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T-7907 updating of the ITL Computerized-Cigarette-Design Program
Oblective:
To provide Product Development with a computer program to assist in the design
of cigarettes.
Status:
The ITL cigarette design program has been re-written for the IBM XT.
Preliminary testing with dummy data is complete and gives results identical to
the Apple version. We are now rewriting the brand data files and the software
required to update them.
Future plans for this system are to integrate the cigarette design program
with the data files generated by Analytical Services concerning current
cigarette deliveries and those generated by Product Development concerning
cigarette specifications.
T-7098 Design Guidelines for the Control of Puff Number
Obiective:
To produce a set of guidelines to be used in product design to control puff
number.
Status:
The first step of this project currently underway consists of reviewing past
ITL work and BAT group research in this area. The relative importance in the
control of puff number of parameters such as circumference, cuts per inch,
moisture content, paper porosity, paper additives, tobacco packing density,
etc. will be reviewed in the literature and assessed for further experimental
work. A tentative completion date has been set for September 1986. N-0
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T-7099 Evaluation of Reduced Sidestream and Self-Extinguishing Cigarettes
ObJective:
To assess, both objectively and subjectively, different methods of reducing
sidestream from cigarettes and to monitor developments in self-extinguishing
cigarettes.
Status:
As reported in the previous status review, low sidestream cigarettes made with
MgO paper and designed to deliver 4 mg of mainstream tar, were found to have a
similar taste character to control cigarettes made with standard paper.
However low sidestream cigarettes designed to give higher mainstream
deliveries (9 mg) had pronounced off-taste and irritation.
In an attempt to alleviate these problems, we have injected some of the
cigarettes (both 4 and 9 mg) with 'Black Tea Extract' at a level of 100 ppm.
Compared with the control cigarettes, the flavour injected samples were found
to be less irritating and had an Improved (but still essentially flue-cured)
flavour.
We are extending work in this area by making up 23 mm circumference cigarettes
from tobacco sprayed with 'Black Tea Extract' at a level of 100 ppm. The
cigarettes will have MgO papers - one from de Maud (DAC 10) and two from
Ecusta (18145 and TOO 02517, the latter being a high porosity paper containing
eucalyptus fibres).
We have recently received two new low sidestream papers from Kimberly-Clark.
We plan to test these papers for sidestream reduction as well as their effects
on taste and mainstream deliveries.
BATCo document for Province of British Columbia 19 April 1999

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T-6864 Alternatives for Saccharin in Cigar Tips
ObJectivet:
To define the most likely candidate as an alternative sweetener for cigar tips
and to evaluate It in terms of uniformity and method of application,
subjective characteristics and shelf life.
Status:
Aspartame has been identified as the best alternative to saccharin.
Large scale consumer subjective testing involved the preparation of 4 batches
of 4000 sweetened Colts Mild tips. Three levels of coating were chosen:
0. 03%, 0.04% and 0.05% (by weight) in order to determine which level most
closely matched the current on-market Colts Mild tips in terms of subjective
characteristics. Once an appropriate level is determined, the coating method
will be scaled up to industrial levels.
T-7087 Design of Cigarettes with Reduced Carbon Monoxide:Tar Ratios
Obiectlve:
To evaluate design options for reducing carbon monoxide:tar ratios from
cigarettes to less than unity.
Status:
Mattina of Kimberly-Clark has reported that alkali metal carboxylates added to
reconstituted sheet at levels approaching 10% have the effect of reducing
carbon monoxide deliveries when incorporated into cigarettes. Although
MaWna had no explanation for this, It is suspected that the reduction in
BATCo document for Province of British Columbia 19 April 1999

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carbon monoxide is similar to that previously observed when alkali metal
carbonates are added to stem, since alkali metal carboxylates decompose to
carbonates at relatively low temperatures. For cigarettes made with stem
containing more than about 3-4% sodium carbonate large increases in pressure
drop occur after lighting. This causes increased ventilation through the
paper and hence a decrease In carbon monoxide (and also deliveries of other
smoke components).
We have repeated Matti na I work using lamina instead of reconstituted tobacco,
with sodium citrate added at levels of 0, 5 and 10% (w/w). Smoke deliveries
were all reduced as sodium citrate levels increased. At 10% inclusion of
sodium citrate, carbon monoxide and tar deliveries were both reduced by about
20% and nicotine by 13%. Carbon monoxide:tar ratios were unchanged while puff
number increased by about 151/10.
