and Allergies (NDA), 2011. Scientific opinion on the
substantiation of health claims related to L-carnitine and faster recovery from muscle fatigue after
exercise (ID 738, 1492, 1493), skeletal muscle tissue repair (ID 738, 1492, 1493), increase in
endurance capacity (ID 4305, 4684), maintenance of normal blood LDL-cholesterol concentrations
(ID 1494, 4684), contribution to normal spermatogenesis (ID 1822), energy metabolism (ID
1821), and increasing L-carnitine concentrations and/or decreasing free fatty acids in blood during
pregnancy (ID 1495) pursuant to Article 13(1) of Regulation (EC) No 1924/2006. EFSA Journal
9(6): 2212, 24pp.
European Pharmacopeia (PhEur), 2010. Levocarnitine, Monograph (MG) 1339, 7th edition.
Strasbourg, France: Council of Europe (COE) European Directorate for the Quality of Medicines.
Falaschini A and Trombetta MF, 1993. Studio sulla possibilit di impiego della L-carnitina nel cavallo
da trotto. Research report to LONZA, University of Ancona.
Fischer M, Varady J, Hirche F, Kluge H and Eder K, 2009. Supplementation of L-carnitine in pigs:
absorption of carnitine and effect on plasma and tissue carnitine concentrations. Archives of
Animal Nutrition, 63, 115.
Foster CV, Harris RC and Snow DH, 1988. The effect of oral L-carnitine supplementation on the
muscle and plasma concentrations in the Thoroughbred horse. Comparative Biochemistry and
Physiology. Part A. Comparative Physiology, 91, 827835.
Gevaert D, Vervaet J and De Wilde R, 1991. Effect of training and L-carnitine on long-term exercise
in pigeon. Kurzfassung, 44. Tagung der Gesellschaft fr Ernhrungsphysiologie der Haustiere, 89.
Ghler K, 2002. A study on the carnitine metabolism of early weaned piglets. Dissertation vet. med.,
Faculty of Veterinary Medicine, University of Leipzig, Germany, 94pp.
Gtz C, 1989. Bestimmung von L-Carnitin und seiner Ester in Lebensmitteln. Hamburg, University,
Fachbereich Chemie, Dissertation.
L-Carnitine and L-carnitine L-tartrate for all animal species

EFSA Journal 2012;10(5):2676 16
Gudjonsson H, Li BU, Shug AL and Olsen WA, 1985. In vivo studies of intestinal carnitine absorption
in rats. Gastroenterology, 88, 18801887.
Gnther A, 1989. L-Carnitine trial with fattening pigs. University of Gttingen, Report to Lonza,
Basel.
Hamilton JW, Li BU, Shug AL and Olsen WA, 1986. Carnitine transport in human intestinal biopsy
specimens. Demonstration of an active transport system. Gastroenterology, 91, 1016.
Harper P, Elwin CE and Cederblad G, 1988. Pharmacokinetics of intravenous and oral bolus doses of
L-carnitine in healthy subjects. European Journal of Clinical Pharmacology, 35, 555562.
Harris RC, Forster CVL and Snow DH, 1995. Plasma carnitine concentration and uptake into muscle
following oral and intravenous administration. Equine Veterinary Journal, 18, 382-387.
Heo K, Odle J, Han IK, Cho W, Seo S, van Heugten E and Pilkington DH, 2000. Dietary L-carnitine
improves nitrogen utilization in growing pigs fed low energy, fat-containing diets. Journal of
Nutrition, 130, 18091814.
Jger U, 2000. A study on the carnitine status of baby quail (Coturnix coturnix japonica). Dissertation
vet. med., Faculty of Veterinary Medicine, University of Leipzig, Germany, 94pp.
Ji H, Bradley TM and Tremblay GC, 1996. Atlantic salmon (Salmo salar) fed L-carnitine exhibit
altered intermediary metabolism and reduced tissue lipid, but no change in growth rate. Journal of
Nutrition, 126, 19371950.
Kliemant A, 2000. Studies on the carnitine metabolism of Japanese quail (Coturnix coturnix japonica).
Dissertation vet. med., Faculty of Veterinary Medicine, University of Leipzig, Germany, 87pp.
Knttel-Gustavsen S and Harmeyer J, 2007. The determination of L-carnitine in several food samples.
Food Chemistry, 105, 793-804.
Li BU, Bummer PM, Hamilton JW, Gudjonsson H, Zografi G and Olsen WA, 1990. Uptake of L-
carnitine by rat jejunal brush border microvillous membrane vesicles. Evidence of passive
diffusion. Digestive Diseases and Sciences, 35, 333339.
Lindstedt G and Lindstedt S, 1961. On the biosynthesis and degradation of carnitine. Biochemical and
Biophysical Research Communications, 6, 319323.
Litz H, 1993. Examination of the overall effect of a L-carnitine addition to piglet starter diets and to
diets for rainbow trout (Oncorhynchus mykiss). Dissertation med. vet., University of Munich,
Munich, Germany, 61pp.
Marciani P, Lindi C, Marzo A, Martelli EA, Cardace G and Esposito G, 1991. L-Carnitine and
carnitine ester transport in the rat small intestine. Pharmacological Research, 23, 157162.
McDowell LR, 2000. Vitamins in Animal and Human Nutrition, 2nd edition. Ames, IA: Iowa State
University Press, 645 pp. ISBN 0-8138-2630-6.
Mehlman MA, Kader MM and Therriault DG, 1969. Metabolism, turnover time, half life, body pool
of carnitine-14C in normal, alloxan diabetic and insulin treated rats. Life Sciences, 8, 465472.
Mitchell ME, 1978. Carnitine metabolism in human subjects. I. Normal metabolism. American Journal
of Clinical Nutrition, 31, 293306.
Musser RE, Goodband RD, Tokach MD, Owen KQ, Nelssen JL, Blum SA, Dritz SS and Civis CA,
1999a. Effects of L-carnitine fed during gestation and lactation on sow and litter performance.
Journal of Animal Science, 77, 32893295.
Musser RE, Goodband RD, Tokach MD, Owen KQ, Nelssen JL, Blum SA, Campbell RG, Smits R,
Dritz SS and Civis CA, 1999b. Effects of L-carnitine fed during lactation on sow and litter
performance. Journal of Animal Science, 77, 32963303.
Newton GL and Haydon KD, 1986. Carnitine in nursery pig diets. University of Georgia Swine
Report, College of Agriculture, Georgia. Special Publ., 41, 3639.
L-Carnitine and L-carnitine L-tartrate for all animal species

