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RESEARCH DEPARTMENT
Analytical Report
No. AR 89-04-132
KAI* K ~Kand Export) U ill ited
AESEA11 It & DEVELOPMENT CENME
YCAITil-irroj
Subject Ames mutagenic Activity of Smoke
Condensate from Cigarettes
with Modified Paper Characteristics
AuthorS Dr. E.D. Massey
TO Dr. T. Hirji
Circulation Dr- R.R. Baker
Mr. T.G. Mitchell
Mr. P.D. Case
File 46D
Date 3 May 1989
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BATCo document for Province of British Columbia 28
October 1999


Ames Mutagenic Activity of Smoke Condensate from
Cigarettes
with Modified Paper Characteri s tics
The purpose of the experiment was to investigate
the influence on the Ames
mutagenic activity of mainstream cigarette smoke
condensate (CSC) of
modifications to cigarette papers.
Cigarettes and Smokinq
Cigarettes ith a 59mm tobacco rod and 24.75mm
circumference using a U2TI/I
cellulose acetate filter and blend Iff-12 (a
modified flue-cured blend with
307. Btirley tobaccos) were manufactured using the
cigarette papers descrihed
in Table 1. Additionally, the R.J. Reynolds
product Vantage Excel was
taken and modified into the same construction as
the other cigarettes in
the series. This was achieved by removing the
tobacco, the rod length
being cut down to 59nNn and then refabricated with
the U1,12 blend. Finally.
the filter as removed and replaced with the
U2171/1 cellulose acetate
filter. As a control to determine if refabrication
modified the mutagenic
activity of CSC cioarette 062 was also
refabricated.
BATUKE R&D reference cigarettes A279 (flue-cured)
and A283 (US-style) were
also included in the experiment. CSC from A283 is
known to have higher
specific Ames activity than that of A279.
Sufficient cigarettes to produce approximately
150ing particulate matter
were smoked in a Borgwaldt RM20/CS smoking engine
to the specification of
the UK Laboratory of the Government Chemist. On
completion of the smoking
25% of the Cambridge filter pad was cut out with
the aid of a steel
template and used ior water and nicutine analysis.
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BATCO document for Province of BritiSh Columbia 28
October '1999


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Cigarette Code
Paper Code
Basis Wt.gm-2
Paper Permeability Coresta Units
% Calcium Carbonate
% Magnesium Oxide
Z Burn Additive
Table I Ciqarette Paper Specifications
D107 D61 D62 D63 D112 C501 C502 Vantage Excel
304P 5,4DEAO 5.4DEA2.5 5.4DEA5 5.4DEA5-EP
TOO-04403 TOD-04405
25.4 36.6 37.9 37.8 37.0 45.8 48.2 45.0
4 7 7 5 64 18 19 160
20.4 4.9 5.2 4.9 5.0 5.0 4.7 14.0
- 10.5 11.5 11.8 10.3 25.6 24.9 16.0
0. 5HAP - 2.0 flak 4.7 NaAc 2.1 MaAc 1.8 HaAc 4.6
NaAc 1.8 NaAc
3.0 KAc
MAP - monoammonium phosphate
NaAc - sodium acetate
KAc - potassium acetate
2 Z699600P


Ames Tests and Numerical Analysis
Followinc extraction of mainstream CSC from the
3/4 Cambridqe filter
Yads with dimethylsulphoxide. they were coded and
sent to Toxicol
Laborator4es. Ledbury, Herefordshire. for Ames
testing using
Salmonella typhimurium TA98 in the presence of a
10Z Aroclor 1254-
induced S9 system. The assay was carried out an
five separate
occasions using five dose levels of condensate and
triplicate
bacteria] platings per dose. The data were
evaluated using a
regressic.-i model to rank the condensates and to
provide an estimate
of the mutagenic ratio.
Results
The summary data are shown in Figures 1. 2 and
Table 2.
The data shows CSC from A283 too have a higher
activity than that
from A279 hich is consistent with historical data
(Figure 1. Table
2). Refabricating a cigarette did not seem to have
an effect on the
Ames mutacenic activity of CSC as the CSC from a
refabricated
cigarett-2 did not differ in activity from that of
a non refabricated
cigarette (compare D62 vs D62 refabricated (Table
2)). This
indicatec that the procedure adopted for the
assessment of Vantage
Excel was satisfactory.
Compariscri of the product produced with
conventional materials (D107)
with othcr cioarettes in the series. i.e. those
containing high
levels o' -macnesium oxide and/or the burn
additives sodium or
potassiu.-- acetate, indicates that there is no
significant effect of
these acents upon the specific Ames mutagenic
activity of the
resulting CSCs. (Ficure 2. Table 2).
The US-style reference cigarette, A283, whilst not
matched in design
characteristics to the experimental cigarettes
produced CSC of
similar activity to the experimental products
(Table 2).
The flue--t.-ired reference cioarette. A279,
produced a CSC of a lower
activitv than the experimental samples that would
be consistent with
the know- tobacco blend differences between these
products (Table 2).
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BATCo document for Province of BritiSh Columbia 28
October 1999


Table 2
Ames mutagenic ratios of mainstream CSC from
products with modified paper characteristics
Cigarette Ames* 95%
Mutacenic Confidence Limits
Ratio
D107 1.00
061 1.03 1.07
062 1.02 1.07
D62 refabricated 1.10 1.14
D63 1.12 1.17
D112 1.05 1.10
C501 1.14 1.20
C502 1.14 1.19
Vantage Excel 1.06 1.11
A279 0.79 0.82
A283 1.05 1.10
D107 independently assigned a value of unity
0.98
0.98
1.05
1.07
1.01
1.09
1.09
1.01
0.75
1.00
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BATCo document for Province of BritiSh Columbia 28
October 1999


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BATCo document for Province of British Columbia 28
October 1999