ki. t;.-Mltchell/KO)
PROSPECTS FOR AUGMENTING NICOTINE CONTENT OF
TOBACCO PRODUCTS
1. Sources
a) Synthesis
b) Cultivation of Nicotiana
c) Plant Cell Culture
d) Waste tobacco or by-product from processes
e) Quantities
2. Incorporation and Augmentation
a) Use of high nicotine tobacco
b) Use of tobacco extracts
c) Use of pure nicotine
d) Augmentation of smoke nicotine
3. Activity by Competitors
4. Restriction on Use of Nicotine
5. Toxicological Aspects
6. Conclusions
1. SOURCES
There are three major alternative routes to obtain
nico-
tine, of which only one- the growth of plants of
species and
varieties of Nicotiana - has been exploited
commercially to a
significant extent. The routes are:
a) Synthesis
Methods of producing nicotine identical with the
form
present naturally in tobacco have been devised and
confirmed
as feasible in the past. The major barrier to the
use of this
approach in the past has always been of cost, with
the synthe-
tic material unable to compete with nicotine
obtained from C=)
extraction of tobacco or N.rustica. ON
co
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Novernber 1999


-2-
b) Cultivation of Wicotiana
Although a variety of plants have been shown to
produce
nicotine, only species of Nicotiana. have been
found to contain
it in large amounts. N.tabacum and N.rustica are
the main
sources and yields up to 8Z by dry weight of
leaves have been
recorded.
From published data, varieties can be regarded as
commer-
cially available which will produce on average:
Flue-cured tobacco
Burley tobacco
N.rustica (air cured)
4% nicotine or 100 kg/ha for a
yield of 2500 kg leaves/ha
5% nicotine or 125 kg/ha for a
yield of 2500 kg leaves/ha
8% nicotine or 160 kg/ha for a
yield of 2000 kg leaves/ha
To obtain nicotine by deliberate production of
'tobacco'
it would appear that N.rustica offers clear
advantages, but
two points require further consideration. The
first is that
a trial of N.rustica lines obtained world-wide and
conducted
in Canada showed a range of alkaloid content from
1.3 - 8.2%
so that it must be considered as inherently as
variable as
tobacco. The best variety reported in the Canadian
work was
of Italian origin (Brasile Selvaggio) with an
estimated yield
of 149 kg nicotine per hectare from 1816 kg of
leaves. The
second point is*that whilst highest nicotine yield
would be
the major consideration if the nicotine were to be
extracted,
blend compatability might restrict the choice to a
source with
a nominally lower level if the leaf were to be
incorporated as
cut tobacco or in sheet.
An additional variation on this theme would be the
selec-
tion of upper plant positions of bigh-nicotine
tobacco varieties
with levels of nicotine significantly above the 4
and 5% quoted
as an across-the-plant figure. The lower plant
positions would
still be utilisable in other blends, but this
argument could
not be applied to N.rustica.
N.)
C)
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November 1999


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c) Plant Cell Culture
The growth of isolated tobacco plant calls has
b*en
the subject of extensive studies by the Japanese
Monopoly,
ourselves and, to an uncertain extent, Gallahers.
Some of
this work has been concerned with producing
nicotine in which
it offers some theoretical and potential
advantages in relation
to purity, source as 'nature-identical', and a
safeguarding
of supply. Whilst apparently significant yields
have been
obtained on the basis of dry weight of tisiue,
enormous tech-
nological barriers would have to be overcome to
make it a
practical route for large-scale production of
nicotine. For
the foreseeable future, this area would have to be
regarded
mainly as a route to obtaining novel, but
expensive, tobacco-
based smoking materials or tobacco substances of
much higher
cost than nicotine. Exploitation would ultimately
require con-
siderable capital investment in a novel and highly
speculative
technology.
d) Waste tobacco or by-product from processes
Waste tobacco represents a potentially useful
source of
nicotine. Although the content of the latter is
usually rela-
tively low, it could be worth comparing the use of
waste i
for this purpose in comparison with their
reconstitution or
the cost of purchase of leaf solely for extraction
of the
nicotine..
Several extracts obtained in processing may be
expected
to contain nicotine, including those from del&rate
attempts
to-de-nicotinise such as the Taynag process, and
also from other
extraction processes (Hag, DIET). The value of
these would
depend on the quantities available and the
concentration of
nicotine in them.
e) Quantities of nicotine required
11 hatever the form or source of nicotine to be
considered,
it must be available in sufficient amounts to
sustain the use
of the fortified blend in question. Two simple
examples pro-
vide a guide to the relative-amounts involved:-
r".;
cc
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November 1999