Both the unlit and average lit pressure drops decreased with increasing sodium
citrate level. However the differences between unlit and lit pressure drops
Increased dramatically with citrate level: for instance the pressure drop
increase on lighting the cigarette with 10% sodium citrate was about 100%
greater than that obtained when the control cigarette was lit. At the same
time there was a significant reduction in both puffing and smouldering
temperatures.
These results indicate that sodium citrate causes a reduction in smoke
deliveries by increasing coal resistance and hence cigarette ventilation.
However since the carbon monoxide:tar ratios are not changed there appears to
be little practical application of this effect.
Reduction of carbon monoxide by diffusion through porous membranes is also
being investigated in an exploratory fashion. By a suitable choice of
membrane (Teflon or cellulose with pore sizes less than 0.01 um) diffusion can
occur without appreciable dilution of the smoke. Incorporation of such a
membrane into a filter, if feasible, will cause a considerable reduction in
the carbon monoxide:tar ratios.
BATCo document for Province of British Columbia 19 April 1999

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T-6140 Investigation of the Feasibility of On-Line Measurements of Tobacco
Filling Capacity/Density as a Process Development Tool
Objective,
To investigate whether a rapid and reliable method can be developed to monitor
tobacco filling capacity and/or density during processing.
Status:
It has already been shown (ITL Progress Report June 1985) that tobacco density
can be monitored at different stages of processing by the Tobacco Impedance
Gauge (T. I G.). At the present time a study of whether the T. I. G. and the
Borgwaldt densimeter can differentiate between various processing conditions
is being conducted. In a preliminary experiment, the drying conditions were
systematically varied using 'process air* as a means of control.
A V-23 + PCL blend was processed under low, medium and high drying conditions
(296, 1471 and 5500 grain loadings). Samples were taken before and after the
cutter, after the dryer and after the cooler. Measurements of impedance.
filling capacity and moisture content were carried out on unconditioned
samples in the Pilot Plant and on conditioned samples in the laboratory.
Results Indicate the following:
1) Impedance increases with the degree of drying,
2) the dryer has the greatest effect on impedance,
3) the impedance of the Pilot Plant samples are smaller than the laboratory
samples.
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Filling capacity results appeared to be very moisture dependent. With the
Pilot Plant samples, a large Increase was observed after the dryer. After the
cooler the values were slightly less. The changes in filling capacity
obtained with the conditioned samples were much less than the changes observed
with the unconditioned samples, but both sets of samples follow the same
trends.
The T. 1. G. and Bor9waldt densimeter can, to a certain extent, differentiate
between various drying conditions. To better assess these two instruments as
process development tools, the tobacco impedance and filling capacity will be
measured as a function of time of processing. The effect of three different
processing conditions (1000, 1500 and 2000 grain leading of the exhaust
process air) will also be investigated. Pressure drop and firmness will be
measured for cigarettes made from the same tobacco in order to relate these
cigarette characteristics to the cut tobacco properties.
Consideration will be given to investigating the effect of processing on
tobacco structure.
T-6127 Investigation of the Relationship Between Cigarette Firmness and
Tobacco Properties
Objective:
To develop a model to predict cigarette firmness from tobacco properties and
to identify any tobacco properties which can be commercially modified to
improve cigarette firmness.
Status:
The Initial step of this project is to review work carried out in that area of
research. Tobacco particle density, tobacco particle properties, tobacco
processing and tobacco density are the factors currently being reviewed. The
review is expected to be completed by July 1986.
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T-0190: Non-routine Analytical Service to Support R & D Pro-iects and for
General Troubleshooting
Objective:
To meet the analytical requests made by different services such as:
Purchasing/Technical Services, R & D, Marketing, Manufacturing/Engineering.
Status:
Many of the requests require non-routine analytical methods, which often
Involve literature searches, method development and multiple instrumental
techniques.
During the past six months, the main areas of activity have been:
1) Quality Control of Menthol Crystals for Guelph Plant-
Random samples are taken from every new shipment of menthol crystals and
are analysed by G. C. to detect any trace of Impurity.
2) Follow-up of Humectant Levels in Opposition Brands:
Glycerol and Propylene Glycol are regularly measured on five RJR-MacDonald
brands to maintain an awareness of opposition trends.
3) Analysis of Cigar Tips for Saccharin:
110@ The saccharin level of new batches of sweetened tips is measured to ensure
that it remains within specifications (approximately 5% by weight of total
saccharin).