EFSA Journal 2012;10(5):2676 17
Newton, GL and Haydon KD, 1988. Carnitine in nursery pig diets. Journal of Animal Science,
66(Suppl. 1), abstr. 40.
Newton, GL and Haydon KD, 1989. Carnitine supplementation for finishing pigs. Journal of Animal
Science, 67(Suppl. 1), abstr. 267.
Owen KQ, Nelssen JL, Goodband RD, Tokach MD, Blum SA, Dritz SS and Musser R, 1994. The
effect of dietary L-carnitine on growth performance and tissue accretion rates in the early weaned
pig. Journal of Animal Science, 72(Suppl. 2), abstr. 70.
Owen KQ, Nelssen JL, Goodband RD, Weeden TL and Blum SA, 1996. Effect of L-carnitine and
soybean oil on growth performance and body composition of early-weaned pigs. Journal of Animal
Science, 74, 16121619.
Owen KQ, Nelssen JL, Goodband RD, Tokach MD and Friesen KG, 2001a. Effect of dietary L-
carnitine on growth performance and body composition in nursery and growing-finishing pigs.
Journal of Animal Science, 79, 15091515.
Owen KQ, Jit H, Maxwell CV, Nelssen JL, Goodband RD, Tokach MD, Tremblay GC and Koo SI,
2001b. Dietary L-carnitine suppresses mitochondrial branched-chain keto acid dehydrogenase
activity and enhances protein accretion and carcass characteristics of swine. Journal of Animal
Science 79, 31043112.
Pelletier B, 1992. L-Carnitine or vitamin Bt of interest in the dog. Action Vterinaire, 1210, 1921.
Ramanau A, Kluge H, Spilke J and Eder K, 2004. Supplementation of sows with L-carnitine during
pregnancy and lactation improves growth of the piglets during the suckling period through
increased milk production. Journal of Nutrition, 134, 8692.
Ramanau A, Kluge H and Eder K, 2005. Effects of L-carnitine supplementation on milk production,
litter gains and back-fat thickness in sows with a low energy and protein intake during lactation.
British Journal of Nutrition, 93, 717721.
Rebouche CJ and Engel AG, 1984. Kinetic compartmental analysis of carnitine in the human carnitine
deficiency syndromes. Evidences for alterations in tissue carnitine transport. Journal of Clinical
Investigation, 73, 857867.
Rebouche CJ and Chenard CA, 1991. Metabolic fate of dietary carnitine in human adults:
identification and quantification of urinary and fecal metabolites. Journal of Nutrition, 121, 539
46.
Rebouche CJ, 2004. Kinetics, pharmacokinetics, and regulation of L-carnitine and acetylL-carnitine
metabolism. Annals of the New York Academy of Sciences, 1033, 3041.
Rincker MJ, Carter SD, Real DE, Nelssen JL, Tokach MD, Goodband RD, Dritz SS, Senne BW, Fent
RW, Pettey LA and Owen KQ, 2003. Effects of increasing dietary L-carnitine on growth
performance of weanling pigs. Journal of Animal Science, 81, 22592269.
Santulli A and DAmelio V, 1986. Effects of supplemental dietary carnitine on growth and lipid
metabolism of hatchery-reared sea bass (Dicentrarchus labrax L.). Aquaculture, 59, 177186.
Santulli A, Modica A, Curatolo A and DAmelio V, 1988. Carnitine administration to sea bass
(Dicentrarchus labrax L.) during feeding on a fat diet: modification of plasma lipid levels and
lipoprotein levels. Aquaculture, 68, 345351.
Santulli A, Puccia E and DAmelio V, 1990. Preliminary study on the effect of shortterm carnitine
treatment on nucleic acids and protein metabolism in sea bass (Dicentrarchus labrax L.) fry.
Aquaculture, 87, 8599.
Schnitger A, 2003. Effect of oral carnitine supplementation on plasma carnitine concentration in the
horse. Dissertation vet. med., Faculty of Veterinary Medicine, University of Leipzig, Germany, 71
pp.
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EFSA Journal 2012;10(5):2676 18
Schuhmacher A and Gropp J, 1998. Carnitine a vitamin for rainbow trout? Journal of Applied
Ichthyology, 14, 8790.
Schuhmacher A, Eissner C and Gropp J, 1993. Carnitin bei Fischen, Ferkeln und Wachteln. In:
Flachowsky G, Schubert R (eds), Vitamine und weitere Zusatzstoffe. Niederkleen, Germany:
Wissenschaftlicher Fachverlag Dr. Fleck, pp. 407-412
Shaw RD, Li BU, Hamilton JW, Shug AL and Olsen WA, 1983. Carnitine transport in rat small
intestine. American Journal of Physiology, 245, G376G381.
Torrelle E, VanDer Sluiszen A and Verreth J, 1991. The effect of dietary L-carnitine on the growth
performance of the African catfish (Clarias gariepinus) in relation to dietary lipid. British Journal
of Nutrition, 69, 28999.
Yue KT and Fritz IB, 1962. Fate of tritium-labeled carnitine administered to dogs and rats. American
Journal of Physiology, 202, 122-128.
L-Carnitine and L-carnitine L-tartrate for all animal species

