#262 
 
Pre-Submission Consultation Process for 
Animal Food Additive Petitions or 
Generally Recognized as Safe (GRAS) 
Notices 
 
Guidance for Industry 
 
Submit comments on this guidance at any time.  Submit electronic comments to 
https://www.regulations.gov.  Submit written comments to the Dockets Management Staff 
(HFA-305), Food and Drug Administration, 5630 Fishers Lane, Rm. 1061, Rockville, MD 
20852.  All comments should be identified with docket number FDA-2020-D-0064. 
 
For further information regarding this document, contact AskCVM@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, 7500 Standish 
Place, Rockville MD 20855, and may be viewed on the Internet at either 
https://www.fda.gov/animal-veterinary, https://www.fda.gov/regulatory-information/search-fda-
guidance-documents, or https://www.regulations.gov. 
 
 
 
U.S. Department of Health and Human Services 
Food and Drug Administration 
Center for Veterinary Medicine  
December 2020   
  Contains Nonbinding Recommendations 
 
Table of Contents 
 
I.  Introduction .......................................................................................................................... 3 
II.  Background .......................................................................................................................... 4 
III.  Pre-Submission Consultation Offered by FDA ................................................................. 6 
A.  Pre-Submission Consultation Meetings and General Correspondence Requests .... 7 
B.  Investigational Food Additive (IFA) File for Animal Food ....................................... 7 
1.  Request for Review of a Study Protocol ..................................................................... 8 
2.  Request for Authorization for Use of Animal Products in Human or Animal Foods 
(FUA Request) ........................................................................................................... 9 
3.  Request for Review of the Adequacy of Existing Data from a Completed Study ....... 9 
C.  What to Include in a Request to Establish an IFA File ............................................ 10 
D.  What to Include in Each Submission Type to an Established IFA File .................. 11 
1.  Request for Review of a Study Protocol ................................................................... 11 
2.  Request for Authorization for Use of Animal Products in Human or Animal Foods 
(FUA Request) ......................................................................................................... 12 
3.  Request for Review of the Adequacy of Existing Data from a Completed Study ..... 13 
E.  How to Submit Information to an IFA File ............................................................... 14 
IV.  FDA Response to Requests Submitted to an IFA File .................................................... 14 
V.  Environmental Assessment ............................................................................................... 14 
VI.  Freedom of Information Act ............................................................................................. 16 
Appendix 1  Table of Abbreviations and Acronyms Used in this Guidance ....................... 17 
Appendix 2  Study Protocol Content for a Study of a Substance Intended for 
Investigational Use in Animal Food .................................................................. 18 
Appendix 3  Investigational Animal Food Substance Final Study Report Format ............ 25 
 
   
  2  Contains Nonbinding Recommendations 
 
Pre-Submission Consultation Process for Animal Food Additive 
Petitions or Generally Recognized as Safe (GRAS) Notices 
 
Guidance for Industry 
 
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 the Food and Drug Administrations (FDAs) 
Center for Veterinary Medicine (CVM) recommends be included in:  
(1) pre-petition consultations prior to submission of food additive petitions (FAP) for food 
additives intended for use in animal food; 
(2) pre-submission consultations regarding an animal food substance1 for which an entity 
plans to provide notice of its conclusion that the intended use of the substance is 
generally recognized as safe (GRAS) under FDAs animal food GRAS Notification 
program; or 
(3) a Food Use Authorization (FUA) request to permit the use, in human or animal foods, of 
animal products derived from animals that have been administered an investigational 
substance2 intended for use in animal food. 
 
This guidance is intended to help the petitioner, notifier, requestor, firm, stakeholder, or entity 
(collectively, you) submit information for effective and efficient pre-submission consultations 
for an FAP or GRAS substance and preparation of an FUA request.  This guidance also provides 
detailed information that will help you navigate FDAs current process for pre-submission 
consultations and FUA requests. 
 
This guidance also is intended to be used in conjunction with other FDA Guidance for Industry 
(GFI) documents that are cited in this document.  In general, FDAs guidance documents do not 
establish legally enforceable responsibilities.  Instead, guidances describe the Agencys current 
thinking on a topic and should be viewed only as recommendations, unless specific regulatory or 
statutory requirements are cited.  The use of the word should in Agency guidances means that 
something is suggested or recommended, but not required. 
 
                                                 
1 For this guidance, the term substance refers to a proposed use of a substance that may require the submission of 
an FAP or be the subject of a GRAS conclusion. 
2 For this guidance, the term investigational substance refers to a substance that is the subject of a scientific study 
to support a food additive petition or a GRAS conclusion. 
  3  Contains Nonbinding Recommendations 
 
II.  Background  
 
FDA regulates food under the Federal Food, Drug, and Cosmetic Act (FD&C Act).  Section 
201(s) of the FD&C Act (21 U.S.C. 321(s)) defines a food additive, in part, as any substance 
the intended use of which results or may reasonably be expected to result, directly or indirectly, 
in its becoming a component or otherwise affecting the characteristics of any food.  Section 
201(s) excludes from this definition any substance that is generally 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 (for example, new animal drugs or color 
additives).3 
 
On August 14, 2018, the President signed the Animal Drug and Animal Generic Drug User Fee 
Amendments of 2018 into law (Pub. L. 115-234).  This law includes a requirement that FDA 
publish a guidance relating to the voluntary pre-petition consultation process for food additives 
intended for use in animal food, including best practices for pre-petition submissions of study 
protocols and existing data (see section 306(c) of Pub. L. 115-234).  Issuance of this guidance is 
intended to fulfill that requirement. 
 
Food additives require pre-market approval based on data demonstrating safety and addressing 
the effect the additive is intended to produce.  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 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 (see section 
301(a) of the FD&C Act (21 U.S.C. 331(a))).  Therefore, for a firm to lawfully use a food 
additive in animal food, FDA must have issued a food additive regulation prescribing the 
conditions under which the additive may be safely used, and the additive must be used in 
accordance with the regulation (see section 409(a)(2) of the FD&C Act).  Section 409(b) through 
(g) of the FD&C Act and FDAs regulations located at part 571 in Title 21 of the Code of 
Federal Regulations (21 CFR part 571) provide for a petition process to establish that a use of an 
animal food additive is safe.  You may find additional information on CVMs website, 
Ingredients and Additives.4 
  
FDA recommends that prior to submitting an FAP, you verify that the substance is not already 
approved as an animal food additive for your proposed intended use.  FDA publishes animal 
                                                 
3 Under section 201(s) of the FD&C Act, the following types of substances are excluded from being animal food 
additives: (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 (that is, approval granted 
with respect to use of a substance in food prior to the amendment of the FD&C Act by the Food Additives 
Amendment of 1958 on September 6, 1958, by the FDA or the United States Department of Agriculture (USDA) 
pursuant to the FD&C Act, the Poultry Products Inspection Act, or the Meat Inspection Act (21 CFR 570.3(l)); and 
(5) new animal drugs.  
4 https://www.fda.gov/animal-veterinary/animal-food-feeds/ingredients-additives. 
 