4) Determination of LiCt in PCL:
LiCt has been added to PCL during manufacture of experimental samples.
This is used as a tracer to assess PCL concentration in finished blends.
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5) Menthol Levels in Tobacco and Vgarettes:
Recent product development projects have required tobacco to be sprayed
with menthol prior to cigarette manufacture. Menthol levels after
spraying and during manufacture were determined in order to assess any
loss of menthol (22%-25% by weight).
6) Consumer Complaint Investigation:
A taint problem on Cameo Ultra Mild cigarettes was identified as being due
to a spill of SUNAPLEX 992E, a grease used in cleaning cigarette packing
machines.
7) Individual Alkaloid Determination in Tobacco Samples:
A G. C. method has been set up to measure the nornicotine content of
certain tobaccos. Levels exceeding 15% of the total alkaloid content have
been associated with poor smoking quality.
T-9050 Determination of Environmental Contaminants In Plant Working Areas and
External Atmosphere
Oblective:
To check the concentrations of menthol and nicotine in plant atmospheres on a
regular basis to ensure that the levels meet safety guidelines.
Status:
Approximately once a month during the processing season samples are received
from Aylmer Plant to be analysed for their nicotine content. These samples
consist of small cartridges filled with an appropriate sorbent which retains
nicotine when ambient air is pumped through. During 1985, the nicotine levels
did not exceed the TWA (0.5 mg /M3 and STEL (1.5 mg/m3 limits, except
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when Burley tobacco was processed. In the latter Instance levels as high as
3.4 mg/m3 were measured. The external environment is monitored at Ajax
Plant:
1) by collecting air samples inside the chimney.
2) by measuring the nicotine level of waste water used for the treatment of
tobacco.
The levels were in the range: 0.2 mg/m 3 to 3.9 mg/m3 for air samples and
0.6 mg/m,3 for water samples. There are no special regulations concerning
the allowable nicotine levels of these external samples.
T-6967 Sidestream Smoke and Irritation
Oblective:
To determine which components of sIdestream smoke are the main contributors to
irritation with a view to reducing their concentration.
Status:
A special collection system for sidestream smoke was designed. It is a two
part, all glass. pear-shaped apparatus. Two openings, drilled into opposing
walls of the lower part of the apparatus allow the cigarette to be placed into
the center of the vessel and to be lit. The upper portion is connected at one
end to a vacuum pump which draws air through the system and at the other end,
via a 3 cm diameter connecting hole. to the lower portion of the apparatus. A
Cambridge filter (d - 3 cm)-is placed between the upper and lower portions
(two parts) which are then sealed with small clips. The ai rf low through the
chamber is adjusted to give I ami nar flow thus minimizing condensation onto the
chamber walls.
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Analytical methods have been set up to quantify potential irritants such as
low molecular weight carbanyls and bases. The procedures are as follows.
Particulate matter in both M/S and S/S Is trapped on a conventional Cambridge
filter assembly while the vapour phase is collected in traps placed after the
filter assemblies.
The carbonyls are derivatized directly in the trap with 2-4 Dinitrophenyl-
hydrazine. After extraction and clean up, the 2-4 Dinitraphenylhydrazine
derivatives are separated and quantified using Reverse Phase High Performance
Liquid Chromatography and UV detection at 360 nm. This method provides
sensitive and reliable results and will be a very useful tool for the
screening of different cigarettes in terms of their carbonyl content.
The medium and high boiling point bases are trapped in an acidic methanol
solution. concentrated, released via neutralization and directly injected onto
a capillary gas chromatograph equipped with a Nitrogen specific detector. Due
to the limited number of pure standards available, only 32 bases have been
identified and quantified. The results are quite reproducible and in good
agreement with literature values.
Literature an In vitro Irritation tests has also been reviewed and a few
methods using cell cultures show promise.
The next step in this study is to determine carbonyls and bases in sidestream
smoke from different brands of cigarettes, and to see whether their
concentration can be correlated in any way with perceived irritation.
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T-7084 Fractionation of-Cigarette Smoke by Liquid Chromatographic Methods
Obiective:
To develop an efficient and rapid method to fractionate cigarette smoke for
the determination of the Ames mutagenicity of the individual fractions. The
aim of this work Is to compare the mutagenicity of the individual fractions
relative to one another and to whole smoke. The long term purpose of this
work Is to investigate the feasibility of developing products with reduced
biological activity. This study Is being carried out in conjunction with the
Biological Research Group.
