Conventional Test Method:

Mouse LD50 Assay for Botulinum Toxin Potency Testing

Alternative Test Method:

Name & Description:

Mouse Phrenic Nerve (MPN) Assay

A phrenic nerve preparation is isolated from the mouse diaphragm and placed in Krebs-Ringer solution where the resting tension of the diaphragm is measured. The incubation medium is then exchanged for toxin containing medium and the diaphragm tension is monitored until there is a 50% decrease in amplitude. This information is used to construct a dose-response curve.

Validation Status:

US:  ICCVAM workshop panel recommended that further validation studies be conducted (2008)

Germany: Validated by the Medical School of Hanover

Regulatory Status:

EU: Listed in the European Pharmacopoeia (2005)

Effect or Potential Effect on Animal Use:

Reduction
(At least 50% decrease in numbers of animals used)

Refinement
(Uses killed instead of live mice)

References:
Information Last Reviewed:
May 2012