  4  Contains Nonbinding Recommendations 
 
food additive regulations at 21 CFR parts 573 and 579.5  Furthermore, a particular use of a 
substance may not require an FAP because that use is GRAS.  FDA has affirmed certain 
substances as GRAS for their intended use in animal food and these are listed in 21 CFR parts 
582 and 584.  
 
The Current Animal Food GRAS Notices Inventory6 (Inventory) on FDAs website lists filed 
notices in which a firm has notified FDA of its conclusion that the notified substance is generally 
recognized, among qualified experts, to be safe under the conditions of its intended use and also 
provides FDAs response to each notice.  The listings in the inventory apply to the specific firm 
that made the safety conclusion of the use of a substance.  However, there is no provision in the 
FD&C Act or FDA regulations providing exclusivity for a substance that is used in food based 
on a conclusion that the substance is GRAS under the conditions of its intended use.  If you rely 
on a GRAS notice submitted by another person, you should carefully consider whether its 
production process, and/or the intended conditions of use of the notified substance, fall within 
the parameters and specifications addressed by the submitted GRAS notice.7  For more 
information about general recognition of safety and the GRAS notification program for animal 
food, please see 21 CFR part 570, subpart E, GRAS guidance documents, and FDAs website, 
Generally Recognized as Safe (GRAS) Notification Program.8  
 
When comparing your proposed use of a substance to existing animal food additive regulations, 
GRAS affirmation regulations, or GRAS conclusions in the inventory, bear in mind that a food 
additive regulation, GRAS affirmation, or GRAS conclusion is not for the substance itself, but 
for the intended use of the substance.  Therefore, you may be required to establish a food 
additive regulation or make a GRAS conclusion if your proposed intended use for a substance is 
different. 
 
If you intend to submit an FAP or want to participate in the GRAS notification program, FDA 
encourages pre-petition or pre-notification consultations with FDA, which could include 
meetings in person, teleconference calls, or video conferences to facilitate the development of 
information needed for an FAP or a GRAS notice, particularly for those who do not have 
previous experience preparing an FAP or GRAS notice for animal food.  FDA also recommends 
that you contact FDA early in the development process.  A meeting in person, teleconference 
call, or video conference with FDA staff to discuss a research plan for the use of a substance is 
often helpful for you to prepare an FAP or a GRAS notice.  Once a research plan is developed, 
FDA strongly suggests that you submit study protocols for FDAs review and receive comments 
before initiating the studies.  
 
                                                 
5 Food intentionally subjected to radiation is adulterated, unless the use of the radiation was in conformity with a 
current food additive regulation or an exemption (see section 402(a)(7) of the FD&C Act). 
6 https://www.fda.gov/animal-veterinary/generally-recognized-safe-gras-notification-program/current-animal-food-
gras-notices-inventory. 
7 Substances Generally Recognized as Safe, 81 FR 54960 at 54984 and 55013 (Aug. 17, 2016).  Available at 
https://www.govinfo.gov/content/pkg/FR-2016-08-17/pdf/2016-19164.pdf. 
8 All available at https://www.fda.gov/animal-veterinary/animal-food-feeds/generally-recognized-safe-gras-
notification-program. 
  5  Contains Nonbinding Recommendations 
 
III.  Pre-Submission Consultation Offered by FDA  
 
FDA offers pre-submission consultation to help facilitate the development of FAPs or GRAS 
notices for substances intended for use in animal food.  Pre-submission consultations are 
encouraged, but not required.  A pre-submission consultation may be used to: 
  discuss your proposed intended use of a substance and your development plan to 
support the use of the substance through an FAP or a GRAS conclusion based on the 
nature of the substance, its intended use, and the type of data available; 
  verify whether the quality and quantity of existing information is likely to meet the 
requirements for an FAP or a GRAS notice;   
  address questions about how to establish safe conditions of use of a substance;  
  consult with FDA in situations where there are uncertainties about how data may be 
interpreted and when those uncertainties are of such magnitude that they may affect 
the outcome of the overall safety determination regarding the use of a substance in 
animal food (for example, if different but reasonable interpretations of data in a 
bioassay could change the conclusion regarding the likely toxicity or carcinogenicity 
of the substance); 
  seek FDA input and recommendations on proposed studies and research 
methodologies intended to address the safe use of a substance in animal food;  
  determine the types of additional data and information that may be needed to meet the 
requirements of relevant regulatory processes;  
  determine whether a proposed intended use of a substance is currently approved or 
otherwise accepted by FDA; or 
  address general questions about approved food additives, investigational substances, 
FUAs, preparing an FAP, preparing a GRAS notice, or how to legally market a 
substance for use in animal food in the United States.  
 
The following sections provide: (1) best practices for communication between FDA and entities 
engaging in pre-submission consultation, including the development of pre-petition or pre-
notification submissions of a study protocol and evaluation of the adequacy of existing data from 
a completed study, and (2) FDA recommendations to facilitate an entitys development of data 
and information for an FAP or GRAS conclusion.  Such recommendations are intended to help 
ensure that the resulting data and information are useful for evaluating the proposed use of the 
substance in animal food.  A discussion of pre-submission consultation is provided in more detail 
below. 
  6  Contains Nonbinding Recommendations 
 
A.  Pre-Submission Consultation Meetings and General Correspondence Requests 
As stated above, FDA is available to participate in pre-submission consultation meetings 
including in-person meetings, teleconference calls, or video conferences.  After receiving the 
request, FDA will schedule a consultation meeting with you.9  
Alternatively, if you do not intend to participate in a consultation meeting, you may submit 
written questions as general correspondence to the FDA-CVM Director of the Division of 
Animals Feeds.  FDA will provide feedback in a response letter. 
To request a consultation meeting or to send general correspondence inquiries, you may contact 
the Division of Animal Feeds in FDA/CVM in the following ways: 
 
Mail:  Director, Division of Animal Feeds 
Center for Veterinary Medicine 
US Food and Drug Administration 
MPN 4, HFV 220  
12225 Wilkins Avenue  
Rockville, MD 20852 
 
Email:  Animalfood-premarket@fda.hhs.gov 
 
B.  Investigational Food Additive (IFA) File for Animal Food 
 
In addition to pre-submission consultation meetings and general correspondence requests, FDAs 
CVM has a process known as the Investigational Food Additive (IFA) file.  With an IFA file, 
you can exchange confidential data, ideas, and information with FDA using a single 
identification code (IFA file number) while you investigate the safety and utility of a substance 
for its intended use and consider whether to submit an FAP or a GRAS notice.  The IFA file 
process is discussed below. 
 
FDA will open an IFA file and issue you an IFA number, at your request, to provide you the 
opportunity to receive feedback from FDA while you investigate the safety of the intended use of 
a substance in animal food.  You may share data or information with FDA to obtain FDA 
scientific and regulatory expertise and insight.  The types of IFA submissions that FDA reviews 
include: (1) study protocols, (2) FUA requests, and (3) evaluations of the adequacy of existing 
data from completed studies.  
 