Status:
Conventional liquid-liquid fractionation methods have been used in the past to
divide cigarette smoke concentrate into different fractions. However, these
methods were long, tedious and not adaptable to any large scale study.
A novel approach to complex mixture fractionation is to use HPLC columns. The
first stage of this project Is currently under way and involves adaptation of
the HPLC method to the special composition of cigarette smoke concentrate.
The following chromatographic parameters are being optimized:
1) Solvent composition.
2) Flow rates.
3) Recovery of fractions.
Once this preliminary part has been completed (June 1986) the method will be
scaled-up for the semi-preparative work. Smoke fractions will then be
prepared on a daily basis and given to the Biological Group for mutagenicity
assessment.
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T_S549 Development of High Pressure Liguid Chromatographic Methods
Obiective:
To develop specific analytical methods for non-volatile or heat sensitive
compounds and provide cost-and time saving alternatives for presently used
methods.
Status:
To date, methods have been developed for analyses of:
- Aspartame on cigar tips.
- Saccharin in cigar tips.
- Glycerol in tobacco.
- Low molecular weight carbonyls in cigarette smoke.
For every new method, the following steps have been followed:
1) Review the literature.
2) Optimization of the different analytical parameters.
3) Overall evaluation.
4) Writing the procedure for routine use.
This is an on-going project, and other methods will be implemented as new
needs are expressed.
T-6966 AD21ication of Methods for Measuring Sidestream Smoke Deliveries
9tilctive-
To develop methods for the collection and analysis of s1destream smoke.
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Status:
We have recently acquired a new MOIR CO/CO 2 meter (Analygas) for determining
these gases in sidestream smoke. Testing of this new meter has shown that the
older Filtrona CO meter does not have a similar response curve. The new meter
gives lower readings at low CO concentrations (as in sidestream) and higher CO
readings at high concentrations (as in mainstream) than the Fl I trona. In. fact
sidestream carbon monoxide concentrations given by the new meter are about 20%
lower than those given by the Filtrona.
We are currently designing an integrator for the CO/CO 2 meter so that total
masses of CO and CO emitted in the sidestream, can be determined over the
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total burn time of the cigarette. This system will also be used in the paper
combustion experiments (T-7279-1).
T-9051 Updating the AnalXtical Methods According to New Safety Guidelines
Oblective:
To supplement all currently used analytical methods with the appropriate
safety guidelines.
Status:
A short, clear safety guideline is being added at the beginning of each method
to help increase worker awareness of the proper safety precautions. This is
to comply with new laws on safety in chemical laboratories. Out of a total
number of 24, nine methods, five in English and four in French. have already
been reviewed.
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T-8079-1 Program for Upgrading the Recording/Dup1tcating System for Human
Smoking
Obiective:
To ensure that all aspects of human smoking are accurately and properly
monitored.
Status:
This is an ongoing activity to upgrade the recording and duplicating system
for monitoring human smoking behaviour.
The strategy is to replace the current ISC microprocessor used for duplication
with an Apple Ile/Isaac system. This will result in a fully Integrated
recording/duplicating system.
To date, phase 1 of the initial proposal has been achieved. This phase
consisted of writing and testing control procedures for the Isaac's data
acqu'sAtlcn and control of the puffing process.
Phase 2 is underway and will be finished by February 15, 1986. This phase
consists of using simulated puffs to test the software for control of the
stepping motors and switches. Tests on a prototype (with I piston) are also
underway.
The final phase is the validation procedure. This includes tests with real
data which should be completed by the beginning of March 1986. The new smoke
duplicator will then be extensively evaluated in terms of accuracy and
reproducibility.
Consideration is also being given to the feasibility of interfacing the Apple
He with the mainframe computer. This will allow us to transfer data directly
from the Apple to the mainframe so that SAS statistical packages may be more
readily accessed.
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BATCo document for Province of British Columbia 19 April 1999

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T-8079-2 Alternative methods for Duplication
Mective:
To review the presently available methods, both within and external to the BAT
group, for measuring mouth level smoke constituents with the aim of assessing
the feasibility of adapting and/Dr developing a method to be used
alternatively or In conjunction with duplication.
Status:
Two methods were designated for assessment:
1) the BAT (UK & E) R & 0 computer program DELIVER.
2) the determination of puff by puff TPM by IR reflectance.
