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Non Pharmaceutical Grade Pentobarbital Use19

non-pharmaceutical-grade-pentobarbital-use19

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19 views3 pages

Non Pharmaceutical Grade Pentobarbital Use19

non-pharmaceutical-grade-pentobarbital-use19

Uploaded by

dre kings
Copyright
© © All Rights Reserved
We take content rights seriously. If you suspect this is your content, claim it here.
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GUIDELINES FOR THE PREPARATION AND USE OF NON-

PHARMACEUTICAL GRADE PENTOBARBITAL IN LABORATORY ANIMALS

The guidelines presented here are extracted from documents describing the requirements
and expectations for the use of non-pharmaceutical grade compounds, with references to
the regulations, policies, and guidelines of the major animal research oversight
organizations.1-4

A pharmaceutical-grade compound (PGC) 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)). These
standards are used by manufacturers to help ensure the products are of the appropriate
chemical purity and quality, and in the appropriate solution or compound, to ensure
stability, safety, and efficacy.2 The Food and Drug Administration (FDA) maintains a
database listing of FDA approved commercial formulations for both FDA approved human
drugs (the Orange Book) and veterinary drugs (the Green Book).

According to regulations regarding animal care and use, investigators MUST use
pharmaceutical grade compounds whenever those are commercially available. However,
because of the high cost and limited availability of Nembutal (pentobarbital 50 mg/ml
injection), the Office of Laboratory Animal Welfare (OLAW) has placed it into the
“unavailable category” 4, meaning that nonpharmaceutical grade products can be used
instead.

Pentobarbital from a reagent or analytical-grade powder, properly prepared with assurance


of sterility, appropriate storage and handling, and approval by the IACUC is acceptable.
Cost saving alone is not a valid reason for use of a non-pharmaceutical grade compound
(non-PGC), and that its use must be justified in your protocol and approved by the IACUC.

The use of a non-PGC pentobarbital solution must meet the same standards as for use in
any other application.3 These guidelines pertain to all components, both active and
inactive, contained in the preparation to be administered. Therefore, the vehicle used to
facilitate administration of a compound is just as important of a consideration as the active
compound in the preparation. Veterinary drugs that are reconstituted in a manner not in
accord with these guidelines will not be considered acceptable.

Preparation of Pentobarbital Solutions: 5


When drugs or chemicals are formulated for injection, they must be prepared in a sterile
manner. Sterile constituents (e.g. sterile drug or chemical, sterile diluents, etc.), a sterile
container, and a means of keeping the preparation sterile are required. Sterile injection
vials are preferred as they make it easier to load a syringe and allow removal of solution
without exposing the contents to contaminants. All handling of pentobarbital powders must
be done in accordance with the manufacturer’s/supplier’s safety data sheet (SDS).

Updated 11-5-19
Diluents or vehicles must be sterile acceptable components. Exceptions must be included
in the protocol in order to be evaluated by the IACUC on a case-by-case basis.
Containers must be labeled with the drug name, concentration, date of preparation, and the
date of expiration. Prepared solutions must be passed through a sterile syringe filter (0.22µ
or finer) at the time of preparation. This can be done in the process of transfer to an
injection vial. If there is any question about the sterility of a prepared or stored solution, it
must be discarded.

Pentobarbital solutions prepared and stored properly in a suitable injection vial must be
used within 7 days of preparation. Therefore it is prudent to prepare only as much as can
be used in a reasonable period of time. Drugs must be secured properly in a locked cabinet
and records kept. Solutions must not be used if they are cloudy, discolored, precipitated,
or have become otherwise altered in appearance since initial preparation. Any solution
remaining after 7 days must be disposed of through proper channels. Expired drug
solutions in containers must be labeled (e.g., “expired—Dispose—do NOT use in
animals”) and stored separately from drugs in-date. Since pentobarbital is a DEA
controlled substance, it must used, stored and discarded in a manner consistent with
Federal, State and RFUMS policies.

Pyrogenicity is a potential experimental variable that researchers should be aware of when


using non-pharmaceutical grade drugs. Pyrogens, such as endotoxins, may cause fever
when injected into an animal. Sterility by filtering does not ensure that pyrogens are not
present. The core temperature of animals anesthetized with non-USP pentobarbital sodium
solution should be monitored for potential fever.

Recipe for 100 ml pentobarbital sodium:

INGREDIENTS
• 6 gm pentobarbital sodium USP
(Reagent or analytical grade may be substituted if USP is unavailable but purity
must be > 97%)
• 10 ml ethanol (95%) USP
• 40 ml propylene glycol USP
• Water for injection USP qs 100 ml
• Sodium hydroxide USP or hydrochloric acid USP as required bringing pH to ~ 9.5

MIXING
The mixing process must be performed in a certified Class II biosafety cabinet. Masks,
gloves, and lab coats are to be worn during preparation, and MSDS procedures for safe
handling followed.

• Dissolve the pentobarbital powder in ethanol.


• Add 25 ml of water for injection (but only after the pentobarbital is completely
dissolved); mix thoroughly.
• Add 40 ml propylene glycol; mix.
• Bring to final volume (100 ml) with water for injection. Adjust pH as required.

Updated 11-5-19 2
• Sterilize by filtration in the biosafety cabinet into a sterile empty vial for injection.
• The pentobarbital sodium concentration in the final solution is 60 mg/ml.
• Recipe may be proportionally adjusted to produce smaller quantities of final product

NOTES
As for every controlled substance, PI MUST have valid Federal and State controlled
substance licenses and all use of these agents must be recorded as a Schedule II
controlled substance. The PI retains all responsibility for purchase, storage, and use of
controlled substances within his/her laboratory. By publishing these guidelines,
RFUMS and /or the IACUC do not assume any of this responsibility.

References:
1
U.S. Department of Agriculture, Animal and Plant Health Inspection Service, Animal
Care, Policy 3-Veterinary Care, March 25, 2011.
2
AAALAC frequently asked questions about Non-Pharmaceutical Grade Compounds
3
OLAW Position Statement- Non-Pharmaceutical Grade Substances
4
Transcript of OLAW On-line Seminar broadcast on March 1, 2012 - Use of Non-
Pharmaceutical-Grade Chemicals and Other Substances in Research with Animals
5
www.iacuc.ucsf.edu/Policies/non-pharmaceutical

Updated 11-5-19 3

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