An IFA file submission can address one, all, or a combination of issues, such as chemical 
composition, target animal safety, human food safety, and the impact on the environment of the 
proposed use of a substance in animal food.  A submission to an IFA file is not required to 
contain all the elements specified in the regulations for an FAP or a GRAS notice. 
 
                                                 
9 There are additional clearance procedures for non-U.S. citizens requesting an in-person meeting at FDA offices.  
For details, refer to Program Policy and Procedures Manual 1240.2601, https://www.fda.gov/media/70190/download 
or you may email FDA at Animalfood-premarket@fda.hhs.gov.   
 
  7  Contains Nonbinding Recommendations 
 
FDA does not acknowledge the existence of, or announce the establishment of, an IFA file.10  
There are no legally specified time frames for the review of a submission to an IFA file, but FDA 
has created internal goals that are listed throughout this guidance.  FDAs response to an IFA file 
submission is shared only with the entity that established the IFA and with third parties formally 
authorized by the entity. 
 
You may not test market11 a substance until you have reached a GRAS conclusion for the 
intended use, or until a food additive regulation is established for its use in accordance with 21 
CFR part 571; this prohibition includes the time during which FDA is reviewing your IFA file 
submission(s). 
 
While the information submitted to an IFA file will include some or all of the same information 
typically provided later in an FAP or a GRAS notice, the FAP or GRAS notice is independent of 
the IFA file.  You will need to submit all data and information required for an FAP or a GRAS 
notice, even if you have an existing IFA file that contains such information. 
 
If the information to be submitted to an IFA file does not fit into one of the following submission 
types, it can be identified and sent to FDA as general correspondence. 
 
You may submit information such as the following to an IFA file:  
 
1.  Request for Review of a Study Protocol  
 
FDA recommends that entities submit a protocol for a study, not yet initiated, that is intended to 
generate pivotal data to support the utility, safety, stability, or homogeneity of a substance 
intended for use in animal food.  For the purposes of this guidance, pivotal data means data that 
provide the primary basis for a decision or conclusion regarding the proposed use of a substance 
in animal food or the hypothesis to be tested in a study (for example, a hypothesis that a 
substance can be homogeneously mixed into swine feed).  A study protocol may be submitted for 
review during the research and development of a substance.  Additionally, you may include any 
preliminary data generated prior to drafting the study protocol you intend to initiate to provide 
context for the submitted protocol (for example, data from a pilot study or method validation).  
FDAs protocol review can also include an assessment of an analytical method used to generate 
data in support of a proposed use of a substance, and associated method validations.12  FDA has 
set the following goals (from the date of submission receipt) to provide a response letter 
following review of a study protocol review request: 
                                                 
10 Please see section VI. Freedom of Information Act for a discussion of IFA file treatment under the Freedom of 
Information Act.  
11 Test marketing is a tool used to generate insight into the potential market success of a new product, prior to its 
formal launch to a wider customer base.  For an animal nutrition business, this may involve allowing a new product 
to be used in animal food by a select group of customers. 
12 Method validation is the process that helps confirm that the analytical method used is suitable for its intended 
purpose of analyzing a single analyte or a set of analytes. 
  8  Contains Nonbinding Recommendations 
 
  Study protocol without method validation or preliminary data: 50 calendar days from 
the date of submission receipt 
  Study protocol with method validation or preliminary data: 130 calendar days from 
the date of submission receipt 
  Analytical method development and validation: 130 calendar days from the date of 
submission receipt 
 
For review of a study protocol for a study to address the utility, safety, stability, or homogeneity 
of a substance, FDA will primarily be guided by the definition of safety in 21 CFR 570.3(i) and 
information in the current publications of the National Academies of Sciences, Engineering, and 
Medicine-National Research Council (see 21 CFR 570.20).  Additionally, FDA will consider in 
our evaluation principles of experimentation to help ensure that you design and conduct adequate 
and well-controlled studies.  The types of information discussed in Appendix 2 form a general 
framework that FDA uses when evaluating a study protocol based on the above criteria.  FDA 
will not review a protocol for a study that has already been initiated. 
 
When your study is complete, you may submit a final study report (FSR) to FDA as part of a pre-
submission consultation.  You may also use the FSR format to submit data from a completed 
study in an FAP or, as applicable, a GRAS notice.  Please see below for a detailed discussion of 
the FSR in section III.B.3. Request for Review of the Adequacy of Existing Data from a 
Completed Study.  
 
2.  Request for Authorization for Use of Animal Products in Human or Animal 
Foods (FUA Request)  
 
If products, such as meat, milk, or eggs from animals used in studies to support the safety or 
utility of a substance are intended to be used for human or animal food, you must obtain a food 
use authorization (FUA) from FDA or USDA to permit the use of these products (see 21 CFR 
570.17(b)). 
  
FDA will review an FUA request to ensure that the submitted information adequately addresses 
whether food products, such as meat, milk, or eggs, that came from the investigational animals 
that were administered the substance are safe for humans and animals to consume.  If the 
submitted information shows that the consumption of the food products is consistent with public 
health and the food does not contain unsafe levels of residues or metabolites of the 
investigational substance, FDA generally provides an FUA for a specific number of 
investigational animals.  FDA has set the following goal (from the date of submission receipt) to 
provide a response letter following review of an FUA request:  
  FUA request: 130 calendar days from the date of submission receipt 
 
3.  Request for Review of the Adequacy of Existing Data from a Completed Study13  
 
                                                 
13 For purposes of this guidance, study means an experiment, test, or trial. 
  9  Contains Nonbinding Recommendations 
 
You can submit data from completed experiments to receive preliminary feedback from FDA on 
the adequacy and suitability of the data to support relevant sections of an FAP.  FDA may: (1) 
comment on the appropriateness of data or deficiencies present, (2) suggest ways to rectify 
deficiencies, if possible (such as changes to the statistical analysis), and (3) recommend further 
evaluations or additional studies. 
 
FDAs assessment of data will be based principally on whether: (1) the data generated are from 
experiments that were conducted based on accepted scientific methods and principles of 
experimentation, (2) procedures were followed that reflect animal feeding and husbandry 
practices similar to those used in the United States, and (3) nutritional requirements and 
maximum tolerable levels proposed by the National Academies of Sciences, Engineering, and 
Medicine-National Research Council were reasonably met. 
 
When FDA evaluates your submission, data generated from studies conducted in foreign 
countries, meaning countries other than the United States, undergo the same assessment process 
as data generated in the United States.  For the purposes of this guidance, FDA considers foreign 
data to be data generated outside the United States both by entities based within or outside the 
United States.  
 
Based on its preliminary assessment, FDA will provide a written response about the adequacy of 
data you submitted to support the safety of the potential use of a substance in animal food.  In the 
event additional data are required to adequately support such use, FDA will provide relevant 
recommendations, including our scientific rationale for requesting any additional data or 
information.  Unpublished existing data from completed studies can be used to support 
determinations in an FAP.  However, if you intend to use unpublished data to make a GRAS 
conclusion about the use of a substance, you must explain how there could be a basis for a 
conclusion of GRAS status if qualified experts do not have access to such data and information 
(see 21 CFR 570.250(e)).  
 
