#221 
 
Guidance for Industry 
 
Recommendations for Preparation and Submission of 
Animal Food Additive Petitions 
 
Submit comments on this guidance at any time.  Submit  written comments to the 
Division  of Dockets Management (HFA-305), Food and Drug Administration,  5630 
Fishers Lane, Room 1061, Rockville,  MD 20852.  Submit electronic comments on the 
guidance at http://www.regulations.gov.   All written comments should be identified  with 
the Docket No. FDA-2013-D-0928. 
 
For further information  regarding this document, contact Mika Alewynse, Center for 
Veterinary Medicine, Division  of Animal Feeds (HFV-229), Food and Drug 
Administration,  7519 Standish Place, Rockville,  MD 20855, 240-402-5843;  email: 
Mika.Alewynse@fda.hhs.gov. 
 
Additional  copies of this guidance document may be requested from the Policy and 
Regulations Staff (HFV-6), Center for Veterinary Medicine, Food and Drug 
Administration,  7519 Standish Place, Rockville,  MD 20855, and may be viewed on the 
Internet at either http://www.fda.gov/AnimalVeterinary/default.htm  or 
http://www.regulations.gov. 
 
U.S. Department of Health and Human Services 
Food and Drug Administration 
Center for Veterinary Medicine 
June 2015 
 
   
  Contains Nonbinding Recommendations 
Table of Contents 
 
I. INTRODUCTION ...................................................................................................3 
II. LEGAL AUTHORITY ..........................................................................................3 
III. GENERAL CONSIDERATIONS AND THE FAP PROCESS ............................4 
A. Prior to Submitting a Petition .....................................................................4 
B. Format and Information .............................................................................5 
C. Identity and Composition ...........................................................................6 
D. Intended Use and Use Level, and Labeling ...............................................10 
E. Data Establishing the Intended Effect.......................................................11 
F. Analytical Methods....................................................................................12 
G. Safety Evaluation ......................................................................................12 
H. Proposed Tolerances for the Food Additive .............................................15 
I. Proposed Regulation ..................................................................................15 
J. Environmental Assessment ........................................................................15 
IV. OTHER INFORMATION ..................................................................................17 
APPENDIX 1 ............................................................................................................18 
 
   
2  Contains Nonbinding Recommendations 
Guidance for Industry 
 
Recommendations  for Preparation  and Submission  of 
Animal  Food Additive Petitions 
 
This guidance represents the current thinking of the Food and Drug Administration (FDA 
or Agency) on this topic.  It does not establish any rights for any person and is not 
binding on FDA or the public.  You can use an alternative approach if it satisfies the 
requirements of the applicable statutes and regulations.  To discuss an alternative 
approach, contact the FDA staff responsible for this guidance as listed on the title page. 
 
I. INTRODUCTION 
 
This guidance describes the types of information  that the Food and Drug 
Administrations  (FDAs) Center for Veterinary Medicine (CVM) recommends for 
inclusion  in food additive petitions  (FAPs) for food additives used in animal food.  This 
guidance only applies to food additives for use in animal food.  It is intended to help the 
petitioner (you) submit FAP information  in a consistent and appropriate manner. 
 
This guidance is intended to be used in conjunction  with other CVM Guidance for 
Industry (GFI) documents that are cited throughout this document. Some of the 
referenced GFI documents pertain to animal drug products.  These guidance documents 
are referenced because they provide information  that is relevant to FDAs evaluation of 
the safety of food additives used in animal food.  For example, FDA evaluates the human 
food safety of tissue residues present in edible tissues in a similar  manner for new animal 
drugs intended for use in food-producing  animals and for food additives used in foods for 
food-producing  animals. 
 
FDAs guidance documents, including  this guidance, do not establish legally  enforceable 
responsibilities.   Instead, guidances describe FDAs current thinking  on various topics 
and should be viewed only as recommendations,  unless specific regulatory or statutory 
requirements are cited.  The use of the word should in FDA guidances means that 
something is suggested or recommended, but not required. 
 
II. LEGAL AUTHORITY 
 
FDA regulates foods and substances added to food under the Federal Food, Drug, and 
Cosmetic Act (FD&C Act).  Many substances added to foods are food additives.  Section 
201(s) of the FD&C Act (21 U.S.C.  321(s)) defines a food additive as any substance 
the intended use of which results or may reasonably be expected to result, directly or 
indirectly,  in its becoming a component of or otherwise affecting the characteristics of 
any food. Section 201(s) excludes from this definition  any substance that is generally 
3  Contains Nonbinding Recommendations 
recognized as safe (GRAS) among qualified  experts under the conditions  of its intended 
use, or that qualifies for any of the other exemptions from the food additive definition 
(e.g., new animal drug, color additive,  etc.).1 
 
Food additives require premarket approval based on data demonstrating safety.  FDA 
issues food additive regulations specifying  the conditions  under which an additive has 
been demonstrated to be safe and, therefore, may be lawfully  used.  (See Section 
409(a)(2) of the FD&C Act (21 U.S.C.  348(a)(2)).  A food that contains an unapproved 
food additive is deemed to be unsafe under section 409(a) of the FD&C Act and 
adulterated under section 402(a)(2)(C)(i) of the FD&C Act.  Adulterated foods cannot be 
legally  imported or marketed in the United States.  Therefore, to use a food additive in 
animal food, FDA must issue a food additive regulation prescribing  the conditions  under 
which the additive  may be safely used.  Section 409(b) through (g) of the FD&C Act 
provides for a petition  process to establish that a use of a food additive is safe. 
 