AV
-4-
i) 1000 million cigarettes per annum, nominal
weight
1 g per cigarette, blend fortification from 2 to
2.52
nicotine.
Additional nicotine required: 5000 kg per annum.
From use of high nicotine tobacco (section 1b),
this
would require:
50 ha flue-cured tobacco producing 100 kg/ha
nicotine, or
40 ba Burley tobacco producing 125 kg/ha nicotine,
or
31 ba N.rustica tobacco producing 160 kg1ha
nicotine'.
ii) 1000 million cigarettes per annum, nominal
weight
1 g per cigarette containing 50:50 tobscco:tobacco
sub-
stitute. Blend nicotine 1% to be fortified to 2Z
would
require nicotine from 100 ha flue-cured, 80 ha
Burley or
62.5 ba N.rustica.
2. INCORPORATION & AUGMENTATION
The simplest route to nicotine fortification of
blends
would be the use of high nicotine leaf as a
substituent for
low nicotine portions of the blend, either as
whole leaf or
the use of extracts rich in nicotine for addition
to the whole
or part of the blend. These approaches, although
limited in
the extent to which the nicotine content overall
can be in-
creased, have considerable advantages over the use
of pure
nicotine in being more or less free of legal
restraint or
hazard to work force. A disadvantage is that high
nicotine
grades, as associated with upper plant positions,
are assoc-
iated with higher TPM deliveries and might be
expected to
affect other aspects of smoke properties and
perception
beyond that of nicotine alone.
a) Use of high nicotine tobacco
Two routes to the incorporation of high nicotine
leaf
are as cut tobacco or as a component of
reconstituted
tobacco. The simplest effect of substitution in a
blend of
average nicotine content 2%, assuming the material
which it
C:r\
was used in place of had the blend average
nicotine content,
would be:
co
BATCo document for Province of BritiSh Columbia 4
November 1999


-5-
Nicortine content (%) Nicotine conterit of Final
Blend for Substitution of
of added tobacco 5% 10% 15% 20%
4 2.1 2.2 2.3 2.4
5 2.15 2.3 2.45 2.6
6 2.2 2.4 2.6 2.8
7 2.25 2.5 2.75 3.0
8 2.3 2.6 2.9 3.2
9 2.35 2.7 3.05 3.4
10 2.4 2.8 3.2 3.6
In practice, the substitution of the lowest
nicotine
grades in the blend would slightly improve upon
these figures.
It is also clear, that to avoid blend distortion,
tobaccos
with very high nicotine contents would be required
to achieve
a useful degree of nicotine augmentation (e.g. 10%
- 25%).
The incorporation of high nicotine leaf in
reconstituted
tobacco would offer an advantage if it were used
as a replacement
in that the normal sheet material would be much
lower than
the blend average in nicotine content, e.g.:
'Normal Blend' 'Modified Blend'
CRS 15% @ 0.7% nicotine 'CRS 15% @ 0.7% nicotine
Sheet 5% @ 0.7% nicotine Sheet 5% N.rustica @ 6%
nicotine
Cut Tobacco 80% @ 2.3% nicotine Cut Tobacco 80% @
2.3% nicotine
Bl*end average: 2% Blend average 2.26%
In a 'high-sheet' blend, higher levels of
incorporation
would have increasing beneficial effects on the
final nicotine
content. In the above example, W.rustica sheet is
shown as
having a nicotine content of only 6% as it is
assumed a quarter
of the original would be lost in the sheet
process. In this
C)
respect, the Austrian reconstitution process would
be advan- CD
tageous in retaining practically all solubles, and
hence
nicotine.
(ZD
BATCo document for Province of BritiSh Columbia 4
November 1999