EFSA Journal 2012;10(5):2676 19
APPENDICES
APPENDIX A
Executive Summary of the Evaluation Report of the European Union Reference Laboratory for
Feed Additives on the Method(s) of Analysis for L-Carnitine and L-Carnitine L-Tartrate
36

In the current application authorisation is sought under article 4(1) and 10(2) for L-Carnitine
1,2
and L-
Carnitine L-Tartrate (LCLT)
2
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 Annex I of Regulation (EC) No 1831/2003. Authorisation is
sought for the use of the two feed additives for all animal species and categories. According to the
Applicants, L-Carnitine and LCLT have a minimum purity of 97 %. Both feed additives are intended
to be used in premixtures or added directly into the feedingstuffs and water. None of the Applicants
proposed any minimum or maximum concentration of L-Carnitine or LCLT in feedingstuffs or water,
as set in the previous regulations.
For the determination of L-Carnitine in the feed additive, Applicant
1
submitted the US Pharmacopoeia
method. The EURL identified instead the European Pharmacopoeia method. Even though no
performance characteristics are provided, the EURL recommends for official control the European
Pharmacopoeia method (Ph. Eur. 6
th
Edition, monographs 1339), for the identification and
quantification of L-Carnitine in the feed additive.
For the determination of L-Carnitine in premixtures, feedingstuffs and water Applicant
1
submitted an
enzymatic method, specific for the L-Carnitine isomer. This single-laboratory validated method was
further verified by a second independent laboratory. The relative precisions (i.e. relative standard
deviations of repeatability (RSD
r
) and relative standard deviations of intermediate precision (RSD
ip
)
ranging from 4.5 to 5.1 % were recalculated by the EURL using the experimental data provided by the
Applicant
1
. Furthermore, the Applicant
1
reported a recovery rate (R
Rec
) ranging from 94 to 102 % and
a limit of detection (LOD) of 15 mg L-Carnitine /kg feedingstuffs, which is well below the usual
concentrations of L-Carnitine in feedingstuffs or water.The experiments were carried out at 5 g L-
Carnitine /kg premixtures and 100-165 mg L-Carnitine /kg feedingstuffs or water.
For the determination of L-Carnitine in premixtures, Applicant
2
proposed an ion chromatography
method with electrical conductivity detection (IC-ECD), which does not distinguish between two
enantiomers L- and D-Carnitine. This single-laboratory validated method was further verified by a
second independent laboratory. The relative precisions ranging from 4.2 to 4.6 % were recalculated by
the EURL using the experimental data provided by the Applicant
2
. Applicant
2
reported R
Rec
ranging
from 92 to 100 %. The experiments were carried out at 8-10 g L-Carnitine /kg premixtures.
For the determination of L-Carnitine in feedingstuffs, Applicant
2
proposed a Reversed-Phase High
Performance Liquid Chromatography method using a fluorimetric detector (RP-HPLC), which does
not distinguish between two enantiomers L- and D-Carnitine. The relative precisions ranging from 6.9
to 12.4 % and LOD of 6.3 mg L-Carnitine /kg feedingstuffs were recalculated by the EURL using the
experimental data provided by the Applicant
2
. Applicant
2
reported R
Rec
ranging from 88 to 115 %. The
experiments were carried out at 40-60 mg L-Carnitine /kg feedingstuffs.
For the determination of L-Carnitine in water, Applicant
