A series of three practical sessions
are designed to give students
firsthand experience with the preparation of natural product extracts
and assay using a live tissue preparation. Areca or betel nuts are
the seeds from the fruit of the Areca catechu palm
tree that is known to contain a number of pharmacologically active
alkaloids. The principal of these is arecoline that makes up to 1%
of the dry nut. Arecoline is a potent spasmogenic agent, causing smooth
muscle contraction via muscarinic acetylcholine receptor (mAChR) activation.
The first session involves the preparation of methanolic extracts
from whole areca nuts and TLC for the qualitative identification of
arecoline present in the extract. The second session utilizes the
spasmodic effects of arecoline on smooth muscle to allow students
to perform a live tissue bioassay using guinea-pig ileum. This response
is subsequently blocked by the mAChR antagonist atropine to investigate
the mechanism underlying the measured response. The final session
gives students the opportunity to construct arecoline dose–response
curves based on their experimentally derived data. From this curve
and the obtained antagonist results, they are able to calculate an
estimate of the arecoline content in the extracts they prepared and
the original betel nut samples.
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