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An additional route toutilise high-nicotine leaf
would be in the form of a dust-coating to cut
tobacco as
examined some years ago at Southampton for
flavouring modi-
fication. The maximm incorporation would be 2 - 3%
by this
route so it appears unlikely to be an attractive
alternative.
b) Use of tobacco extracts
The limitations to the use of tobacco extracts are
the
quantities which can be applied and the increase
in weight
without a parallel increase in filling power. The
tobacco from
which the extract is obtained then represents an
additional
cost unless it can be utilised elsewhere. Unless
tobacco ex-
tracts are further purified to enrich their
nicotine content,
the latter will be present in the extract to the
same concen-
tration as in the original leaf (maximum
concentration to 502
solids).
In theory at least, the extract could be added as
a
casing or by DOQ application to the cut tobacco. A
third
alternative would be to use a high nicotine
extract (e.g. from
N.rustica) to supplement or in place of the
extract from waste
tobacco in sheet manufacture. All of these routes
would imply
additional costs from the tobacco to be used, its
extraction
and concentration unless sufficient quantities of
suitable
strength could be produced from waste or as
by-products of
other processes -Hag, DIET or Tamag for example.
Blend dis-
tortion could also occur, particularly in relation
to smoke
analyses from the addition of extra nitrate etc.
in the extract.
In the U.K., solutions containing 7.5% or more
nicotine would
be covered by the Poisons Act, so that
realistically the upper
limit of nicotine content for application might
be5Z. However,
this strength of solution would certainly produce
a major
increase in nicotine applied to the blend at 10%
(2% increased
to 2.5%).
c) Use of pure nicotine
Apart from legal and workplace restrictions,
nicotine
solutions offer a wide range'of opportunities to
fortify the
nicotine content of blends. Because nicotine
itself becomes
C>
NJ
CD
CO
BATCo document for Province of British Columbia 4
November 1999


-7-
oxidised on exposure to air, in practice the more
stable
salts would probably have to be used. Strong
solutions of
the salts are very viscous, but this is less of a
limitation
than the practical question of applying nicotine
at all
other than as an extract of tobacco. The Poisons
Act in..the
U.K. covers the use of nicotine solutions of 7.5%
or more. In
addition to the question of preparing any solution
for use by
dilution of the concentrate, in which 0.2 ml of a
302 concen-
trate would represent the calculated lethal dose
for a man
by skin absorption, the work environment would
have to be mon-
itored to ensure the air concentration did not
exceed 0.5 mg
nicotine per cubic metre. This strongly suggests
that the use
of pure nicotine or its salts would not be a
realistic prop-
osition in normal tobacco manufacturing operations
even if it
were acceptable to the work-force. This is
supported by the
report that addition of nicotine solutions to
sheet was tried
at Le Mans and required evacuation of the plant.
An alternative approach to the use of nicotine
would be
its addition in the form of micro-capsules, which
would minimise
the risk of any hazard to the work-force. This
method has been
investigated by Amcel as a way of adding nicotine
to Cytrel
and also by Imperial Tobacco (U.K.). In the latter
case, a
report pointed out that the use of nicotine in
this form would
cause the addition of an equal amount of wax. Thus
at the
present time it may be technically feasible, but
the effect on
the properties of the smoke are unknown.
Addition of nicotine to cigarette paper has been
con-
sidered in the past, but appears to offer no
advantage over
other approaches.
d) Augmentation of Smoke Nicotine
Although not immediately exploitable, methods
which
would increase the effect of the nicotine already
present in
the blend would have obvious attractions.
Few direct comparisons exist on the transfer of
nico-
CO
tine to smoke from different blends in the same
cigarette CN
BATCo document for Province of British Columbia 4
November 1999