Pursuant to Section 409(j) of the FD&C Act (21 U.S.C.  348(j)), a food additive,  or food 
containing  a food additive,  intended for investigational  use may be exempt from the 
requirements of section 409 of the FD&C Act under the conditions  set forth in Title 21 of 
the Code of Federal Regulations  section 570.17.2 
 
Under section 201(s) of the FD&C Act, the use of a food substance may be GRAS either 
through scientific procedures or, for a substance used in food before 1958, through either 
scientific procedures or experience based on common use in food.  General recognition  of 
safety through scientific procedures requires the same quantity and quality  of scientific 
evidence as is needed for approval of an FAP (21 CFR 570.30(b)).  The implementing 
regulations for eligibility  for GRAS classification  and affirmation of GRAS status for 
animal food are found at 21 CFR 570.30 and 21 CFR 570.35. 
 
III. GENERAL CONSIDERATIONS AND THE FAP PROCESS 
 
A. Prior to Submitting a Petition 
 
Prior to submitting  an FAP, CVM recommends that you verify that the substance you 
wish to use in animal food is not already approved as a food additive  for your intended 
use.  CVM publishes  food additive regulations at 21 CFR parts 573 and 579.  
                                              
1 Under section 201(s) of the FD&C Act, the following types of substances are excluded from the 
food additive definition:  (1) pesticide chemical residues in or on a raw agricultural commodity or 
processed food; (2) pesticide chemicals; (3) color additives; (4) substances used in accordance 
with a prior sanction (i.e., a sanction or approval granted prior to the enactment of the Food 
Additives Amendment of 1958 under the FD&C Act, the Poultry Products Inspection Act, or the 
Meat Inspection Act; (5) new animal drugs; and (6) dietary ingredients in or intended for use in a 
dietary supplement. 
2 The Code of Federal Regulations is available on the internet at http://www.ecfr.gov/cgi-
bin/ECFR?page=browse. 
4  Contains Nonbinding Recommendations 
Furthermore, some substances that are GRAS for an intended use are listed in 21 CFR 
parts 582 and 584. 
 
To be acceptable for a particular application,  a food additive must meet the requirements 
(e.g., identity,  specifications,  and use limitations)  set forth in 21 CFR part 571. 
 
If you determine that you need to submit an FAP, CVM encourages pre-petition 
consultations,  which could include requests for meetings or teleconference calls, to 
facilitate the development  of information  needed for an FAP, particularly  for those that 
do not have previous experience preparing an FAP for animal food purposes.  CVM also 
recommends that you initiate contact with CVM early in the process.  A meeting or 
telephone conference with CVM staff to discuss a research plan for the proposed food 
additive is often useful. Once a research plan is developed,  CVM strongly suggests that 
you submit protocols for CVM's review before initiating  the studies. 
 
Due to the wide variety of food additives and their intended uses, FDA cannot offer, in 
this guidance, detailed recommendations  for some aspects of the FAP process.  The FAP 
process is an iterative process and FDA offers detailed guidance to petitioners  during the 
process.  Such guidance depends on the type of food additive and its intended use.  If 
your food additive  involves the use of a specific technology (e.g., nanotechnology),  CVM 
recommends you consult the FDA website for additional  guidances pertaining to the 
application  of that technology to products that fall under FDAs regulatory purview.3 
 
If you need assistance determining  the status of an animal food ingredient  or preparing an 
FAP you should contact the Division  of Animal Feeds in CVM in the following  ways: 
 
Mail: 
Center for Veterinary Medicine, 
Division  of Animal Feeds, 
7519 Standish Place, HFV-220, 
Rockville,  MD 20855 
 
Phone: 240-402-7077 
Fax:  240-453-6882 
Email:  Mika.Alewynse@fda.hhs.gov 
 
B. Format and Information 
 
Section 409(b)(2) of the FD&C Act (21 U.S.C.  348(b)(2)) and FDAs implementing 
regulations at 21 CFR part 571 describe the FAP process and the information  that must 
be submitted to FDA as part of an FAP.  FDA reviews the submitted information  and 
assesses whether, as a whole, the data and information  demonstrate the safety of the food 
additive for an intended use as requested in the petition. 
                                              
3 See guidances at http://www.fda.gov/RegulatoryInformation/Guidances/default.htm. 
5  Contains Nonbinding Recommendations 
The data and information  in an FAP that are available for public disclosure are described 
in 21 CFR 571.1(h)(1).  The data and information  in an FAP that are not available for 
public disclosure are described in 21 CFR 571.1(h)(2).  Food additive regulations are not 
proprietary, which means that any manufacturer that can produce the additive can sell the 
additive for the intended use. 
 
Please note that, in addition  to FDAs requirements, you should contact other Federal 
Government agencies such as the Occupational Health and Safety Administration 
(OSHA) and the Environmental  Protection Agency (EPA) regarding actions that must be 
taken pursuant to other statutes prior to use of the food additive. 
 
The general format required for an FAP, as outlined  in 21 CFR 571.1(c), is as follows: 
 
  Name and all pertinent information  concerning the food additive,  including 
chemical identity and composition  of the additive or manufacturing  methods and 
controls if the chemical identity  and composition  are not known; 
  intended use, use level (amount of the food additive proposed for use), and 
labeling  (cautions, warnings, shelf life, directions for use);  
  data establishing  the intended effect (physical, nutritional,  or other technical 
effect); 
  a description  of analytical methods to determine the amount of the food additive 
in the food; 
  safety evaluation; 
  proposed tolerances for the food additive; 
  proposed regulation; and 
  environmental  assessment. 
 
C. Identity and Composition 
 
  1. Identity 
 
This section of the FAP must include information  which identifies  and characterizes the 
food additive,  including  the composition  of the food additive.  For chemicals intended for 
use as food additives this information  should include:  (1) formal chemical name; (2) 
common names, synonyms, or trade names; (3) Chemical Abstracts Service (CAS) 
registry number; (4) empirical and structural formulae, and molecular or formula weights.  
Information characterizing the chemical, biological,  and physical properties that can be 
used to identify the food additive must be submitted as well. 
 