FDA has set the following goal (from the date of submission receipt) to provide a response letter 
based on the review of existing data from a completed study:  
  Request for review of the adequacy of existing data from a completed study: 180 
calendar days from the date of submission receipt 
 
C.  What to Include in a Request to Establish an IFA File  
 
In addition to providing your name, address, and other contact information, the Agency 
recommends that you provide the following identifying information about the substance and its 
intended use when requesting to establish an IFA file: 
  common or usual name of the substance you intend to investigate; 
  proposed intended use of the substance; 
  target animal species for which you intend to investigate the proposed use of the 
substance; and 
  10  Contains Nonbinding Recommendations 
 
  an Environmental Assessment (EA) or a request for a Categorical Exclusion (CE) 
from preparation of an environmental assessment, including a statement that no 
extraordinary circumstances exist (see section V. Environmental Assessment for more 
details).  
 
If you will be submitting and discussing information with FDA on another entitys behalf, you 
should provide an original letter of authorization signed by the entity.  FDA recommends an 
accurate and complete English translation for any data or information presented in a foreign 
language.14  
 
Once you have received your IFA file number in a letter sent by mail, please refer to it in any 
future correspondence for matters under the scope of that IFA file.  In addition, FDA 
recommends that for each submission type, you provide information to your IFA file as 
described below.  
 
D.  What to Include in Each Submission Type to an Established IFA File 
 
1.  Request for Review of a Study Protocol  
 
The recommended content and format for a study protocol is provided in Appendix 2  Study 
Protocol Content for a Substance Intended for Investigational Use in Animal Food.  The study 
protocol content guideline serves as an outline of information that FDA recommends be included 
to provide the purpose of a study and to explain how a proposed study will be conducted.  The 
items included in the guideline are based on the information you would include in a study report 
or published scientific article that may be used to support an FAP or GRAS conclusion. 
 
While preparing a study protocol, FDA recommends that you follow Appendix 2 as closely as 
possible to provide clear descriptions of the design, materials, methods, steps, and other 
processes involved in the proposed study.  However, not all items may apply to your proposed 
study or you may have additional information to include. 
 
The goals of the Study Protocol Content Guideline in Appendix 2 are to:   
  suggest a uniform format for writing a study protocol;   
  provide a reference of essential items that should be considered for inclusion in a 
study protocol;   
  facilitate the development of a detailed and complete study protocol;  
  help make the protocol more user-friendly;   
  enable FDA reviewers to evaluate the study protocol efficiently and convey their 
comments in terms clearly understood by entities across different geographical 
locations in the world; and 
                                                 
14 While information you provide for the pre-petition consultation process is voluntary, FDAs regulations require an 
accurate and complete English translation of any material submitted in a foreign language as part of an FAP (see 21 
CFR 571.1(a)). 
  11  Contains Nonbinding Recommendations 
 
  reduce the number of significant amendments to a study protocol. 
 
You may use the study protocol format or you may choose to follow alternative procedures to 
design and conduct a study intended to generate data to support regulatory submissions.  
 
2.  Request for Authorization for Use of Animal Products in Human or Animal 
Foods (FUA Request) 
 
You should include in your FUA request data to show that consumption of food derived from 
animals given the maximum levels of the investigational substance with the minimum 
withdrawal periods (if any) will be consistent with the public health and the food will not contain 
unsafe levels of residues or metabolites of the substance.  You also should include the name and 
location of the packing plant where the animals will be processed; however, you may request a 
waiver to exclude the name and location of slaughter.  FDA may place terms on a waiver grant, 
such as an additional holding time prior to slaughter.  
 
You should include the following in your request for an FUA (where applicable):  
  a description of the investigational substance, and its composition, including any 
preservatives, diluents, carriers, or other relevant components; 
  the form in which the substance will be fed (for example, powder, liquid, pellets, 
granules); 
  the maximum use rate that the investigational substance will be administered to 
investigational animals (for example, 300 parts per million in the complete diet, 150 
grams per animal per day, one kilogram per metric ton of diet); 
  information on how the substance will be delivered to the animals for consumption 
(for example, top-dressed on food, mixed into complete diet, added to drinking 
water); 
  the animal species that will receive the substance, including the class (for example, 
growing, adult, breeding) and age; 
  the number of animals that are expected to receive the substance15; 
  a brief description of the study design, including composition of the experimental 
diets;   
  frequency of feeding and timing of substance administration;   
  maximum duration of substance administration; 
  the period for which investigational animals will be held prior to slaughter after 
administration of the substance has stopped (withdrawal period).  A withdrawal 
                                                 
15 The total number of animals you request in your FUA should reflect the actual number of animals you intend to 
treat with the investigational substance in a single study multiplied by the number of times that you plan to repeat 
the study. 
 
  12  Contains Nonbinding Recommendations 
 
period can vary depending on the animal, the substance, and the nature and safety of 
the investigational use of the substance, and could include a zero-day withdrawal16;  
  any other restrictions for using the substance or its conditions of use; and 
  a summary narrative, based on proprietary information and a literature review, as 
applicable, to address the safety of the proposed investigational use of the substance. 
 
3.  Request for Review of the Adequacy of Existing Data from a Completed Study  
 
You may submit data generated from a completed study to FDA for review, preferably in the 
form of an FSR.  The FSR is a complete and comprehensive description of a study, the resulting 
data, the analysis, and conclusions.  The FSR is written after the study is completed to properly 
document and present all relevant details about the study.  An FSR can be used to support an 
FAP.  FDA recommends you use the FSR format in Appendix 3  Investigational Animal Food 
Substance Final Study Report Format: (1) to report data and information generated from a 
completed study, whether generated within or outside (foreign data) the United States, and (2) to 
document data and information you may generate in any future studies, including studies based 
on study protocols previously discussed with FDA. 
 
Using a standardized FSR format will result in more efficient FDA review of your submitted 
data and information that will be a basis of regulatory decisions for an FAP.  The FSR should 
include information from the list provided in Appendix 3 that is applicable to the completed 
study.  The list provided is not exhaustive and is intended to provide a general framework for 
preparing an FSR. 
 
The FSR may be prepared by the study sponsor, the investigator, or both, through a collaborative 
effort.  All individuals or entities involved in the preparation of an FSR should be considered 
authors.  The FSR should include a document signed and dated by the study director (or the 
individual responsible for the overall conduct of the study) describing the contribution provided 
by each author and attest to the accuracy and completeness of the FSR. 
 