2
proposed a potentiometric titration. RSD
r
of
0.1 % was reported by Applicant
2
.
Based on the above considerations and the performance characteristics presented, the EURL
recommends for official control the following single-laboratory validated and further verified methods

36
The full report is available on the EURL website. http://irmm.jrc.ec.europa.eu/SiteCollectionDocuments/FinRep-2010-
0144+0225.pdf
a
FAD-2010-0225;
b
FAD-2010-0144.
L-Carnitine and L-carnitine L-tartrate for all animal species

EFSA Journal 2012;10(5):2676 20
based on the spectrophotometric method after enzymatic reaction with carnitine-acetyl-transferase
enzymatic reaction for the determination of L-Carnitine in premixtures, feedingstuffs and water,
together with
ion chromatography method with electrical conductivity detection (IC-ECD) for the
determination of L-Carnitine in premixtures;
Reversed-Phase High Performance Liquid Chromatography (RP-HPLC) using a fluorimetric
detector for the determination of L-Carnitine in feedingstuffs;
potentiometric titration with hydrochloric acid for the determination of L-Carnitine in the
water.
For the determination of L-Carnitine L-Tartrate (LCLT) in the feed additive, Applicant
2
submitted a
potentiometric back-titration. The method was single-laboratory validated and further verified. RSD
r

ranging from 0.72 to 1.19 % and RSD
ip
of 1.19 were recalculated by the EURL using the experimental
data provided by the Applicant
2
. R
Rec
ranging from 99.8 to 101 % were reported by the Applicant
2
.
Based on the performance characteristics presented, the EURL recommends for official control the
single-laboratory validated and further verified potentiometric back-titration method to determine L-
Carnitine L-Tartrate (LCLT) in the feed additive.
L-Carnitine is released from the LCLT salt during sample treatment. Therefore, the EURL considers
that all the analytical methods recommended for the determination of L-Carnitine in the matrices
investigated are suitable for official control to determine LCLT (expressed as L-Carnitine) in
premixtures, feedingstuffs and 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.

L-Carnitine and L-carnitine L-tartrate for all animal species

EFSA Journal 2012;10(5):2676 21
APPENDIX B
L-Carnitine its physiological role, endogenous synthesis and metabolism
L-Carnitine, a quaternary amine, has a key role in the metabolism of fatty acids (energy) as a
substrate for the reversible acetylation of coenzyme A (Friedmann and Fraenkel, 1955) and as a
carrier for the transport of long-chain fatty acids from cytosol across the inner mitochondrial
membrane (Fritz and Marquis, 1965). Furthermore, it is suggested that L-carnitine improves the
oxygen supply and utilisation of the skeletal and heart muscle (Liedtke et al., 1982; Shug, 1987;
Pierpont, 1992; Broderick et al., 1995; Neu, 1995). Accordingly, carnitine might also be important in
the case of muscle exercise, particularly under anaerobic conditions (Cerretelli and Marconi, 1988).
L-Carnitine is synthesised endogenously in five enzyme-catalysed steps from its precursor lysine,
which provides the carbon backbone, and with methionine as methyl group donor (e.g. Bremer, 1961,
1962; Wolf and Berger, 1961; Tanphaichitr et al., 1973; Cox and Hoppel, 1973a,b, 1974; Horne and
Broquist, 1973). The principles of the synthesis and metabolism of L-carnitine are given in Figure B1.


