An innovative antinociceptive test, the cold ethanol tail-flick test (CET), was developed for evaluating the actions of opioid analgesics. To select an optimal operation temperature range for the CET, temperatures from -5 degrees C to -30 degrees C were screened. After screening, temperatures ranging between -20 degrees C and -30 degrees C were both strong and effective enough to act as a noxious cold stimulus. In the following study, -20 degrees C was selected as the cold stimulus for the CET. The sensitivity and specificity of this test were challenged by opioid analgesics: an agonist (morphine) and two agonist-antagonists (buprenorphine and nalbuphine), two tranquilizers (droperidol and diazepam), and four nonopioid analgesics (acetaminophen, aspirin, indomethacin, and ketoprofen). The sensitivity of the CET was also compared with the assays using heat (radiant heat and hot water). The AD50 values determined by the CET for morphine, buprenorphine, and nalbuphine were 0.16 mg/kg, 0.22 micrograms/kg, and 0.19 mg/kg, respectively. Naloxone, an opioid antagonist, blocked the antinociceptive effects of these opioids which were determined by the CET. Furthermore, the tranquilizers and nonopioid analgesics did not show any activity in the CET. Our results show that not only can the CET assess the antinociceptive activity of both opioid agonist and mixed agonist-antagonist, it also possess the characteristics of sensitivity, specificity, simplicity, and reproducibility.