-8-
design. Such information is available for a BAT
and a
Philip Morris brand from Ve-nezuela: nicotine
transfer was
reported as 8.5% for the former and 12.9% in the
latter, or
equivalent to a 50% augmentation of nicotine for
the Philip
Morris brandl
Increasing the free nicotine content in the smoke
by
making it more alkaline may be another approach or
directly
related to the problem of maximising transfer
efficiency.
Either way, whilst potentially very useful, major
changes in
smoke subjective character might be expected to
occur also.
3. ACTIVITY BY COMPETITORS
Information on this subject is largely anecdotal
and
there is remarkably little incontrovertible
evidence of spe-
cific commercial exploitation of nicotine
fortification of
blends. The notes which follow are illustrative
rather than
necessarily exhaustive.
a) American Celanese
Cytrel patents included examples involving the
addition
of pure nicotine to the formula. They appear to
have been well
aware of the various difficulties by also offering
micro-
encapsulated nicotine to fortify blends containing
Cytrel.
b) Gallahers
Reported to have approached Tamag on possibility
of
obtaining tonnage quantities of nicotine.
Apparently use
N.rustica in sheet manufactured for them at Le
Mans. Have
recently patented a cell culture process for
obtaining nicotine,
but this is not considered to offer a realistic
route to
commercial quantities of nicotine in the
foreseeable future.
c) Imperial Tobacco W.K.)/I.C.I.
Earlier activities were concerned with obtaining
nicotine
from waste tobacco as insecticide (British
Nicotine Extraction C:)
Co. Ltd.) and they developed an extraction process
at Limbe, C=)
o-,
(-rl I
Malawi. Thus they have considerable expertise in
the produc-
tion and part purification of nicotine extracts
from tobaccc
co
BATCo document for Province of BritiSh Columbia 4
November 1999


-9-
ources. Reported activities in the early '70's
were
:
hought to be wholly concirned with possible
application to
substitutes and included: an investigation of
micro-
encapsulation of nicotine (reported 1972); growing
N.rustica
in this country at about the same time an the
B.A.T. U.K.
leaf project and patent claims by I.C.I. for
addition of
N.rustica extracts to substitutes as flavour
materiall
d) Japan Tobacco & Salt Monopoly
Numerous scientific investigations on aspects of
tobacco
cell culture have been reported and some work has
been done
on a small pilot scale. Their original aim was not
nicotine
production, but to produce a tobacco material in
this way.
In this respect they made good progress and then
turned some
attention to improving nicotine yields. Difficult
to believe
it is a practicable alternative to field
cultivation at this
time.
e) Reynolds
Over 10 years ago they patented a process for
obtaining
high yields of hydroxynico tine, but it is not
known whether
this has ever been utilised. Opinion was that it
was intended
to be a way of stabilising nicotine in sheet to
obtain higher
transfer subsequently to main stream smoke.
It was reported that they had requested the
Variety
Trials Committee in the U.S.A. to modify the
standard nicotine
content requireddownwards, which does not suggest
a concern
to increase nicotine levels.
f) Tamag
Claimed that they have been approached by
companies such
as Imperial and Gallaher for tonnage quantities of
nicotine.
4. RESTRICTIONS ON THE USE OF NICOTINE
Nicotine is widely known as a poisonous substance
by skin
absorption, inhalation or ingestion. As an
example, its use
in the U.K. is controlled by the regulations on
Poisons
(Statutory Instrument 78, No. 1) which covers
solutions con-
C)
c0
co
BATCO document for Province of British COIUMbia 4
NoVernber 1999