For a food additive that is a mixture of substances (mixture),  the FAP should identify 
as many of the component substances (components) in the mixture as needed to 
adequately define the composition  of that food additive.  The minimum  amount of a 
component in a food additive  that is a mixture, that may need to be identified  as a 
component in the food additive is determined on a case-by-case basis. In addition,  the 
FAP should provide the composition  of each component in the mixture. 
6  Contains Nonbinding Recommendations 
Material balance calculations4 for the manufacturing  process of the food additive should 
be provided.  
 
For food additives of natural origin,  additional  information  describing the source of the 
additive should be included in order to identify  the additive.  Additional  information 
describing the source of food additives of natural origin can include the systematic name, 
genus and species, variability  based on climate, or other geographical  factors. 
 
Because of the potential variability  in the strength, purity, and quality of food additives of 
natural origin,  more information  may be needed to identify these food additives than may 
be needed for other food additives. In an FAP, the information  used to identify  a food 
additive of natural origin must be sufficient to ensure clarity about the identity  of the food 
additive in the food additive regulation. 
 
Food additives of natural origin  can be similar  to other food additives that are mixtures in 
that they may contain more than one component.  For a food additive that is a mixture of 
components, information  about the identities of the components and their presence and 
amounts in the mixture ensures consistency in the identity  and composition  of the food 
additive.  Similarly,  for a food additive of natural origin, the identity and composition  of 
the food additive should be consistent. 
 
Users of food additives expect food additives to be exactly what they are claimed to be in 
terms of identity.   The level of detail needed to identify  a food additive of natural origin 
depends on the nature of the food additive and is determined on a case-by-case basis. 
 
In accordance with 21 CFR 571.1(c) A, where such information  is not available,  a 
statement as to the reasons why it is not available should be submitted. 
 
FDA recommends that you include additional  relevant information  depending on the 
specific food additive.  For example, the characterization for an enzyme should include 
the systematic name, enzyme source, enzymatic activity, substrate specificity,  amino acid 
sequence, enzyme kinetics (temperature and pH), and structural modifications  introduced 
chemically  or genetically.  Appendix 1 of this document provides additional  guidance for 
additives that are produced by fermentation and/or biotechnological  processes using cell 
culture, bioengineered  microorganisms,  or bioengineered  plants.  This guidance does not 
cover genetically  engineered animals. For information  about the regulation of genetically 
engineered animals, see FDAs GFI #187 entitled Regulation  of Genetically  Engineered 
Animals Containing  Heritable Recombinant DNA Constructs.i 
   
                                              
4 For the purposes of this guidance, "material balance calculation" means a mathematical 
comparison between the amounts of material inputs and the amounts of material outputs of a 
process system. 
7  Contains Nonbinding Recommendations 
  2. Manufacturing methods and controls 
 
  a. Manufacturing process 
 
When the chemical identity and composition  of the food additive is not known, you must 
provide information  in sufficient detail to permit the evaluation  of the method of 
manufacture and the analytical controls used during the various stages of manufacturing, 
processing or packaging of the food additive (21 CFR 571.1(c)). In practice, an FAP will 
most likely  be deficient without a full and complete description of the methods used in 
the manufacturing process of the food additive and FDA will request this information 
from any sponsor who does not supply it with the FAP. Accordingly,  FDA recommends 
that you include this information  in your FAP under all circumstances. 
 
This section should contain a full description  of the methods used in the manufacture and 
processing of the food additive.  This description  should provide sufficient detail to allow 
CVM to evaluate the ability  of the process(es) to produce a consistent food additive with 
specified strength, purity, and quality.   For example, a complete description  should 
include:  (1) a list of all solvents, catalysts, and reactive ingredients  used; the quantities of 
these used; and the order of addition  of these chemicals during production;  (2) times, 
temperatures, and pH conditions  for all steps in the manufacturing process; (3) 
identification  of all manufacturing control points; and (4) identification  of all analytical 
methods used to monitor chemical reactions during manufacture, including  information 
that demonstrates that the methods perform within appropriate limits.   If manufacture of 
the food additive calls for extraction, separation, and/or purification  steps, the starting 
material should be adequately identified  and all methods and materials used to extract, 
separate, and/or purify the food additive should be provided.  You should also identify 
any alternative methods you may use in the manufacture of the food additive and provide 
as full a description  as possible for the alternative methods. 
 
A complete description  of the packaging process of the food additive,  including  the 
containers used and the final products labeling  (see section D below) should be 
provided. Dependent on the identity  of the food additive, the packaging operation should 
be validated, and both theoretical and actual packaging yields should be provided.  If the 
food additive is to be further processed with other ingredients or carriers into an animal 
food premix or supplement, a complete description of the manufacturing process to 
accomplish this should be provided.  Each ingredient  and carrier should be described by 
its chemical name, appropriate analytical tests, and specifications.  The final packaged 
additive should also be described by a set of appropriate tests and specifications.  These 
data and information  should be consistent with the information  present in the additives 
labeling. 
   
8  Contains Nonbinding Recommendations 
  b. Batch preparation 
 
When fermentation processes are used, all instructions  for manufacturing batch 
preparations should be given.  Appendix 1 provides additional  guidance for additives 
produced by fermentation processes. This should include procedures for preparing and 
maintaining  the working cell banks. 
 
  c. Specifications  for chemical identity  and purity 
 
To ensure that a food additive is of consistent chemical identity and purity, the FAP 
should describe the final food additive by a set of chemical, physical, and/or biological 
properties and specifications prescribing  the minimum  content of the desired 
component(s) and identifying  and limiting  any reaction byproducts and other impurities.   
The food additives specifications and their ranges should be based on analyses of three 
to five production  batches.  If it is not possible to base specifications and their ranges on 
analyses of production  batches, then the food additive specifications can be based on 
analyses of a significant  number of pilot production  batches.  The number of pilot 
production  batches needed to determine the food additive specifications is based on the 
variability  observed between the pilot production batches during manufacturing. Raw 
data and statistical analyses of the data should be submitted. 
 