Figure B1: L -Carnitine synthesis and schematic metabolism and function of carnitine

L-Carnitine and L-carnitine L-tartrate for all animal species

EFSA Journal 2012;10(5):2676 22
In juvenile organisms this synthesis is probably insufficient to meet endogenous carnitine
requirements, possibly due to the low activity of -butyrobetaine-hydroxylase (Rebouche and Engel,
1980; Hahn, 1981) that occurs mainly in the liver and catalyses the last step of carnitine formation.
The activity of the enzyme starts to increase about 48 days after birth, and the highest concentrations
are reached in adults (Hahn, 1981). Based on these observations carnitine is sometimes considered to
be conditionally essential. Dietary supplementation with L-carnitine is therefore expected to improve
intermediary fatty acid and energy utilisation and therefore weight gain and feed efficiency in young
animals, particularly when diets marginally deficient in lysine and/or methionine are fed. However, up
until now it has not been possible to define the requirement for L-carnitine.
After ingestion L-carnitine is absorbed in the small intestine via a sodium-dependent transport system
(low carnitine concentration) or at pharmacological doses of carnitine via passive diffusion as free
carnitine and short-chain carnitine esters (Shaw et al., 1983; Gudjonsson et al., 1985; Hamilton et al.,
1986; Li et al., 1990; Marciani et al., 1993). The bioavailability of dietary L-carnitine is about 54
87 % in humans and is dependent on the dietary L-carnitine content (Rebouche and Chenard, 1991;
Rebouche, 2004). The major portion of all carnitine in the body is found in the skeletal muscle (see,
for example, Mitchell, 1978). Studies with labelled L-carnitine suggest that carnitine is usually largely
not metabolised (e.g. Lindstedt and Lindstedt, 1961; Yue and Fritz, 1962; Brooks and MacIntosh,
1975; Bremer, 1983); however, carnitine might be converted, for example to -methyl choline, during
stress and disease or depending on the dietary or physiological conditions (Mehlman et al., 1969;
Mitchell, 1978). Carnitine is excreted mainly via the kidney with a highly efficient tubular
reabsorption of about 9099 % at normal dietary intakes and normal plasma and tissue concentrations
(Harper et al., 1988; Rebouche, 2004); only 2 % of the ingested L-carnitine is voided in the faeces
(Carroll et al., 1981).
REFERENCES
Bremer J, 1961. Biosynthesis of carnitine in vivo. Biochimica et Biophysica Acta, 48, 622624.
Bremer J, 1962. Carnitine precursors in the rat. Biochimica et Biophysica Acta, 57, 327335.
Bremer J, 1983. Carnitine metabolism and functions. Physiology Review, 63, 14201480.
Broderick TL, Christos SC, Wolf BA, DiDiomenico D, Shug AL and Paulson DJ, 1995. Fatty acid
oxidation and cardiac function in the sodium pivalate model of secondary carnitine deficiency.
Metabolism, 44, 499505.
Brooks DE and McIntosh JE, 1975. Turnover of carnitine by rat tissues. Biochemical Journal, 148,
439445.
Carroll JE, Brooke MH, Shumate JB and Janes NJ, 1981. Carnitine intake and excretion in
neuromuscular diseases. American Journal of Clinical Nutrition, 34, 26932698.
Cerretelli P and Marconi C, 1988. L-Carnitine supplementation in humans. The effects on physical
performance. Lonza Documentation, Lonza, Basel.
Cox RA and Hoppel CL, 1973a. Biosynthesis of carnitine and 4-N-trimethylaminobutyrate from
lysine. Biochemical Journal, 136, 10751082.
Cox RA and Hoppel CL, 1973b. Biosynthesis of carnitine and 4-N-trimethylaminobutyrate from 6-N-
trimethyl-lysine. Biochemical Journal, 136, 10831090.
Cox RA and Hoppel CL, 1974. Carnitine and trimethylaminobutyrate synthesis in rat tissues.
Biochemical Journal, 142, 699701.
Friedman S and Fraenkel G, 1955. Reversible enzymatic acetylation of carnitine. Archives of
Biochemistry and Biophysics, 59, 491501.
Fritz IB and Marquis NR, 1965. The role of acylcarnitine esters and carnitine palmityltransferase in
the transport of fatty acyl groups across mitochondrial membranes. Biochemistry, 54, 12261233.
L-Carnitine and L-carnitine L-tartrate for all animal species

