GUIDELINES ON USE OF PHARMACEUTICAL-/NON-PHARMACEUTICALGRADE SUBSTANCES ON ANIMALS
A pharmaceutical-grade substance is defined as any active or inactive drug, biologic or reagent,
for which a chemical purity standard has been established by a recognized national or regional
pharmacopeia (e.g., the U.S. Pharmacopeia (USP), British Pharmacopeia (BP), National
Formulary (NF), European Pharmacopoeia (EP), Japanese Pharmacopeia (JP), etc.). These
standards are used by manufacturers to help ensure the products are of the appropriate
chemical purity and quality, in the appropriate solution or compound, to ensure stability, safety,
and efficacy.
Whenever possible, pharmaceutical-grade substances must be used for the clinical treatment of
animals and to prevent or reduce/eliminate animal pain or distress, even in acute procedures.
Non-pharmaceutical grade substances should only be used in clinical treatment after specific
review and approval by the IACUC for reasons such as scientific necessity or non-availability of
an acceptable veterinary or human pharmaceutical-grade product. Cost savings alone is not an
adequate justification for using non-pharmaceutical-grade substances in clinical treatment.
For research purposes, i.e when the compounds are used to accomplish the scientific aims of
the study, pharmaceutical-grade substances are preferred, if available, and suitable. If the use
of non-pharmaceutical-grade substances is essential for the conduct of science, the goal of the
IACUC is to consider the health and well-being of the animals while aiding the researcher in
minimizing potentially confounding experimental variables and maximizing reproducibility of the
research.
(A) Use of pharmaceutical-grade substances
(a) Pharmaceutical-grade substances should be reconstituted according to the product
insert.
(b) Any excipient used in the formulation such as vehicle, diluent and others not described in
the product insert should be of pharmaceutical-grade. Commonly used solvent / vehicle
such as normal saline, Phosphate Buffered Saline (PBS) and Dimethyl sulfoxide
(DMSO) are available in pharmaceutical-grade. Pharmaceutical-grade substances
adulterated by non-pharmaceutical-grade excipients will render the preparation nonpharmaceutical-grade.
(c) Reconstitution or formulation should be done under aseptic conditions. Sterilized pipette
or pipettor tips should be used in the transfer of reconstitutents or vehicles.
(B) Use of non- pharmaceutical- grade substances
The use of non-pharmaceutical-grade compounds in experimental animals will only be
acceptable by IACUC in the following situations.
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Use is compliant with applicable national regulatory guidelines and requirements
and the requirements of relevant funding agencies
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The pharmaceutical-grade compound is not available in the appropriate
concentration or formulation or the appropriate vehicle control is unavailable in
pharmaceutical-grade.
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The compound is required to generate data that are part of an ongoing study or that
are comparable to previous work
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The chemical properties of the compound are appropriate for the study and the route
of administration (e.g., the purity, grade, stability in and out of solution, solution
vehicle properties, pH, osmolality, and compatibility of the solvent and other
components of final preparation). In some cases the reagent-grade of the compound
may be as or more pure than the pharmaceutical-grade; and their use is well
documented in scientific literature.
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The method of preparation, labeling (i.e., preparation and use-by dates),
administration and storage of formulations is appropriately considered with the aim of
maintaining their stability and quality (i.e., to prevent inadvertent co-administration of
infectious agents or contaminants).
(C) Consideration of factors affecting the health and well-being of animals
When (i) non-pharmaceutical-grade substances, (ii) pharmaceutical- grade substances
adulterated with non-pharmaceutical-grade excipients, and (iii) pharmaceutical- grade
substances not appropriate for the species and route(s) of administration are used, the
following factors affecting the health and well-being of animals should be considered and
documented in the animal use protocol:
1. Vehicle, diluent and solvent
Consideration should be given to formulations using pharmaceutical-grade excipient
such as vehicle, diluent and others. When non-pharmaceutical -grade excipient is
used, it should be sterile and physiologic.
2. Grade and purity.
The highest-grade equivalent chemical reagent should be used and formulated
aseptically and with a non-toxic vehicle as appropriate for the route of administration.
3. Sterility
All formulations must be prepared under strict aseptic conditions and/or sterilized
prior to use. A 0.22 um filter could be used to filter-sterilize heat sensitive
formulations.