Conducting nonclinical laboratory studies to support safety in compliance with the Good 
Laboratory Practice for Nonclinical Laboratory Studies (GLP) regulations helps ensure the 
quality and integrity of the resulting data (see 21 CFR part 58).  FDA recommends that an FSR 
include a statement identifying the parts of the study conducted in accordance with these 
regulations.  If the study or a part of the study was not conducted in accordance with GLP 
regulations, the reasons why should be included in the FSR.17 
 
As noted earlier, unpublished existing data from completed studies can be used to support 
determinations in an FAP.  However, if you intend to use unpublished data to make a GRAS 
                                                 
16 If consistent with public health, a zero-day withdrawal period should be requested for the use of milk from 
investigational animals. 
17 While information you provide regarding nonclinical laboratory studies for the pre-submission consultation 
process is voluntary, FDAs regulations require an FAP to include either a statement that the study was conducted in 
compliance with the requirements set forth in 21 CFR part 58 or a brief statement of the reason for noncompliance if 
the study is not conducted in compliance (see 21 CFR 571.1(k)). 
  13  Contains Nonbinding Recommendations 
 
conclusion about the use of a substance, you must explain how there could be a basis for a 
conclusion of GRAS status if qualified experts do not have access to such data and information 
(see 21 CFR 570.250(e)).  
 
E.  How to Submit Information to an IFA File 
 
To establish an IFA file or add documents to an existing IFA file, FDA recommends you submit 
such information through the CVM eSubmitter Application.  The CVM eSubmitter Application 
is an electronic, question-based submission software to create files that can be transmitted 
electronically to CVM through its secure Electronic Submission Gateway (ESG).  CVMs 
eSubmitter uses data-capturing templates to allow you to assemble a complete and appropriately 
structured submission to CVM.  For detailed explanations on how to submit information through 
the eSubmitter tool, you may reference FDAs website, Electronic Submissions.18  You may 
also reference GFI #108, Registering with CVMs Electronic Submission System.19  
 
Though electronic submission is preferred, you may also submit information by mail to the 
following address: 
 
Director, Division of Animal Feeds 
Center for Veterinary Medicine 
US Food and Drug Administration 
MPN 4, Room 2658  
12225 Wilkins Avenue  
Rockville, MD 20852 
 
Additionally, information may be submitted by email to Animalfood-premarket@fda.hhs.gov.  
 
IV.  FDA Response to Requests Submitted to an IFA File 
 
In response to your request for review of pre-submission consultation information submitted to 
an IFA file, FDA will convey comments from its review in a letter to you.  FDA will strive to 
respond within the number of calendar days indicated earlier in this document; however, FDA 
response time is also dependent on other FDA regulatory priorities.  If review of a study protocol 
without accompanying data is expected to exceed 50 calendar days, FDA will notify you using 
the same method of communication you chose for your study protocol submission. 
 
V.  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 the investigational use 
of a substance.  FDAs regulations in 21 CFR part 25 set forth procedures to supplement the 
NEPA regulations published by the Council on Environmental Quality (CEQ) in 40 CFR parts 
                                                 
18 https://www.fda.gov/animal-veterinary/development-approval-process/electronic-submissions. 
19 https://www.fda.gov/regulatory-information/search-fda-guidance-documents/cvm-gfi-108-registering-cvms-
electronic-submission-system. 
  14  Contains Nonbinding Recommendations 
 
1500-1508.  Under 21 CFR 25.15(a), all applications or petitions requesting Agency action 
require the submission of an Environmental Assessment (EA) or a claim of categorical 
exclusion.  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 substance are both actions that normally require 
preparation of at least an EA, unless the action is of a class that qualifies for categorical 
exclusion. 
 
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 and which have been found to have no such effect in procedures adopted by a 
Federal agency in implementation of the NEPA regulations (see 40 CFR 1508.4).  The classes of 
actions for substances to be used in animal food that typically qualify for a categorical exclusion 
are listed under 21 CFR 25.32.  The establishment of an IFA and actions under that IFA will 
ordinarily qualify for a claim of categorical exclusion under 21 CFR 25.32(b), [a]ction on a 
request for exemption for investigational use of a food additive if the food additive to be shipped 
under the request is intended to be used for clinical studies or research.  
 
To claim a categorical exclusion from the requirement to prepare an EA, you must state in your 
request to establish an IFA that the intended use qualifies for a categorical exclusion, cite the 
particular categorical exclusion that is claimed (e.g., 21 CFR 25.32(b)), and state that, to your 
knowledge, no extraordinary circumstances exist (21 CFR 25.15(a) and (d)).  FDA will review 
the claim of categorical exclusion and determine whether the categorical exclusion is applicable 
and whether any extraordinary circumstances exist that indicate the intended use of the 
investigational substance may significantly affect the quality of the human environment.  
Examples of extraordinary circumstances described in 21 CFR 25.21 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)), and actions that may adversely affect a species 
(flora or fauna) or the critical habitat of a species that has been determined to be endangered or 
threatened under the Endangered Species Act or under the Convention on International Trade in 
Endangered Species of Wild Flora and Fauna, or that is entitled to special protection under some 
other Federal law (21 CFR 25.21(b)).  If you make a claim of categorical exclusion, you may 
consider providing the following language, as outlined in 21 CFR 25.15(a) and (d): 
 
We are requesting a categorical exclusion from preparation of an environmental 
assessment or an environmental impact statement for the investigational use of <insert 
investigational food additive name> as provided in 21 CFR 25.32(b).  To our knowledge, 
no extraordinary circumstances exist that indicate the specific proposed action may 
significantly affect the quality of the human environment. 
 
If extraordinary circumstances are identified, preparation of an EA will be necessary (see 21 
CFR 25.21).  If you think an EA may be needed, FDA recommends that you contact CVM to 
schedule a pre-petition consultation meeting to discuss the requirements of an EA for the 
proposed investigational use of a food additive. 
 
Information concerning the preparation of 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 well-controlled 
  15  Contains Nonbinding Recommendations 
 
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 25.40(e)).  If significant effects requiring the preparation of 
an Environmental Impact Statement (EIS) are identified, then FDA will prepare an EIS.  If such 
effects are not identified, then FDA will prepare a finding of no significant impact (FONSI). 
 
VI.  Freedom of Information Act 
 
Information submitted to FDA is considered a record and is subject to a Freedom of Information 
Act (FOIA) request under 21 CFR part 20.  However, information submitted as part of a pre-
submission consultation to an IFA can be considered information submitted voluntarily to FDA 
relating to an ingredient or product under development.  In accordance with 21 CFR 20.111(d), 
such data and information ordinarily would not be available for public disclosure until referenced 
or presented publicly by you.  If pre-submission information contained in an IFA file is later 
submitted to FDA in an FAP or a GRAS notice, that information would be subject to the 
disclosure provisions under 21 CFR 571.1(h) or 21 CFR 570.275, depending on whether the 
information is included in an FAP or GRAS notice, respectively. 
   