The specifications can include,  for example, physical form, color, minimum  content of 
the desired component, ash content, moisture content, maximum content of reaction 
byproducts, and limits for impurities  and contaminants. 
 
  d. Food additive stability  and mixability 
 
The FAP must include stability  data (21 CFR 571.1(c) A).  Stability  testing should be 
conducted under the intended conditions  of use of the food additive over its anticipated 
lifetime.  Data from stability  studies should be submitted for three production  batches.  If 
using pilot  batches in place of production  batches, you should consult the Division  of 
Animal Feeds regarding stability  testing. The data should be fitted to a least squares 
regression, with the upper and lower 95% confidence limits  shown.  Where regression 
analysis is not possible, analysis of variance is an appropriate statistical analysis method.  
The analytical method used to determine stability  should be validated in the different 
animal food types that the food additive is intended to be used.  Stability  should be 
demonstrated for the food additive as packaged alone (e.g., as a final product with 
diluents or carriers) and for the food additive mixed in a representative animal food 
matrix (e.g., complete food or vitamin  or mineral premix) for the projected time the 
additive and the animal food containing  the additive will be available for sale.  If stability 
testing is to be performed on pelleted, canned, or liquid  animal food, then the time and 
temperature for pelleting,  canning, and liquid  food conditions,  respectively, should be 
provided in the FAP.  If needed to ensure the identity,  strength, quality,  or purity,  an 
expiration  date must be proposed (21 CFR 571.1(c) A). 
 
9  Contains Nonbinding Recommendations 
Many food additives are added to animal food at concentrations in the parts per million  or 
even parts per billion  levels, and it is essential that these additives can be homogeneously 
mixed in the animal food.  When necessary, the FAP should provide sufficient data and 
information  to demonstrate that the additive can be homogenously  mixed in a 
representative food matrix, using mixing  equipment that is readily available to a finished 
feed manufacturer.  That information  should include the coefficient of variation.  The 
final concentration of the additive in the intended food matrix will determine the 
acceptable amount of variability. 
 
D. Intended Use and Use Level, and Labeling 
 
  1. Intended use and use level 
 
In accordance with 21 CFR 571.1(c) C, a petition must include data establishing  that the 
food additive will have its intended physical or other technical effect or that it may 
reasonably be expected to become a component, or to affect the characteristics, directly 
or indirectly,  of food and the amount necessary to accomplish this. This section of the 
FAP should include a discussion  of the intended use and use level of the food additive. 
Such a discussion should include:  (1) the type of animal food in which the additive will 
be used; (2) use level of the additive to be used in each type of animal food; (3) proposed 
maximum  and typical use levels expressed as a concentration by weight; (4) a clear 
statement of the intended effect(s) of the additive; and (5) the species, age, and life stage 
of animals that will consume the food additive. 
 
A food additive may have more than one intended effect.  Some examples of intended 
effects of food additives include the following  attributes: nutrient, aroma/flavor, taste, 
soluble or insoluble  fiber, stabilization,  emulsification,  sequestration, preservation, anti-
oxidant activity, anti-caking,  or another technical effect.  A list of terms that describe 
some physical or technical functional  effects of types of food additives is provided in 21 
CFR 170.3(o). 
 
If the food additive is one for which a tolerance, of any kind,  is required in order to 
ensure its safety, the proposed use level should be no higher than the minimum  level 
required to accomplish the intended effect, even though safety data may a support higher 
tolerance (21 CFR 571.1(c) B). In other words, for these food additives,  the proposed use 
level is determined based on the minimum  level required to accomplish the intended 
effect and not based on the maximum  tolerance level. 
 
We advise you to discuss the intended use of the food additive with CVM early in the 
development process.  Contact information  for the Division  of Animal Feeds is provided 
in section III.A of this document. 
   
10  Contains Nonbinding Recommendations 
  2. Labeling 
 
As specified in 21 CFR 571.1(c) B, you are required to submit copies of the labeling 
proposed for the food additive.  The labeling  is required to be in compliance with 21 CFR 
part 501.  You should consult with CVM regarding the appropriate content and format of 
the labeling.   In general, a food additive label should include the following:  1) the 
product name or brand name; 2) description of the purpose including  the types of animal 
food to which it can be added; 3) list of all ingredients,  including  inert compounds,  listed 
by their common or usual name in descending order of predominance according to 
weight in the marketed product; 4) directions for use, including  caution or warning 
statements; 5) the name and principal  mailing  address of the manufacturer or distributor; 
6) product expiration statement (if needed based on the stability  testing discussed 
previously);  and 7) a declaration of net contents, which should appear in the bottom 30% 
of the label.  The labeling  is also required to include any other information  deemed 
necessary by FDA to ensure that the food additive  can be safely incorporated and used in 
animal food. 
 
E. Data Establishing the Intended Effect 
 
Data must be submitted to demonstrate that the food additive will have its intended effect 
and the amount of the food additive required to accomplish the intended effect (21 CFR 
571.1(c) C).  The number of studies and the detail and type of data required to 
demonstrate the intended effect will vary, depending upon the food additive and its 
intended use.  To demonstrate the minimum  level required to accomplish the intended 
effect, the intended use of the food additive should be evaluated at several levels above 
and below the proposed use level (for example, at half (12x) the intended use level, at the 
intended use level (1x), and at two times (2x) the intended use level).  The functionality 
experiments should be carefully designed, taking into account the need for controls.  
Additionally,  some food additives may have a technologically  self-limiting  use level; for 
example, where the food additive has a maximum  concentration in animal food above 
which the food becomes unpalatable to the animal.  In such a case, data are still  needed to 
demonstrate the intended effect of the additive and documentation  should be provided 
about the self-limiting  use level.  As the self-limiting  use level depends on characteristics 
of the food additive, the documentation  needed to describe the self-limiting  use level will 
depend on those characteristics. 
 
