This was performed by the hotplate
test, where a rat is placed on a heated plate,
and if the rat is not injected with analgesic,
it will jump up from the pain.
Even on injection of 5 microgram
(1mcg=1 millionth of a gram) of epibatidine, the
rat would not jump, proving that epibatidine was
200 times more effective than morphine. Even when
injected with naloxone, a drug negating the effect
of morphine, the rat would not jump, showing epibatidine
operates on a different pathway from opioids.
Tragically, 750 frogs had to
be killed to produce just 60mg for analysis, so that
with this one off sacrifice, artificial synthetic routes
could be devised. Even so, at the time, without the
aid of spectroscopic methods, the structure could not
Equador did not allow more frogs to
be taken from the rainforest, and as such small amounts
were extracted from each frog, (a tenth of a milligram),
an attempt was made to breed the frogs in captivity.
This failed, as it was discovered that the captive frogs
produced no epibatidine, leading the researchers to
conclude that the alkaloid was derived from the frog's
diet and environment. With no new source of epibatidine,
and the remaining amount insufficient for further research,
the sample was stored away for over a decade.
Continue on to cigarettes...