A preliminary investigation of the applicability of DELIVER to ITL has been
completed. The results of this investigation indicated that DELIVER was able
to predict total deliveries of tar and nicotine as determined by standard
smoking conditions to within 1.00% and 1.25% of the experimental values
respectively. However, this could only be achieved if some of the input
parameters were significantly altered from the original values. Furthermore
there appeared to be serious discrepancies between predicted (DELIVER) and
experimental values on a puff by puff basis. Differences ranged from 13-100%
higher for the first puffs and 4-16% lower for the last puffs. This
investigation is presented in an internal report (November 30, 1985, S.
Ke I eman) . The results have also been communicated to BAT (UK & E) R & D so
that further refinement of the model may continue. Based on current
Information it is not recommended that DELIVER, in its present form, be
incorporated into the Human Smoking Behaviour methodology. However should the
model be sufficiently refined in the future, serious consideration should be
given to its inclusion as a Human Smoking Behaviour procedure.
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BATCo document for Province of British Columbia 19 April 1999

The feasibility of incorporating the determination of puff by puff TPM by IR
reflectance into the recording equipment for Human Smoking is presently being
assessed. Based on the method of Jenkins and Gale I an initial apparatus was
developed by BAT UK & E R & 0 during 1983-84. Before any significant
investigation is initiated, the work of BAT UK & E R & D in this area will be
reviewed. If these results seem promising, the apparatus could be developed
during 1986.
T-8079-3 Assessment of the Feasibility of measuring Inhalation Patterns and
Human Retention of Cigarette Smoke
Oblective:
To Investigate the usefulness and feasibility of measuring human inhalation
patterns and retention of smoke for possible inclusion into human smoking
behaviour studies.
Status:
The project was initiated In Octoter 1985. To date a report concerning this
subject has been completed.
This report is a review of methods developed both within and external to the
BAT group for the measurement of human inhalation and retention of cigarette
smoke.
1 Behaviour Research Methods, Instr. & Computers, 1984, 16(3),
263-267. 'An Instrumental Cigarette Smoke Monitor Designed for the
Direct Measurement of Smoke Particulate Matter Generated in Human
Smoking Studies".
BATCo document for Province of British Columbia 19 April 1999

19
It is made up of three parts:
a) Smoking patterns (Inhalation/Exhalation) and puff parameters.
b) Human retention of cigarette smoke (nicotine and tar).
C) Human retention of carbon monoxide from cigarette smoking.
The conclusions and recommendations are summarized as follows:
Of all the methods reviewed in this project. only the chest respiratory
inductive plethysmograph combined with a modified cigarette holder or a cheek
plethysmograph appears to be feasible for use in Human Smoking studies. This
method would allow the measurement of both Inhalation and puffing parameters.
The results obtained appear to be relatively accurate, reliable and
non-obtrusive. Although quite expensive, this method permits measurements in
a "natural" environment. However, the decision to Include such a method in
Human Smoking Research will also be based on the relative value of the new
information which would be added by the method.
Many methods have been developed for measuring smoke retention. Experimental
procedures are similar throughout all research groups and implementation
appears to be relatively simple. The same provisos mentioned above for
inhalation also apply to the adoption of a method for retention.
T-8077-17 Investigation of the Effects of Nicotine Enhancement on Human
Smoking Behaviour
Objective,
To assess the changes, if any, occurring in Human Smoking Behaviour as a
result of cigarettes with enhanced levels of nicotine.
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Status:
As a result of the pilot study conducted in April 1985, the appropriate levels
of nicotine enhancement were determined to be 15 and 30Y* for mid to high
delivery products and 30 and 45% for low delivery products. These levels are
sufficiently high that changes in smoking behaviour and human delivery can be
attributed to increases In available nicotine and not to variability in-the
apparatus.
Du Maurier K. S. and Matinde E.M. K. S. (15 and 4 mg tar delivery respectively)
were selected as the brands for study. Cigarettes were injected with the
appropriate quantities of nicotine citrate solution to achieve increases in
smoke nicotine content of 15 and 30% for du Maurier K. S. and 30 and 45% for
Matinde E. M. K.S. The du Maurier K. S. samples had to be rejected since a
distinct off-taste was noticed during subjective evaluation. Panelists had
difficulty in differentiating between the control Mat -E.. M. K. S. and the
nicotine enhanced samples. The leaf and smoke analyses were conducted
concurrently with the subjective evaluation. Leaf analysis indicated the
appropriate increases in blend nicotine. However, smoke analysis showed a
poor transfer of injected nicotine from tobacco to smoke. This may have been
due to an uneven distribution of nicotine along the tobacco rod when the
samples were injected with nicotine citrate. It may have also contributed to
the off-taste found for du Maurier K. S.