For general guidance on writing a study protocol to evaluate the intended use of the food 
additive,  you are advised to consult FDAs GFI #215 entitled Target Animal Safety and 
Effectiveness Protocol Development and Submission.ii   Analytical methods used to 
determine the intended effect of a food additive should be validated to ensure that they 
measure the appropriate parameter.  Validation  parameters should include variables, such 
as matrix effects. 
   
11  Contains Nonbinding Recommendations 
For additional  guidance on the types of information  that you can use to demonstrate the 
intended effect of a food additive,  we recommend that you consult FDAs GFI #53 
entitled Evaluation  of the Utility  of Food Additives in Diet Fed to Aquatic Animals.iii  
Although some of the recommendations  in GFI #53 pertain specifically to food additives 
for aquatic animals that are used at extremely low use levels, such as vitamins,  many of 
the recommendations are generally applicable to other types of food additives. 
 
F. Analytical Methods 
 
The petition  must include stability  data, and, if the data indicate that it is needed to ensure 
the identity,  strength, quality,  or purity of the additive, the expiration  date that will be 
employed (21 CFR 571.1(c) A). The stability  data should include a detailed description 
of the method(s) used to determine the strength, purity,  and quality of the food additive. 
All tests used to establish final food additive specifications should be validated with 
respect to correctness or be commonly  accepted methods, i.e., methods published  in the 
Official Methods of Analysis of the Association of Official Analytical Chemists, the 
Food Chemicals Codex, or the United States Pharmacopeia.  Such analytical methods 
should be described in sufficient detail and specificity to permit CVM to assess the 
capabilities  of the methods to determine the food additives strength, purity, quantity,  and 
quality as it is produced, after it is purified,  in its marketed and packaged matrix, and in 
representative animal food. The methods proposed must be practical methods that can be 
performed routinely  by trained laboratory personnel.  For general guidance on validating 
an analytical method, you are advised to consult FDAs GFI #63 entitled Validation  of 
Analytical Procedures: Definition  and Terminology  VICH GL1iv and GFI #64 entitled 
Validation  of Analytical Procedures: Methodology:  Final Guidance VICH GL2.v 
 
G. Safety Evaluation 
 
As stated in 21 CFR 571.1(c) E, the safety evaluation section of the petition must include 
full reports of investigations  with respect to the safety of the food additive.  Typically, 
safety studies (non-clinical  laboratory studies) are conducted in accordance with the 
Good Laboratory Practice (GLP) regulations (21 CFR part 58).  The FAP must include a 
statement that a safety study was conducted in compliance with GLP regulations or, if the 
study was not conducted in compliance with the GLP regulations,  a brief statement of the 
reason for the noncompliance  (21 CFR 571.1(k)).  You should explain what impact the 
lack of compliance with GLP regulations  has on any conclusions in such reports 
regarding the food additives safety. 
 
  1. Human food safety 
 
The FAP is required to have scientific data necessary for demonstrating  that the residues 
of the food additive are safe to humans consuming  edible products of animals fed the 
food additive.  See Section 409(c)(3)(A) of the FD&C Act.  The three areas below should 
be addressed in the FAP when the additive is intended for use in a food-producing  animal 
species.  In certain cases, the types of studies described in this and the other referenced 
guidance documents will not be appropriate, such as when a food additive will be used 
12  Contains Nonbinding Recommendations 
only in food products for companion animals.  We encourage you to contact CVM to 
discuss what types of safety information  should be included  for a particular food additive 
under the conditions  of its intended use. 
 
  a. Toxicology 
 
The toxicity  to human consumers potentially  associated with the use of a food additive 
can be assessed using a standard battery of toxicology  tests.  Each test is designed to 
examine a different toxicological  endpoint.  See FDAs GFI #3 entitled General 
Principles for Evaluating  the Safety of Compounds Used in Food-producing  Animals.vi 
 
We also recommend that you consult FDAs GFI #149 entitled Studies to Evaluate the 
Safety of Residues of Veterinary Drugs in Human Food: General Approach to Testing 
VICH GL33 for general toxicology  study recommendations.vii  You should consult the 
referenced guidance documents if you plan to design and conduct genotoxicity  studies,viii 
carcinogenicity  studies,ix reproduction studies,x repeat dose (90-day) toxicity  studies,xi 
developmental  toxicity  studies,xii or repeat dose (chronic) toxicity  studies.xiii  We may ask 
for additional  toxicology  testing as necessary to demonstrate that there is reasonable 
certainty of no harm with respect to potential residues in the edible tissues of animals that 
humans may consume. 
 
  b. Residue chemistry 
 
We recommend that you consult FDAs GFI #3 entitled General Principles for 
Evaluating  the Safety of Compounds Used in Food-producing  Animals.xiv  GFI #3 
provides a description of suggested residue chemistry studies to determine the amount of 
the food additive or substances/components that could be produced in or added to human 
food products derived from animals consuming the food additive.  GFI #3 also describes 
studies to characterize those residues.  These studies could include specially designed 
assays to characterize the distribution  and potential accumulation  of food additive 
residues, including  the parent molecule and its metabolites,  in the different tissues of 
food-producing  animals and in milk  or eggs.  You should consult the referenced guidance 
documents if you plan to design and conduct metabolism  studies to determine the 
quantity and identify  the nature of residues,xv laboratory animal comparative metabolism 
studies,xvi marker residue depletion  studies to establish additive withdrawal periods,xvii or 
validation  of analytical methods used in residue depletion  studies.xviii 
 
