H.1.2.

4 Hot plate method

PURPOSE AND RATIONALE

The paws of mice and rats are very sensitive to heat at temperatures which are not damaging the skin. The responses are jumping, withdrawal of the paws and licking of the paws. The time until these responses occur is prolonged after administration of centrally acting analgesics, whereas peripheral analgesics of the acetylsalicylic acid or phenyl-acetic acid type do not generally affect these responses.
PROCEDURE

ate agonists-antagonists provide unreliable results. The validity of the test has been shown even in the presence of substantial impairment of motor performance (Plummer et al 1991). Mixed opiate agonistsantagonists can be evaluated if the temperature of the hot plate is lowered to 49.5C (OCallaghan and Holtzman 1975, Zimer et al 1986).
MODIFICATIONS OF THE METHOD

ONeill et al (1983) described an automated, highcapacity method for measuring jump latencies on a hot plate. A hot-plate test with increasing temperature was recommended by Tjlsen et al (1991).
REFERENCES
Eddy NB, Leimbach D (1953) Synthetic analgesics: II. Dithienylbutenyl- and dithienylbutylamines. J Pharmacol Exp Ther 107:385393 Jaco J, Blozovski M (1961) Action des divers analgsiques sur le comportement de souris exposes a un stimulus thermoalgsique. Arch Int Pharmacodyn 138:296309 Jacob J, Loiseau G, Echinard-Garin P, Barthelemy C, Lafille C (1964) Caractrisation et dtection pharmacologiques des substances hallucinognes. II.-Antagonismes vis-a-vis de la morphine chez la souris. Arch Int Pharmacodyn 148:1430 Kitchen I, Crowder M (1985) Assessment of the hot-plate antinociceptive test in mice. A new method for the statistical treatment of graded data. J Pharmacol Meth 13:17 Knoll J (1967) Screening and grouping of psychopharmacological agents. In: Siegler PE, Moyer HJ (eds) Animal and Clinical Pharmacologic Techniques in Drug Evaluation. Yearbook Med Publ. Inc., Chicago, pp 305321 ONeill KA, Courtney C, Rankin R, Weissman A (1983) An automated, high-capacity method for measuring jump latencies on a hot plate. J Pharmacol Meth 10:1318 OCallaghan JP, Holtzman SG (1975) Quantification of the analgesic activity of the narcotic antagonists by a modified hot plate procedure. J Pharm Exp Ther 192:497505 Plummer JL, Cmielewski PL, Gourlay GK, Owen H, Cousins MJ (1991) Assessment of antinociceptive drug effects in the presence of impaired motor performance. J Pharmacol Meth 26:7987 Tjlsen A, Rosland JH, Berge OG, Hole K (1991) The increasing temperature hot-plate test: an improved test of nociception in mice and rats. J Pharmacol Meth 25:241250 Witkin LB, Heubner CF, Galgi F, OKeefe E, Spitaletta P, Plummer AJ (1961) Pharmacology of 2-aminino-indane hydrochloride (SU 8629): a potent non-narcotic analgesic. J Pharmacol Exp Ther 133:400408 Woolfe G, MacDonald AD (1944) The evaluation of the analgesic action of pethidine hydrochloride (DEMEROL) J Pharmacol Exper Ther 80:300307 Zimer PO, Wynn RL, Ford RD, Rudo FG (1986) Effect of hot plate temperature on the antinociceptive activity of mixed opioid agonist antagonist compounds. Drug Dev Res 7:277280

The method originally described by Woolfe and Mac Donald (1944) has been modified by several investigators. The following modification has been proven to be suitable: Groups of 10 mice of either sex with an initial weight of 18 to 22 g are used for each dose. The hot plate, which is commercially available, consists of a electrically heated surface. The temperature is controlled for 55 to 56C. This can be a copper plate or a heated glass surface. The animals are placed on the hot plate and the time until either licking or jumping occurs is recorded by a stop-watch. The latency is recorded before and after 20, 60 and 90 minutes following oral or subcutaneous administration of the standard or the test compound.
EVALUATION

The prolongation of the latency times comparing the values before and after administration of the test compounds or the values of the control with the experimental groups can be used for statistical comparison using the t-test. Alternatively, the values which exceed the value before administration for 50% or 100% can be regarded as positive and ED50 values can be calculated. Doses of 7.5 mg/kg s.c. morphine hydrochloride, 30 mg/kg s.c. codeine hydrochloride, 30 mg/kg s.c. pethidine hydrochloride and 400 mg/kg s.c. phenazone were found to be effective, whereas aspirin showed no effect even at high doses.
CRITICAL ASSESSMENT OF THE TEST

The hot plate test has been used by many investigators and has been found to be suitable for evaluation of centrally but not of peripherally acting analgesics. Mice as well as rats have been used. The method has the drawback that sedatives and muscle relaxants (Woolfe and MacDonald 1944) or psychotomimetics (Knoll, 1967) cause false positives, while mixed opi-