EFSA Journal 2012;10(5):2676 23
Gudjonsson H, Li BU, Shug AL and Olsen WA, 1985. In vivo studies of intestinal carnitine absorption
in rats. Gastroenterology, 88, 18801887.
Hahn P, 1981. The development of carnitine synthesis from -butyrobetaine in the rat. Life Sciences,
29, 10571060.
Hamilton JW, Li BU, Shug AL and Olsen WA, 1986. Carnitine transport in human intestinal biopsy
specimens. Demonstration of an active transport system. Gastroenterology, 91, 1016.
Harper P, Elwin CE and Cederblad G, 1988. Pharmacokinetics of intravenous and oral bolus doses of
L-carnitine in healthy subjects. European Journal of Clinical Pharmacology, 35, 555562.
Horne DW and Broquist HP, 1973. Role of lysine and -N-trimethyllysine in carnitine bio-synthesis. I.
Studies in Neurospora crassa. Journal of Biological Chemistry, 248, 21702175.
Li BU, Bummer PM, Hamilton JW, Gudjonsson H, Zografi G and Olsen WA, 1990. Uptake of L-
carnitine by rat jejunal brush border microvillous membrane vesicles. Evidence of passive
diffusion. Digestive Diseases and Sciences, 35, 333339.
Liedtke AJ, Nellis SH, Whitesell LF and Mahar SQ, 1982. Metabolic and mechanical effects using L-
and D-carnitine in working swine hearts. American Journal of Physiology, 243, H691H697.
Lindstedt G and Lindstedt S, 1961. On the biosynthesis and degradation of carnitine. Biochemical and
Biophysical Research Communications, 6, 31923.
Marciani P, Lindi C, Marzo A, Martelli EA, Cardace G and Esposito G, 1991. L-Carnitine and
carnitine ester transport in the rat small intestine. Pharmacology Research, 23, 15762.
Mehlman MA, Kader MM and Therriault DG, 1969. Metabolism, turnover time, half life, body pool
of carnitine-14C in normal, alloxan diabetic and insulin treated rats. Life Sciences, 8, 465472.
Mitchell ME, 1978. Carnitine metabolism in human subjects. I. Normal metabolism. American Journal
of Clinical Nutrition, 31, 293306.
Neu H, 1995. Carnitin: Chemie, Funktion und klinische Bedeutung bei Herzerkrankungen
(Kardiomyopathien) des Hundes eine Literaturbersicht. Kleintierpraxis, 3, 197220.
Pierpont ME, 1992. Carnitine and myocardial function. In: Carter AL (ed.), Current concepts in
carnitine research. Boca Raton, FL: CRS Press, pp. 192212.
Rebouche CJ and Engel AG, 1980. Tissue distribution of carnitine biosynthetic enzymes in man.
Biochimica Biophysica Acta, 630, 2229.
Rebouche CJ and Chenard CA, 1991. Metabolic fate of dietary carnitine in human adults:
identification and quantification of urinary and fecal metabolites. Journal of Nutrition, 121, 539
546.
Rebouche CJ, 2004. Kinetics, pharmacokinetics, and regulation of L-carnitine and acetyl-L-carnitine
metabolism. Annals of the New York Academy of Sciences, 1033, 3041.
Shaw RD, Li BU, Hamilton JW, Shug AL and Olsen WA, 1983. Carnitine transport in rat small
intestine. American Journal of Physiology, 245, G376381.
Shug AL, 1987. Protection from adriamycin-induced cardiomyopathy in rats. Zeitschrift fr
Kardiologie, 76(Suppl. 5), 4652.
Tanphaichitr V, Broquist HP, 1973. Role of lysine and -N-trimethyllysine in carnitine biosynthesis.
II. Studies in the rat. Journal of Biological Chemistry, 248, 21762181.
Wolf G and Berger CR, 1961. Studies on the biosynthesis and turnover of carnitine. Archives of
Biochemistry and Biophysics, 92, 360365.
Yue KT and Fritz IB, 1962. Fate of tritium-labeled carnitine administered to dogs and rats. American
Journal of Physiology, 202, 122128.

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