In light of the above. small quantities of Matin&e E. M. K.S., du Maurler K.S.
and du Maurier Light K. S. tobaccos were sprayed with sufficient nicotine
citrate to increase the smoke level of nicotine by 15, 30 and 45%. Smoke
analysis results indicated that the smoke nicotine of these experimental
cigarettes was appropriately increased.
BATCo document for Province of British Columbia 19 April 1999

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Subjective evaluation of the Matinte E.M. samples indicated a small but
definite difference between the control and enhanced nicotine cigarettes. The
nicotine enhanced cigarettes were perceived to have more impact and to be
slightly more Irritating than the controls, but no off-taste was detected. du
Maurier K.S. cigarettes with 30% more nicotine were also tested subjectively.
These.results showed du Maurier K.S. with added nicotine to have an equal or
slightly greater taste level, more impact and to be slightly more irritating
than the control. Subjective evaluation of the du Maurier Light K.S. clearly
showed an increase in taste, Impact and irritation of the product with an
increase in nicotine level. Of the three brands tested du Maurier Light K.S.
appeared to be the most acceptable and will therefore be the product used for
further study. The human smoking study is expected to start in April 1986.
T-8082-1 Application of Delta for Product Screening and Evaluation
a) Project Omega
ObJective:
To determine the relative positions of Matinie K.S. and Regular in the market.
Status:
Matinee, in both king size and regular lengths, was evaluated by the Delta
panel relative to the major competitors. For both lengths, subjective
evaluation indicated that Matinde was milder than its competitors, having less
irritation, taste and after-effects. For the king size brands. Craven A
(Rothmans) appeared to be most similar to Matinie, while for the regular
length brands, Belvedere E.M. (8 & H) was the closest match to Matin6e.
BATCO document for Province of British Columbia 19 April 1999

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b) Project Swirl
Obiective:
To further evaluate the subjective characteristics of two novel filters
designed to improve the smoke quality of low delivery cigarettes by changing
the smoke pattern.
Status:
Due to differences in filtration efficiencies and ventilation, direct
comparison of swirl filters and regular cellulose acetate filters on the
subjective smoke qualities of Medallion was not possible. For this reason the
contribution of swirl to subjective improvement was assessed by comparing
swirl filters both with and without a plastic sleeve (which disrupts swirl).
In the previous progress report (May 1985) it was noted that the sleeve itself
caused a subjective change. The sleeve has now been redesigned and using the
new sleeve it has been shown that swirl does improve the subjective response
to Medallion, which delivers nominally 1 mg of tar. The next phase will
consist of measuring the deliveries of these products under both standard and
human smoking conditions.
T-8082-3 Investigation of the Effect of Brand Imagery on -SubJective Evaluation
gkilcti ye:
To study the influence of brand identification and pack imagery on subjective
evaluation.
BATCo document for Province of British Columbia 19 April 1999

23
Status:
The Initial study was reported in the previous progress report and details are
given in an internal report (June 1985, C. Boisjoly) . A second study was
designed to:
1) control for product familiarity by using foreign products.
2) isolate the pack effect from the brand name effect by evaluating empty
packages and subsequently masked products In different packages.
Panelists were asked to evaluate:
1) empty packages in terms of product expectations.
2) masked products alone.
3) masked products presented In their own packages.
4) masked products in different packages.
5) unmasked products alone.
At this point the first four stages have been completed. Each evaluation was
repeated twice with an inter-evaluation interval of two weeks. In any of the
evaluations 85-90% of the variation has been explained and the inter-trial
correlation has exceeded 0. 80.
The fifth and final phase is presently underway and will be completed by the
end of May.
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T-8082-4 Investigation of Product Quality Attributes
Objective:
To determine the threshold level at which subjects start to perceive a
difference in the firmness of cigarettes.
Status:
Since the initial study investigating cigarettes differing in weight by + 50
mg, a new study was initiated investigating cigarettes with weight differences
of + 25 mg. Results indicate that, an average, subjects were still able to
differentiate the firmness between the cigarettes, despite the fact that
approximately 25% of the respondents claimed their answers were guesses and
all found the test very difficult. It appears, therefore, although the group
results indicate that subjects can perceive tactile differences, a smoker
would probably not spontaneously recognize this difference as he picks a
cigarette out of pack. Therefore It is suggested to further study the impact
of these firmness changes on the sensory evaluation of the product.