  16  Contains Nonbinding Recommendations 
 
Appendix 1  Table of Abbreviations and Acronyms Used in this Guidance 
 
ABBREVIATION  WHAT IT MEANS 
OR ACRONYM 
CE  Categorical Exclusion  
CEQ  Council on Environmental Quality  
CFR  Code of Federal Regulations 
CVM  Center for Veterinary Medicine 
EA  Environmental Assessment 
EIS  Environmental Impact Statement 
ESG  Electronic Submission Gateway   
FAP  Food Additive Petition 
FDA  U.S. Food and Drug Administration 
FD&C Act  Federal Food, Drug, and Cosmetic Act 
FOIA  Freedom of Information Act  
FONSI  Finding of No Significant Impact 
FR  Federal Register 
FSR  Final Study Report 
FUA  Food Use Authorization 
GLP  Good Laboratory Practice 
GFI  Guidance for Industry 
GRAS  Generally Recognized as Safe 
IFA   Investigational Food Additive  
NEPA  National Environmental Policy Act of 1969 
NRC  National Research Council 
SOP  Standard Operating Procedure 
USC  United States Code  
USDA  Unites States Department of Agriculture 
   
  17  Contains Nonbinding Recommendations 
 
Appendix 2  Study Protocol Content for a Study of a Substance Intended for 
Investigational Use in Animal Food 
 
The following is the general, recommended content of a study protocol for a study of a substance 
intended for investigational use in animal food.  You should make any adjustments needed to 
address the purpose of your study.  
 
1.  TITLE  
 
Provide a descriptive title containing the purpose, substance, and subject of the proposed 
study. 
 
2.  UNIQUE IDENTIFIER FOR THE STUDY 
 
Located on the title page, the unique identifier should include a study protocol number, 
the status of the study protocol (for example, draft, final, amended), and the date of the 
study protocol version.  
 
3.  STUDY LOCATION(S) 
 
Provide the identities and location of study sites, including those where analytical and 
statistical work will be conducted (including contract facilities). 
 
4.  STUDY CONTACTS 
 
Include the investigator(s), sponsor representatives, and all other participants 
responsible for major aspects of the study, such as study monitors, consultants (for 
example, veterinarian, pathologist, or statistician), and quality assurance personnel.  For 
each contact, list the title, qualifications, and summary of professional background (or 
attach a curriculum vitae), as well as the email address, postal address, telephone 
number, and any other methods of communication.  
 
5.  PROTOCOL OBJECTIVE(S) 
 
Define objective(s) of the protocol (for example, determine target animal safety, 
establish utility or functionality). 
 
6.  STUDY OBJECTIVE(S)  
 
Include the following when applicable: 
 
A.  Pivotal/Non-Pivotal Studies - Determine and document whether the study will be 
pivotal or non-pivotal.  Pivotal studies are those used to generate data that provide the 
primary basis for a decision or conclusion regarding the investigational substance and 
the premise that is to be tested in the study (for example, the substance can be 
homogeneously mixed into swine feed, or the substance achieves the intended 
  18  Contains Nonbinding Recommendations 
 
purpose).  Non-pivotal studies are those that are used to generate data that provide a 
secondary or supporting basis for a decision or conclusion regarding the 
investigational substance. 
 
B.  Standards applied - Define any standards that will be applied to the conduct of the 
studies (for example GLP). 
 
C.  Justifications - State, where applicable, information relevant to understanding the 
study objectives, such as data or information from scientific publications or pilot 
studies that justify the conduct of the study. 
 
7.  STUDY SCHEDULE 
 
Include the proposed date(s) of initiation, schedule of events, and proposed date(s) of 
completion. 
 
8.  STUDY DESIGN 
 
Include information, such as treatment groups, experimental design (for example, cross-
over, Latin Square, randomized complete block, split-plots), experimental unit (for 
example, pen, individual animal), blocking factor(s), randomization procedures, such as 
allocation of animals to treatment groups and allocation of treatment groups to 
experimental units, and blinding.  Experimental units should be specified, and its 
selection justified, so that statistical analysis of data is based on the specified 
experimental unit. 
 
9.  STUDY PROCEDURES 
 
A.  Description of Test Animals  Provide descriptions of test animal(s) including: (1) 
age (parity, if applicable), (2) sex, (3) breed and class, (4) initial body weight, (5) 
physiological state (for example, lactating, growing), (6) number of animals, (7) 
source of animals, (8) identification method (animal selection and identification, 
tattoo, neck band, wing band, brand, or other method of identification), and (9) any 
other relevant details. 
 
B.  Inclusion/Exclusion/Post Inclusion Removal Criteria  Provide inclusion, exclusion, 
and post-inclusion removal criteria.  Inclusion criteria is a list of conditions that 
must be met by each study animal (for example, age, body weight, health status) 
prior to placement in the study.  Exclusion criteria is a list of conditions that would 
preclude an animal from being included in the study (for example, illness, 
deformities, stunted growth).  Post-inclusion removal criteria include information 
regarding why an animal would be removed from the study after initial inclusion.  
For post-inclusion removal of animal(s) from the study, provide: (1) qualitative and 
quantitative criteria for removal, (2) procedures for removal of animals from the 
study, and (3) fate of removed study animals. 
 
  19  Contains Nonbinding Recommendations 
 
C.  Acclimation of Test Animals  Provide information regarding the acclimation of 
experimental animals, including: (1) duration, (2) medication and vaccination during 
acclimation period, and (3) the baseline data collected prior to initiating the study, if 
any. 
 
D.  Study Blinding  Describe: (1) the extent of blinding (for example, monitor, 
investigator), (2) the blinding method(s) and procedure(s) used, and (3) the list of 
personnel who will have access to treatment codes. 
 
E.  Analytical Methods  Ensure that details of the analytical methods described in the 
protocol: (1) identify the analytical measurement(s) to be made and explain their 
relevance to protocol objectives, (2) specify the analytical plan to be used, including 
a summary of the method(s), provide a description of procedures for sample 
selection, collection, preparation, and storage, evidence of analytical method 
validations, a description of the validation plan when the method is being developed 
for the study, the quality control procedures for the method, and the criteria and 
procedures used to assess analytical results, (3) reference and summarize relevant 
scientific literature supporting the use of the analytical method for the intended 
measurements, and (4) if applicable, provide documentation that relevant validations 
have been completed before the initiation of the study.  All validation data and 
information should be provided as a separate section. 
 
F.  Study Facilities  Provide information regarding study facilities, including 
containment equipment (cages, pens, runs, stalls, etc.) and space allocation per 
animal (compare to typical United States management practices), lighting equipment 
(if controlled, define photo period), heating and cooling equipment (define 
temperature goals), ventilation equipment, feeding equipment, watering equipment, 
and a facility diagram.  The facility diagram should include: (1) orientation of 
building, (2) location of feeding and watering equipment, (3) placement of 
ventilation system, (4) location of lighting, (5) pen or cage dimensions, (6) 
orientation of experimental blocks, (7) treatment codes, and (8) position of 
environmental and weather recording devices. 
 