  c. Microbial  food safety 
 
If the food additive possesses measurable antimicrobial  activity,  such as antibiotics  used 
in distillers  grains production,  sufficient data should be submitted to determine whether 
the additive has any adverse impact on the emergence and development  of antimicrobial 
resistance among food-borne pathogens and related commensal bacteria in the intestinal 
tract of animals consuming  the food additive. In addition,  the impact on human intestinal 
flora should be assessed for such food additives to determine whether a microbiological 
acceptable daily intake (ADI) should be established for the food additive.  We 
13  Contains Nonbinding Recommendations 
recommend that you consult FDAs GFI #152 entitled Evaluating  the Safety of 
Antimicrobial  New Animal Drugs with Regard to Their Microbiological  Effects on 
Bacteria of Human Health Concernxix and FDAs GFI #159 entitled Studies to Evaluate 
The Safety of Residues of Veterinary Drugs in Human Food: General Approach to 
Establish a Microbiological  ADI.xx 
 
  2. Target animal safety 
 
The FAP will need to address the safety of the food additive under its intended conditions 
of use (21 CFR 571.1(c) E).  In most cases, a study should be conducted in the proposed 
target animal species to demonstrate safety.  Target animal safety studies should be 
conducted using the life stage and animal species for which the food additive will be 
marketed.  In cases where the food additive is intended for multiple  animal species or life 
stages, the food additive  should be tested in the most sensitive life stage and/or species.  
You should use current scientific literature to identify  the most sensitive life stage and/or 
species.  Prior to initiating  studies, you should consult with us to determine if your 
identification  of the most sensitive life stage and/or species is acceptable. 
 
Target animal safety studies should demonstrate the margin of safety for the intended use 
of the food additive.  When designing  safety studies in target animals, at least three 
inclusion  levels (treatments) of the food additive should be used.  In addition,  a 
concurrent control group of test animals should be included in the study.  Thus, the study 
design should include at least four experimental groups: a negative control, the maximum 
proposed use level (1X), and two multiples  of this level [in most cases three times (3X) 
and five times (5X)] for a period of time in excess of the recommended maximum 
duration of use.  However, alternative study designs can be considered where appropriate.  
Parameters for evaluation of safety in each group can include:  clinical  observations (e.g., 
of behavior, appearance, and eating patterns), blood analyses, mortality,  weight gain, feed 
intake, and necropsy findings  (gross and histopathologic  abnormalities).   We also 
recommend that you consult FDAs GFI #185 entitled Target Animal Safety for 
Veterinary Pharmaceutical Products VICH GL43xxi for general target animal safety 
study recommendations. The number of animals used in a study to evaluate the safety of 
a food additive should be based on the expected variation in the parameters that will be 
measured, i.e., the study should have sufficient animal numbers such that the power of 
the study is adequate to ensure that any treatment-related effects can be detected. 
 
CVM requests that you contact us early in protocol development  in order to determine 
how target animal safety studies can be completed without terminal end points. 
 
We recommend you discuss target animal safety study designs with CVM early in the 
development process and then submit study protocols for CVM review and concurrence 
prior to initiating  any target animal safety studies.  For general guidance for writing 
protocols, consult FDAs GFI #215 entitled Target Animal Safety and Effectiveness 
Protocol Development and Submission.xxii 
   
14  Contains Nonbinding Recommendations 
H. Proposed Tolerances for the Food Additive 
 
Tolerances for food additives fed to animal species used for human food represent the 
maximum  concentration of the additive permitted in edible tissues (for example, meat, 
milk,  and eggs).  Tolerances are established based upon residues of the food additive in 
edible tissues of food-producing  animals that consumed the additive.  Tolerances are 
intended to ensure that residues of food additives in edible tissues will have no harmful 
effects when consumed by people.  The potential for establishment of a tolerance is a 
facet of the human food safety evaluation discussed earlier. 
 
I. Proposed Regulation 
 
We recommend that you provide a proposed regulation  as part of your FAP.  This section 
of the FAP should contain a draft food additive regulation  in which you describe the 
conditions  under which the additive can be safely used in animal food.  The proposed 
regulation  should contain information  adequate to ensure safety of the additive under 
each and every condition  of use to be permitted and should include any restrictions, such 
as animal species limitations,  that are important to ensure safe use of the food additive.  
Special attention should be given to food additive specifications,  and/or any other 
additional  information  that is needed for the identification  and safe use of the food 
additive under the conditions  of its intended use.  Current approved animal food additives 
are found in 21 CFR part 573. 
 
J. Environmental Assessment 
 
FDA is required under the National Environmental  Policy Act (NEPA) to consider the 
environmental  impact of its regulatory actions, including  those relating to food additives.  
FDAs regulations in part 25 (21 CFR part 25) set forth procedures to supplement the 
NEPA regulations of the Council on Environmental  Quality (CEQ) under 40 CFR parts 
1500-1508.  In accordance with 21 CFR 25.20(i), approval of an FAP and approval of a 
request for an exemption for investigational  use of a food additive are both actions that 
ordinarily  require preparation of at least an Environmental  Assessment (EA), unless the 
action is of a class that qualifies for categorical exclusion.  An EA is a concise public 
document that provides sufficient evidence and analysis for CVM to determine whether a 
Finding  Of No Significant  Impact (FONSI) can be prepared or an Environmental  Impact 
Statement (EIS) is necessary (21 CFR 25.40).  In accordance with 21 CFR 571.1(c) H 
and 21 CFR 25.15(a), you are required to submit either a claim for categorical exclusion 
from the requirement to prepare an EA, or an EA under 21 CFR 25.40.  In addition,  21 
CFR part 25 requires petitions for food additives to include sufficient information  to 
allow the FDA to assess whether environmental  impacts may occur from the use and 
disposal of the food additive. 
   