BATCo document for Province of British Columbia 19 April 1999

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T-7708: Mutagenicity Studies
ObJective:
To determine those parameters which affect mutagenicity of smoke condensate,
and, by monitoring market brands for condensate mutagenicity on a periodic
basis. to help ensure that ITL brands rank favourably.
Status:
1. Mutagenicity of Commercial Cigarettes
Since the last status report, we have extended our studies to include more
N. American and European brands. In an examination of the mutagenicity of
a group of low delivery, ventilated Canadian brands, it was found that a
particular opposition brand was significantly lower than 3 other brands,
produced by ITL and Rothmans. This result is interesting since our
earlier studies showed that a brand from the same competitor had the
lowest mutagenicity among a group of 26 N. American brands.
In a recent examination of a French brand we speculated that the version
of this brand available in Canada appeared to be lower in mutagenicity
than similar cigarettes available here a few years ago due to the former
being manufactured with significantly lower amounts of dark fermented
tobacco. Consequently, we tested two French brands available In Canada
with those available in France, and found that for one brand there was no
difference in activity, while for the other brand there was a significant
difference. This suggests that the Canadian version of the latter brand
contains significant amounts of a lower activity tobacco (possibly
flue-cured).
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All our studies have shown that French cigarettes containing dark
fermented tobaccos are much higher in activity than blended U.S. brands.
Other European countries such as Spain and Portugal appear to use similar
tobacco in their cigarettes and therefore, we obtained and examined one
brand from each of these countries. This study showed that the Spanish
cigarette was much higher in activity than the Portuguese cigarette which
was similar in activity to a U.S. blended cigarette. Both brands had
significantly higher activities than Canadian brands containing only
flue-cured tobacco.
2. Mutagenicity of Condensate from Player's Cigarettes Modified by Electro
Medica Office
Mutagenicity testing of these cigarettes along with market samples of the
same brand showed that the two were similar in activity, and that the
Electro Medica modification does not affect the Ames biological activity.
3. A Comparison of Rat and Guinea Pig Metabolic Activation Systems
The standard Ames test employs Aroclor-treated rat liver post-
mitochondrial supernatant (S-9) as the activation system. Earlier studies
in this laboratory suggested that a similar preparation from the guinea
pig was more active in converting cigarette smoke condensate to mutagenic
intermediates In the Ames test. Preliminary results obtained suggest that
the initial observation is true and that the concentration of S-9 might
play a role in the level of activation obtained.
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4 . Selenium and Cigarette Smoke Condensate (CSC) Mutagenicity
Selenium as sodium selenite has been shown to be an effective inhibitor of
the mutagenic activity of several chemicals such as 2-acetylaminofluorene,
2-hydroxyacetylamino flucrene, and 7,12-dimethylbenzanthracene, which
require metabolic activation before they manifest their activity. It Is
also known to reduce the mutagenic activity of chemicals such as
N-methyl-NI-nitro-N-nltrosoguanidine, malonaldehyde and propiolactone
which are direct acting mutagens. Hence, we thought it useful to
determine If selenium could reduce the mutagenicity of CSC.
Preliminary studies undertaken using Ames strain TA-98 suggest that sodium
selenite can reduce the mutagenicity of CSC, while sodium selenate can
increase the activity of CSC. The increase in activity appears to be due
to the positive mutagenicity of sodium selenate itself with strains that
detect frameshift mutagens. The reduction in mutagenicity of CSC by
selenite is an interesting observation and more work is planned to confirm
it.
T-677S Microbiological Examination of Tobacco Products
Objective:
Fall To survey the microflora in our own and opposition tobacco products.
Status:
1. Ci-garettes
Most of the 7 brands selected were examined by B.A.T. Southampton in
1982-83. In that study the total bacterial counts for the Canadian
cigarettes were found to be an order of magnitude higher than the counts
obtained for cigarettes of U.K. manufacture. The present Montreal study
of Canadian cigarettes found similar levels of bacteria to those found by
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Southampton two years earlier, indicating a consistency in the handling of
the tobacco during cigarette manufacture.
Of interest is the fact that the two brands with the highest bacterial
counts also showed the lowest specific mutagenic activity in the Ames
test. Hence one may speculate that similar grades of tobaccos are used
for these brands. Whether the microorganisms themselves have any role in
lowering mutagenicity Is unknown.