G.  Investigational Substance and Control Articles  Provide information regarding the 
investigational substance and control articles.  Identify the investigational substance 
and provide details of its characterization and specifications.  In addition, state the 
control article(s) to be used in the study, if any, and provide its specifications.  
Details, such as chemical name, trade name, lot number, and manufacture and 
expiration dates, of the substance and control articles should be provided.  
Instructions for further mixing (if any), packaging, and storage of these articles 
should be stated.  If administered in drinking water, a complete description of how 
the substance will be incorporated in the water should be provided.  Additionally, 
relevant Safety Data Sheets (SDS) should be provided for the substance and control 
articles.  The specifications of the substance evaluated in investigational studies 
should be similar or identical to the specifications of the substance that will be the 
subject of your FAP or GRAS notice. 
  20  Contains Nonbinding Recommendations 
 
 
H.  Experimental Diet Formulation(s)  Include the following elements in your 
description of the experimental diet formulations: (1) ingredient list(s), (2) nutrient 
and investigational substance concentrations in experimental diet(s), (3) food or 
water sampling procedure(s), (4) food form, and (5) the manufacture of 
experimental diets.  These elements are explained below. 
 
Ingredient lists: describe all ingredients using their common or usual name.  All 
ingredients used in experimental diets should reflect acceptable animal diet 
formulations and feeding practices in the United States. 
 
Nutrient and investigational substance concentration(s) in experimental diet(s): 
include the calculated nutrient concentration (for example, amount of protein, fiber, 
vitamins, minerals), along with the assay procedure(s) (and the analytical laboratory, 
including address and telephone number) that will be used to confirm the accuracy of 
the calculated dietary values.  In addition, provide the quantitative composition of all 
food components or ingredients in the diet (for example, amount of corn, soybean 
meal, wheat). 
 
The food form description: specify the form to be used, such as mashed, crumbled, or 
pelleted. 
 
Manufacture of experimental diets: provide details regarding the procedures, 
facilities, and equipment to be used by the manufacturer of the experimental diets. 
 
Recommendations in reference sources, such as the National Academies of Sciences, 
Engineering, and Medicine-National Research Council publications, may serve as 
useful guides in determining the nutritional requirements of the study animals and the 
preparation of food.  Diet-related study documentation should demonstrate that the 
objective of the study can be accomplished while meeting the animals nutritional and 
associated welfare requirements.  Detailed records of food characteristics should be 
collected in instances where nutritional status can be critical to the measurements to 
be collected in the study. 
 
I.  Experimental Treatment Administration  Provide the feeding program or schedule 
for feeding the experimental diets, including the amount of the substance and how it 
will be administered (for example, mixed in food, top-dressed, mixed in water).  
Provide the frequency and duration of feeding and state whether animals will be 
allowed ad-libitum feeding, limit feeding, or pair-feeding (food amounts matched 
between controls and substance-treated animals). 
 
J.  Concurrent/Concomitant Medications/Therapies/Vaccinations/Fate of Treated 
Animals  List the medications, vaccinations, or other therapies concurrently 
administered to experimental animals.  Describe the fate of animals that are subjected 
to medications during the course of the study and that of animals euthanized due to 
welfare concerns. 
  21  Contains Nonbinding Recommendations 
 
 
K.  General management practices  Include information regarding general 
management practices such as the frequency of personnel visits to the study facility, 
frequency of checking feeders and waterers, frequency of adding food to feeders, 
and frequency of checking for temperature and humidity.  Provide details regarding 
the watering program, the lighting program, and animal welfare checks.  Include the 
facility cleaning schedule, daily monitoring, and any other feeding or management 
program details important to the study. 
 
L.  Provisions for necropsy and disposal of expired test subjects  Describe when, 
where, and who will conduct necropsy examination of animals that die during the 
study and how these animals will be disposed of. 
 
10. SPECIFICATION OF PARAMETERS TO BE ASSESSED 
 
A.  Pivotal parameter(s)  Specify: (1) which parameters will be assessed, (2) when 
treatment responses for each parameter will be assessed, (3) equipment and 
procedures for assessing these responses, (4) forms for capturing source data, (5) 
calculation(s) for derived data, and (6) name(s) and address(es) of outside laboratories 
used for substance or sample analysis. 
 
B.  Supporting parameters  Record: (1) which parameters are supporting parameters, (2) 
when treatment responses for these parameters will be assessed, (3) equipment and 
procedures for assessing these responses, and (4) calculation of derived data. 
 
Units used to measure parameters should be clearly stated and transformation of units or 
conversion factors should be indicated and documented.  When possible, laboratory 
analysis data should be recorded on a record sheet and reference values for the 
parameters used by the laboratory should be presented along with the data.  Source for 
the reference values should be provided, if available. 
 
11. STATISTICAL ANALYSIS OF DATA 
 
Provide a plan for statistical analysis of the data appropriate to the chosen study design 
and the parameters to be evaluated.  Details of the analysis procedure should be 
provided.  When including this information, identify: (1) the experimental unit, (2) the 
number of replicates per treatment and provide details of a power analysis, if applicable, 
(3) the statistical methodology with detailed descriptions of data analysis and treatment 
means separation, hypothesis to be tested or parameters to be estimated, assumptions 
that are being made, and model specification, if a model is to be used, and (4) how the 
statistical results will be used to draw conclusions about the study's objectives. 
 
12. COLLECTION AND RETENTION OF SOURCE DATA 
 
Include information about the following issues: 
 
  22  Contains Nonbinding Recommendations 
 
A.  Data collection  Describe how raw data, either handwritten or electronic, will be 
recorded in a permanent medium (for example, ink for written records or electronic 
records that are unalterable) that can be traced by signature and date to the respective 
individual(s) who recorded the data. 
 
Ensure that data will be collected for all relevant parameters indicated in the study 
protocol.  Source data should be original, accurate, legible, organized, and recorded 
at the time the observation is made.  Authenticate and provide an explanation in the 
record for any data that need to be transcribed or copied for legibility. 
 
Corrections in handwritten study documentation should be made by drawing a 
straight line through the original entry, with the original entry still legible.  Date, 
initial, and provide the reason for the correction.  If data are captured electronically, 
a generally accepted system should be set up to authenticate any changes made to 
such data. 
 
B.  Data Retention  Store all data, information, and other study documentation in a 
manner that protects them from deterioration, destruction, or tampering, in 
accordance with the nature of the records.  The storage site should allow the orderly 
storage and easy retrieval of retained data. 
 
Retain all study documents until after FDA has issued a food additive regulation 
based on the petition or has denied the food additive petition in accordance with 21 
CFR 571.100(a), or until the petitioner has withdrawn the petition, in accordance 
with 21 CFR 571.7.  Although the GRAS final rule does not specify any timeframe 
to retain the data and information that support a conclusion of GRAS status, 
preservation of the data and information that are the basis for the conclusion of 
GRAS status represents prudent practice for those who claim an exclusion from a 
statutory requirement regardless of whether FDA is subsequently notified (81 FR 
54959 at 54992).20 
 
13. AMENDMENT TO THE PROTOCOL/DEVIATION FROM THE 
PROTOCOL 
 
Include how protocol amendments or deviations will be addressed during the conduct of 
the study. 
 
A.  Amendments  The investigator and the study site(s) personnel (if applicable) should 
document, sign, and date any amendments made to the protocol.  Protocol 
amendments should include a justification.  A copy of the signed protocol and any 
amendments should be maintained at the study site. 
 