15  Contains Nonbinding Recommendations 
A categorical exclusion from the requirement to prepare an EA is defined as a category of 
actions which do not individually  or cumulatively  have a significant  effect on the human 
environment  (40 CFR 1508.4).  The classes of actions for food additives that typically 
qualify for a categorical exclusion  are listed under 21 CFR 25.32.  To claim a categorical 
exclusion from the requirement to prepare an EA you must state in the petition  that the 
intended use qualifies  for a categorical exclusion,  cite the particular categorical exclusion 
that is claimed, and state that to your knowledge, no extraordinary circumstances exist 
(21 CFR 25.15(a) and (d)).  FDA will review the petition and determine whether the 
categorical exclusion is applicable and whether any extraordinary circumstances exist 
that indicate the intended use of the additive may significantly  affect the quality  of the 
human environment. 
 
Some examples of extraordinary circumstances are described in 21 CFR 25.21.  These 
extraordinary circumstances include any actions for which the available data establish 
that there is potential for serious harm to the environment at the expected exposure level 
(21 CFR 25.21(a)).  Other extraordinary circumstances include actions that may 
adversely affect a species (flora or fauna) or the critical habitat of a species that is entitled 
to special protection under a Federal law such as the Endangered Species Act or under 
the Convention  on International Trade in Endangered Species of Wild Flora and Fauna 
(21 CFR 25.21(b)).  Additional  extraordinary circumstances are described in the CEQ 
regulations implementing  NEPA (40 CFR 1508.27). 
 
When an EA is necessary, information  concerning the purpose and scope of information 
that you must submit for an EA is contained in 21 CFR 25.40.  Data included in the EA 
can be obtained from the scientific literature and from adequate and controlled  laboratory 
studies. 
 
FDA will evaluate the information  contained in the EA to determine whether it is 
accurate and objective and whether the proposed action may significantly  affect the 
quality of the human environment  (21 CFR 25.15(b)).  If significant  effects requiring the 
preparation of an EIS are identified,  then FDA will prepare an EIS.  If such effects are 
not identified,  then FDA will prepare a FONSI. 
 
We recommend that you contact us early in the development of the FAP so that FDA can 
determine whether the food additive qualifies  for a categorical exclusion or to determine 
the scope of the EA if one is needed.  Contact information  for the Division  of Animal 
Feeds is provided in section III.A of this document. 
   
16  Contains Nonbinding Recommendations 
IV. OTHER INFORMATION 
 
In accordance with 21 CFR 571.1(a), the FAP must be submitted in triplicate.  However, 
when submitting  large data files and statistical analyses, a computer compact disk (CD) 
can be included  instead of sending two of the three paper copies.  Furthermore, FDA 
regulations at 21 CFR 571.1(c) require the information  in an FAP be submitted by 
organizing  it according to the lettered headings noted in 21 CFR 571.1(c).  If any of the 
data or information  is in a foreign language, accurate and complete English translations 
must be provided (21 CFR 571.1(a)). 
 
17  Contains Nonbinding Recommendations 
APPENDIX 1 
 
Food additives used in animal food can be obtained from fermentation or 
biotechnological  processes, including  bioengineered  microorganisms  and plants.  For 
these food additives, the FAP should include a complete description of the manufacturing 
process including  complete descriptions of the following: 
 
1)  Systematic name (Genus species) and source of each organism or cell line used to 
produce the food additive. 
2)  Any changes made to the organism and how those changes are accomplished. 
3)  The process used to select or induce cell lines as producers of the food additive. 
4)  Any recombinant techniques used on the organism or cell line, including  a complete 
description of the bioengineering  process (e.g., construction of the expression 
cassettes and the vectors, insertion technique, etc.). 
5)  Any traditional  mutagenic techniques used on the organism or cell line. 
6)  The processes and procedures used to maintain organism integrity in response to 
genetic drift and selection pressure. 
7)  How the master cell and working cell banks are stored, maintained,  and inoculums 
are transferred. 
8)  The media (e.g., fermentation broth) used to cultivate or grow the organism or cell 
culture.  Each ingredient in the media should be identified  by appropriate analytical 
tests and specifications.  For other organisms, include a complete description of how 
the organism is produced and maintained. 
9)  The production  or fermentation process, including  temperature, time, and pH of the 
process and all controls that are used to maintain the integrity  of the reactor and the 
media, pre-, post- and during fermentation or production. All analytical control points 
and methods or process analytical controls used to monitor the manufacturing process 
should be identified. 
10)  The extraction and purification  processes, including  the identity  and composition  of 
all solvents, precipitants,  emulsifiers,  and other agents added during these processes. 
11)  The processes and products used to sterilize the reactors. 
12)  The steps taken upon discovery of contamination. 
 
   
18  Contains Nonbinding Recommendations 
ENDNOTES 
 
                                              
i GFI #187, Regulation of Genetically Engineered Animals Containing Heritable Recombinant 
DNA Constructs 
http://www.fda.gov/downloads/AnimalVeterinary/GuidanceComplianceEnforcement/Guidancefo
rIndustry/UCM113903.pdf 
ii GFI #215, Target Animal Safety and Effectiveness Protocol Development and Submission 
http://www.fda.gov/downloads/AnimalVeterinary/GuidanceComplianceEnforcement/Guidancefo
rIndustry/UCM241787.pdf 
iii GFI #53, Evaluation of the Utility of Food Additives in Diet Fed to Aquatic Animals 
http://www.fda.gov/AnimalVeterinary/GuidanceComplianceEnforcement/GuidanceforIndustry/u
cm053413.htm 
iv GFI #63, Validation of Analytical Procedures: Definition and Terminology 
http://www.fda.gov/downloads/AnimalVeterinary/GuidanceComplianceEnforcement/Guidancefo
rIndustry/UCM052377.pdf 
 