The main difference between this ITL study and the Southampton examination
of 2 years ago was the lower numbers of thermophilic actinomycetes
recorded in the former study. This would suggest that more care has been
taken in the handling of tobacco or-in cigarette manufacturing in the last
2 years. There was nothing remarkable about the co I if orm bacteria, which
were not detected, nor in the mould content of the cigarettes.
2. Fine Cut Tobaccos
Eight samples were selected for this study which included I of the 3
brands examined by Southampton in 1982-83. The bacterial levels recorded
in this ITL study of 24 samples from 8 brands were about three times
higher than those recorded In 1982-83. However, a direct comparison
cannot be made on account of the different samples examined as well as the
small number tested earlier. As in the case of the cigarettes, the
Montreal study showed fewer actinomycetes In the fine cuts than the
Southampton study of 2 years ago, indicating a possible general
improvement in the handling and processing of the tobacco. Both the
Southampton and Montreal studies failed to detect co] if orm bacteria or
fungi in these products.
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3. Pipe Tobacco
An examination of 4 brands was undertaken by ITL. Old Port showed the
highest number of bacteria (9 x 10 6/gram) followed by MacBaren (3 x
106). The other two brands, Skandinavik and Sorkum Rif showed numbers
of bacteria 2-3 orders of magnitude lower.
A considerable number of thermophilic actinomycetes were also found in Old
Port (6.5 x 102). However. this number Is considerably lower than the
value of 1.9 x 104 recorded for this product 3 years earlier.
Skandinavik and Borkum Riff did not show the presence of thermophilic
actinomycetes. No collforms or fungi were detected in these products.
4. Cigars
In the 19B5 ITL study 12 brands of cigarillos and cigars were examined for
their microbial content. Ten of these had been examined by Southampton In
1982-B3 hence a good comparison can be made. O'Sherry Filter Tip (RJR)
examined earlier was not available in 1985. and it is not known whether El
Producto (RJR) examined by Southampton were cigars or cigarillos.
Bacterial counts for El Producto cigars and cigarillos were very similar
in both the ITL and Southampton studies. Once again Southampton recorded
much higher counts of thermophillc actinomycetes than did ITL.
While this is most probably due to a general Improvement in the handling
eo-'- and processing of tobacco, it Is also possible that differences in
microbiological procedures such as sampling, as well as accuracy of the
incubation temperature (509c) 0 account for the difference.
Neither study detected co I I form bacteria In any sample, and there was
nothing remarkable about the numbers of fungi in the ITL study.
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T-7710 Biochemical Effects of Tobacco Smoke Exposure in Experimental Animals
Objective:
To study the effect on protective mechanisms in experimental animals of
exposure to mainstream and sidestream cigarette smoke.
Status:
Cigarette smoke Is known to contain oxidants which could affect levels of
endogenous protective mechanisms in the lung. Studies have shown that when
animals are exposed to cigarette smoke there Is an Increase In levels of
glutathlone (GSH), an antioxidant present in significant quantities in
tissues. It has also been shown that when experimental animals are treated
with GSH -depleting chemicals such as diethylmaleate, the rapid depletion of
GSH is followed by a re-synthesis of this antioxidant. Such a re-synthesis of
GSH was not considered in the above mentioned studies involving cigarette
smoke. Hence it was considered important to determine whether exposure to
cigarette smoke would lead to an Immediate loss of GSH followed by a recovery
of normal levels such as occurring after treatment with diethylmaleate.
Adult male, Sprague-Dawley rats were exposed to dilute cigarette smoke using
the B.A.T.-Mason Inhalation System. Animals exposed to 40, 120 and 240 puffs
were sacrificed immediately after smoke exposure. The rats were euthanized
and tissues (lung, liver and kidney) were removed and used for determination
of their glutathione (GSH) content employing El lman I reagent (515-dithiobis
(2-nitrobenzoic acid)).
Exposure to 40 puffs of a 1:5 diluted smoke (3.33 minutes) resulted in a
slight lowering of GSH in the lung, but did not affect levels of this
antioxidant in the liver and kidney. Increasing smoke exposure to 120 puffs
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(10 minutes) resulted In a lowering of GSH in lung and kidney but not In the
liver. However, increasing the smoke exposure to 240 puffs (20 minutes)
caused a significant reduction of GSH In all three tissues. A reduction of
GSH in tissues of smoke-exposed rats is important because of the general
protective role of this antioxidant, and warrants extending our studies to
other antioxidants (Vitamin Q and other tissues (bladder and blood), as well
as other to species such as the guinea pig and hamster.
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BATCo document for Province of British Columbia 19 April 1999