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Reference No. C4.1
Research Conference, Canada - August, 1982
I T. L. Biological Pro a
M.S. Bi I imorl a
S.R. Massey
At the 1981 Research Conference it was acreed that there was a need
for an increased understanding of the effect of Product changes on biological
activity and the related chemical composition/biological activity interface.
Although these areas have been studied extensively over the years,
only limited progress was made due, in part, to the type of test procedures
used. However, the utility of the Ames test has now been recognised and
represents a valuable tool in the study of the biological acti vi ty of tobacco
smcke.
The I T. L. contribution to tne Group Biological Programme is as follows:-
1. In-House Studies
Mutagenicity studies to investioate the contribution of a range of
factors on Ames biological activity. Tobacco smoke condensate will be
subjected to limited fractionation Procedures and the biological activity of each
fraction and of whole smoke determined. The factors that will be considered are:-
(i) Smoking re i.e., human versus standard machine smoking.
(ii) Different tobacco types including reconstituted materials.
(iii) Product design.
By analysis of the matrix of results it should be possible to determine
whether or not there are any consistent activity/fraction patterns. Subsequently
an attempt will be made to characterize biologically active fractions. However,
this work would, of necessity, have to be carried out at Southampton since the
necessary analytical techniques are only available there. This implies a
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commitment from GR & DC towards this project, although the analytical work
would probably not commence unti I mid 1983; probably the HPLC expertise
available within the biological sciences group would be used to separate
components of active fractions, with a view to analysis by aopropriate
analytical procedures such as FTIR, MS etc.
Such a study will complement the deproteinization programme proposed
within the group which will be carried out by GR & DC. Earlier work in
ITL has shown that Ames ,mu is highly correlated with both the
protein and total nitrogen content of tobaccos.
It is desirable that contact be made with the group's mu tagen
consultant, Prof. B.A. Bridges, regarding information an the fractionation
of motor car exhaust condensate, an area in which he is known to be active.
eon',,, This may be relevant since nitrogen containing compounds of high activity
are reputedly present in such condensates. The project also implies close
collaboration between Montreal and Southampton and on-site discussions
between the participating scientists will be required. Dr. Bilimoria will be
conducting a literature review before starting work, since there are some
published studies in this area.
In addition, the study will include an examination, including
fractionation, of sidestream smoke condensate. The fact that sidestream
smoke appears to have similar activity to mainstream smoke (but very different
chemical composition) merits further investigation. Sidestream smoke is also
being studied in Southampton, however, fractionation procedures are not
envisaged.
2. C. T. M. C. Funded Stud' es with Cr. Ecobichon at McGl 1 I University
(i) Work will continue on the investigation of the effect of
smoke exposure on perinatal animals. These studies are aimed
at seeing whether exposure to cigarette smoke results in smoke
components reaching the pups, and if so. whether via the placenta
or breast milk. Arylhvdrocarbon hydroxylase will be used as the
biocneinical indicator.
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(ii) A programme of work on exposure of animals to ambient smoke
has been initiated. This programme will also use ary1hydrocarbon
hydroxylase as the indicator.
OM Work to develop a micronucleus test is anticipated. The object
is to obtain a short-term site specific test for tobacco smoke.
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BATCo document for Province of British Columbia 19 April 1999