v GFI #64, Validation of Analytical Procedures: Methodology 
http://www.fda.gov/downloads/AnimalVeterinary/GuidanceComplianceEnforcement/Guidancefo
rIndustry/UCM052379.pdf 
 
vi GFI #3, General Principles for Evaluating the Safety of Compounds Used in Food-Producing 
Animals  
http://www.fda.gov/downloads/AnimalVeterinary/GuidanceComplianceEnforcement/Guidancefo
rIndustry/UCM052180.pdf 
vii GFI #149, Studies to Evaluate the Safety of Residues of Veterinary Drugs in Human Food: 
General Approach to Testing VICH GL33 
http://www.fda.gov/downloads/AnimalVeterinary/GuidanceComplianceEnforcement/Guidancefo
rIndustry/UCM052521.pdf 
viii GFI #116, Studies to Evaluate the Safety of Residues of Veterinary Drugs in Human Food: 
Genotoxicity Testing VICH GL23 
http://www.fda.gov/downloads/AnimalVeterinary/GuidanceComplianceEnforcement/Guidancefo
rIndustry/UCM052656.pdf 
ix GFI #141, Studies to Evaluate the Safety of Residues of Veterinary Drugs in Human Food: 
Carcinogenicity Testing VICH GL28 
http://www.fda.gov/downloads/AnimalVeterinary/GuidanceComplianceEnforcement/Guidancefo
rIndustry/UCM052527.pdf 
x GFI #115, Safety Studies for Veterinary Drug Residues in Human Food: Reproduction 
Toxicity Testing VICH GL22 
http://www.fda.gov/downloads/AnimalVeterinary/GuidanceComplianceEnforcement/Guidancefo
rIndustry/UCM052655.pdf 
 
19  Contains Nonbinding Recommendations 
                                                                                                                                        
xi GFI #147, Studies to Evaluate the Safety of Residues of Veterinary Drugs in Human Food: 
Repeat-Dose (90-Day) Toxicity Testing VICH GL31 
http://www.fda.gov/downloads/AnimalVeterinary/GuidanceComplianceEnforcement/Guidancefo
rIndustry/UCM052523.pdf 
xii GFI #148, Studies to Evaluate the Safety of Residues of Veterinary Drugs in Human Food: 
Developmental Toxicity Testing VICH GL32 
http://www.fda.gov/downloads/AnimalVeterinary/GuidanceComplianceEnforcement/Guidancefo
rIndustry/UCM052522.pdf 
xiii GFI #160, Studies to Evaluate the Safety of Residues of Veterinary Drugs in Human Food: 
Repeat-Dose (Chronic) Toxicity Testing VICH GL37 
http://www.fda.gov/downloads/AnimalVeterinary/GuidanceComplianceEnforcement/Guidancefo
rIndustry/UCM052505.pdf  
xiv GFI #3, General Principles for Evaluating the Safety of Compounds Used in Food-Producing 
Animals  
http://www.fda.gov/downloads/AnimalVeterinary/GuidanceComplianceEnforcement/Guidancefo
rIndustry/UCM052180.pdf 
xv GFI #205, Studies to Evaluate the Metabolism and Residue Kinetics of Veterinary Drugs in 
Food-producing Animals: Metabolism Study to Determine the Quantity and Identify the Nature 
of Residues (MRK) VICH GL46 
http://www.fda.gov/downloads/AnimalVeterinary/GuidanceComplianceEnforcement/Guidancefo
rIndustry/UCM207939.pdf 
xviGFI #206, Studies to Evaluate the Metabolism and Residue Kinetics of Veterinary Drugs in 
Food-Producing Animals: Comparative Metabolism Studies in Laboratory Animals VICH GL47 
http://www.fda.gov/downloads/AnimalVeterinary/GuidanceComplianceEnforcement/Guidancefo
rIndustry/UCM207940.pdf  
xvii GFI #207, Studies to Evaluate the Metabolism and Residue Kinetics of Veterinary Drugs in 
Food-Producing Animals: Marker Residue Depletion Studies to Establish Product Withdrawal 
Periods VICH GL48 
http://www.fda.gov/downloads/AnimalVeterinary/GuidanceComplianceEnforcement/Guidancefo
rIndustry/UCM207941.pdf 
xviii GFI #208, Studies to Evaluate the Metabolism and Residue Kinetics of Veterinary Drugs in 
Food-Producing Animals: Validation of Analytical Methods used in Residue Depletion Studies 
VICH GL49 
http://www.fda.gov/downloads/AnimalVeterinary/GuidanceComplianceEnforcement/Guidancefo
rIndustry/UCM207942.pdf 
xixGFI #152, Evaluating the Safety of Antimicrobial New Animal Drugs with Regard to Their 
Microbiological Effects on Bacteria of Human Health Concern 
http://www.fda.gov/downloads/AnimalVeterinary/GuidanceComplianceEnforcement/Guidancefo
rIndustry/UCM052519.pdf 
 
20  Contains Nonbinding Recommendations 
                                                                                                                                        
xx GFI #159, Studies to Evaluate the Safety of Residues of Veterinary Drugs in Human Food: 
General Approach to Establish a Microbiological ADI VICH GL36(R)   
http://www.fda.gov/downloads/AnimalVeterinary/GuidanceComplianceEnforcement/Guidancefo
rIndustry/UCM124674.pdf 
xxi GFI #185, Target Animal Safety for Veterinary Pharmaceutical Products VICH GL43 
http://www.fda.gov/downloads/AnimalVeterinary/GuidanceComplianceEnforcement/Guidancefo
rIndustry/UCM052464.pdf 
xxiiGFI #215, Target Animal Safety and Effectiveness Protocol Development and Submission 
http://www.fda.gov/downloads/AnimalVeterinary/GuidanceComplianceEnforcement/Guidancefo
rIndustry/UCM241787.pdf 
21