4. pH
The pH range of the internal environment of animal is very narrow (e.g., limited of 7.4
± 0.4 in human). Formulation with high or low pH should be adjusted or diluted with a
buffer if possible. Formulations of high or low pH given by the intra-muscular or
subcutaneous will cause irritation, pain or tissue necrosis. The pH for these routes
should be 7.3 to 7.45 or alternative routes such as intra-peritoneal or intra-venous
should be considered. The pH of the formulation for IV or IP should be between 4.5
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and 8.0. Use of formulation outside this pH range if required, must be administered in
small volume. The effect of pH on the solubility of the substance should also be
considered as precipitation may occur when mixed with blood, resulting in vascular
occlusion, emboli, and thrombosis of local and distant capillary beds.
5. Osmolality
The formulation should be isotonic or adjusted if it is not, whenever possible. Nonisotonic formulations may either cause red blood cell crenation or haemolysis when
administered via the intravenous route, or may cause localized tissue damage and
associated pain upon injection. Non-isotonic formulations should be administered
slowly and in small volume.
6. Stability
The stability of the formulation should be considered to ensure that the dose is
effective and no harmful degradation by- products may compromise the experimental
results or the welfare of animals based on available information. Methods of storage,
such as dark bottles for light-sensitive compounds, refrigeration or freezing for
compounds unstable at room temperature, etc. should be determined. Formulations
should be used as soon as possible, or within a week if the shelf-life of the
preparation is unknown and depending on the usage of the compound i.e. possible
contamination after multiple use from the same vial. Use of fresh preparation should
be considered if any unstable compound is included in the formulation. The
preparation should be labeled with the name, concentrations, date of preparation and
expiry date of the formulation, and should be discarded if changes in color and/or
precipitation occur.
7. Pharmacokinetics
Information on the pharmacokinetic of the substance should be noted where
available. Such information will assists in determination of the dose, route and
frequency and efficacy of the substance.
8. Toxicity, pyrogenicity, compatibility and side effects
The toxicity, pyrogenicity, compatibility and side effects available in the literature
should be reviewed. These factors cause pain and distress to animals postadministration and measure to be taken to minimize pain and distress should be
documented. Distinction should be made between the intended effects and potential
side effects. Toxic substances should be used at lowest possible effective doses for
the intended effect and at shortest possible humane endpoints. Pyrogens, such as
endotoxins, may cause fever when injected into an animal. Purity and sterility of a
non-pharmaceutical-grade substance do not assure that pyrogens are not present.
Pyrogenicity is a potential experimental variable that PI should be aware of when
using non-pharmaceutical grade drugs. Pyrogens can be avoided by using purified
and characterized compounds and diluents in preparation of the final formulation.
Substance that is not compatible may cause tissue damage when administered to
animals. The above and other side effects in animals should be monitored daily.
9. Pilot Study
An efficient way to reduce the negative impacts on laboratory animals is the use of a
pilot study. If a novel compound is being tested, a pilot study using a minimum
number of animals should be conducted to determine the dose and induced adverse
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effects. This would in turn help determine humane end point criteria specific to the
project.
References.
The Guide for the Care and Use of Laboratory Animals. NRC. ILAR. Eighth Ed. 2011. P.31.
http://grants.nih.gov/grants/olaw/Guide-for-the-Care-and-Use-of-Laboratory-Animals.pdf
Administration of Substances to Laboratory Animals: Equipment Considerations, Vehicle
Selection, and solute Preparation. Journal of the American Association for Laboratory Animal
Science. September 2011, Vol 50, No 5, 614–627
Administration of Substances to Laboratory Animals: Routes of Administration and Factors to
Consider. Journal of the American Association for Laboratory Animal Science. September 2011
Vol 50, No 5 Pages 600–613
A good practice guide to the administration of substances and removal of blood, including
routes and volumes. Karl-Heinz Diehl, Robin Hull, David Morton, Rudolf Pfister, Yvon
Rabemampianina, David Smith, Jean-Marc Vidal, Cor Van De Vorstenbosch . J Appl Toxicol 21
15-23, 2001.
Refining procedures for the administration of substances. Report of the BVAAWF/ FRAME/
RSPCA/ UFAW joint working group on refinement. D.B. Mortom, M. Jennings, A. Beckwell, R.
Ewbank, C. Godfrey, B. Holgayte, I. Inglis, R. James, C. Page, I. Sharman, R. Verschoyle, L.
Westall and A.B. Wilson. Laboratory Animals 35, 1-41, 2001.
Office of Laboratory Animal Welfare: Educational Resources
http://grants.nih.gov/grants/olaw/educational_resources.htm
Recognition and Alleviation of pain in Laboratory Animals. Institute of Laboratory Animal
Research, National Research council 2009 P 124
AAALAC “Frequently Asked Question” on the use address non-pharmaceutical-grade
compounds.
www.aaalac.org/accreditation/faq_landing.cfm
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