                                                 
20 https://www.federalregister.gov/documents/2016/08/17/2016-19164/substances-generally-recognized-as-safe.  
  23  Contains Nonbinding Recommendations 
 
B.  Deviations  The investigator and study site(s) personnel should record and promptly 
report deviations to the study sponsor.  Deviations should be authorized by the 
sponsor in writing. 
 
14. INVESTIGATIONAL SUBSTANCE DISPOSITION/ANIMAL 
ACCOUNTABILITY/FOOD DISPOSITION/FOOD ACCOUNTABILITY 
 
Document the accountability of all investigational substances used and the disposition 
of all unused investigational substances, unused food, and animals used.  You must 
obtain food use authorizations, if products from animals that consume the substance are 
intended for use as human or animal food. (21 CFR 570.17(b)).  Food use authorization 
should be obtained prior to conducting the study (see section III.D.2. Request for 
Authorization for Use of Animal Products in Human or Animal Foods (FUA Request) 
above). 
 
15. PROTOCOL APPROVAL SIGNATURES 
 
Obtain dated signatures from the study leadership (for example, project manager, study 
sponsor, investigator, study monitor, quality assurance representative, consultant) on the 
protocol, including any amendments to the protocol, to indicate approval of the protocol 
and agreement to follow it. 
 
16. APPENDICES 
 
Include relevant appendices, such as a description of sample collection(s), description of 
sample preparation, description of special equipment, analytical methods, and validation 
techniques employed.   We also recommend including standard operating procedures 
(SOPs), in order for us to assess whether a study was well conducted and thus suitable 
to serve as the basis for a future regulatory decision. 
  
   
  24  Contains Nonbinding Recommendations 
 
Appendix 3  Investigational Animal Food Substance Final Study Report Format 
  
The following is the general, recommended content of a final study report (FSR) for an 
investigational substance intended for use in animal food.  You should make any necessary 
adjustments needed to address the purpose of your particular report. 
 
1.  Title of the study 
 
Provide a descriptive title containing the purpose, investigational substance, and subjects 
of the study. 
 
2.  Unique Identifier for the study report 
 
Located on the title page, the unique identifier should include a study report number, the 
status of the study report (such as, final or amended), and the date of the version of the 
study report. 
 
3.  Study Objectives 
 
State the purpose, type of study, name of the substance evaluated, target animal, and any 
other information relevant to the study objectives. 
 
4.  Study personnel 
 
Provide the title or roles played, qualifications, summary of professional background (or 
attach a curriculum vitae), and contact information including postal address, telephone 
number, and email address for all study personnel.  Study personnel may include the 
investigator(s), representatives of the sponsor, study monitor(s), quality assurance and 
quality control personnel, consulting veterinarian(s), pathologist(s), statistician(s), and all 
other participants in the study. 
 
5.  Study location(s) 
 
Provide the identities and location of study sites, including those where analytical and 
statistical work were conducted (including contract facilities). 
 
6.  Key study dates 
 
Include key study dates such as the start and finish dates for the study, dates of sample 
collection and analyses, dates of study termination, database lock date, and dates of 
statistical processing of data. 
 
7.  Study design 
 
Include the type and description of study design (for example, cross-over, Latin Square, 
randomized complete block, split-plots).  Treatments, replicates, blocking factors, 
  25  Contains Nonbinding Recommendations 
 
randomization procedures, and any other design components should be addressed.  The 
experimental unit should be specified, and its selection justified, so that statistical 
analysis of data is based on that specified experimental unit. 
 
8.  Materials and methods 
 
Include details of materials and methods of the study, such as animal selection and 
identification, characterizations and compositions of the substance and control article(s), 
treatment diets used, analyzed dietary nutrient compositions, treatment blinding to study 
personnel, study animal selection (including details of inclusion, exclusion, and removal 
criteria), duration of acclimation, treatment and observation periods, removal of study 
animals during the course of the study, necropsy details, disposal of investigational 
substance, and any other substances, resources, approaches or procedures of the study.  
Materials and methods also include details of data collection, in addition to sample 
collection, processing, handling, and storage.  Description of the analytical methods used 
in the study, including any applicable assay methods used to determine the 
investigational substance or relevant metabolites in food, water, body fluids, or tissues, 
should be included. 
 
9.  Housing and animal management 
 
Include details of how the animals were housed and maintained during the study.  This 
includes details of animal housing (for example, cage or litter for broiler chickens, pens 
or pasture for cattle), feeding, watering, ventilation, heating and cooling, lighting 
schedules, welfare checks, food addition, food withdrawal, animal weighing, sample 
collection, euthanasia of animals, and any other details about how the animals were 
housed, maintained, and handled during the study. 
 
10. Statistical methods 
 
Describe the statistical methods used to analyze the data generated from the study.  
Information such as hypotheses tested, parameters estimated, assumptions made, level of 
significance, experimental unit, and statistical model should be addressed.  The number 
of animals used in the study should be justified in terms of the power of the study and 
pertinent considerations relevant to standard animal husbandry practices in the United 
States.  Descriptions of the transformations (if any), calculations, or operations performed 
on the raw data should be provided, and the specific statistical methods used to analyze 
the raw data and separation of treatment means should be explained.  Appropriate 
justification should be provided if any data points are excluded from analyses and reasons 
should be given if the statistical methods used differed from those initially planned in the 
studys protocol.  Removal of data points based on numerical outlier analyses alone is not 
recommended and may impact determinations made from the review of a study report. 
 
11. Results 
 
  26  Contains Nonbinding Recommendations 
 
Describe the results of the study, whether favorable or unfavorable, including summary 
tables and diagrams of all relevant data recorded during the study.  Each data table or 
summary diagram should be labeled so that the reader can understand the information 
provided in the data table or summary diagram.  Summary results presented should show 
the treatment means using appropriate mean separation notations, following statistical 
analysis and mean separation. 
 
12. Discussion 
 
Include a discussion section, which is primarily used to interpret and explain study 
results, answer the question(s) for which the study was conducted, justify the study 
methods, and to critically and objectively evaluate the study results.  In a narrative 
format, provide your interpretation of the study results and conclusions based on each of 
the treatment effects, interactions if any, or other observations in relation to the objective 
of the study. 
 
13. Relevant administrative and compliance items 
 
Provide a summary description of the procedures used to record, process, handle, and 
retain samples, raw data, and other study documentation.  This includes descriptions of 
any protocol deviations and amendments, and an assessment of their impact on the 
outcome of the study, in addition to a description of circumstances that could have 
affected the quality or integrity of the data, specifying the time frame and the extent of 
their occurrence.  Information on the occurrence of adverse events or lack of thereof, and 
any measures taken in response to an adverse event should be described in detail. 
 
14. Relevant additional information 
 
Include information such as amended study protocols, dates of monitor visits, audit 
certification, supplementary reports (such as statistical and analytical reports), copies of 
standard operating procedures (SOPs) or a summary narrative of information in the 
SOPs, or any other relevant additional information.  Any additions, deletions, or 
corrections to the FSR should be included as an amendment, signed and dated by the FSR 
author(s).  
 
  27