_10-
taining 7.5Z or more nicotine either 88 the free
base or in
the form of any of its 'salts. Accordingly any
factory using
nicotine solutions, which would include the
application of
more dilute solutions prepared from concentrates
on the premises,
would need to meet very.stringent control measures
on storage.
measurement and use. In addition, the Health and
Safety
Executive would almost certainly require the uost
stringent
precautions in the handling, use and application
of nicotine
solutions and the strictest supervision and
monitoring of the
work environment. The strongest tobacco extracts
could just
fall within the provisions of the Poisons Act.
5. TOXICOLOGICAL ASPECTS
Apart from the acute toxic hazards which are
mainly the
concern of the regulatory aspects (4 above), there
has been
extensive investigation of the involvement and
significance of
nicotine in any toxic properties of smoke. The
current position
can be summarised as follows:
a) it is not itself carcinogenic;
b) There is no conclusive evidence of teratogenic
action;.
c) It is probably not involved in the pathogenesis
of athero-
sclerosis, but may be significant in relation to
the
increased risk of smokers for myocardial
infarction and
coronary disease;
d) In tobacco it is a precursor of the
carcinogenic N-nitro-
sonornicotine;
e) it may have a role in the aetiology of
carcinoma of the
.pancreas;
Q It may exert a co-carcinogenic effect in tobacco
smoke
under experimental conditions.
From all of these aspects, the most immediately
relevant
appears to be the question of its co-carcinogenic
action in
tobacco smoke condensate. The early evidence on
this subject C=)
was often conflicting, with some studies showing a
reduction CD
in tumorigenicity by the application of nicotine
tartrate
as well as the carcinogen benzpyrene compared with
the latter
alone, or no effect on the tumorigenicity of a
smoke fraction c0
BATCo document for Province of BritiSh Columbia 4
November 1999


_11-
fron the addition of nicotine tartrate (TRC
studies). An
experiment to clarify ;he position (Jan., 314) by
adding extra
nicotine to a low nicotine bland has shown
evidence of an
increased "suspected" tumoriSenic ratio with
increasing nico-
tine level..
The results of this study have to be confirmed by
further
analysis and the range of nicotine dose was very
large: from
0.56 - 1.62 mg TNA.
Nicotine is considered to be co-carcinogenic by
one of the
U.S. experts, Dr. F.G. Bock. Using pure chemicals
and combining
the application of a carcinogen with a promotor
and with nico-
tine, low levels of the latter enchanced
tumorigenicity, but
high levels inhibited it. An experiment is now in
progress in
the U.S.A. which is closely comparable with B14
using varying
levels of added nicotine salts to both flue-cured
and Burley
tobaccos, mainly in the form of SRT. It must be
assumed as
likely that results similar to the B14 study will
be obtained.
Some further confirmation of higher specific
activity
of condensate from the addition of nicotine to the
cigarette
has come from testing the B14 series for mutagenic
activity in
Canada, the results closely paralleling the mouse
skin study:
Cigarette TNS(mg) Preliminary Results Ratio of
(suspected carcinomas) Mutagenic Activity
:Tumorigenic Ratio
B14/1 0.56 1.0 1.0
B14/2 0.8 1.2 1.04
B14/3 1.22 1.28 1.22
B14/4 1.62 1.41 1.44
Players Check 20 1.70 0.98
0-,
(-n._-!,
The results for the 'Players Check 20' suggest
that . ....
activity is not related ibsolutely to nicotine
level, but more
BATCo document for Province of BritiSh Columbia 4
November 1999


information is required to assess its
significance.
6. CONCLUSIONS
At present it would appear inadvisable to propose
add ing
nicotine alone to cigarettes both because of the
practical
difficulties of doing so and the preliminary
evidence of
increased specific activity of wioke condensate
associated
with its addition.
The simplest route to nicotine fortification of
blends
immediately available is from the selection of
high nicotine
tobaccos and this may represent the cheapest route
also if
it is to be done on a large scale. It is within
normal and
traditional blending practice even through there
may be
limitations on the degree of fortification which
can be
obtained. The effect on smoke condensate specifici
activity
is uncertain, but it should be appreciated that
even this
route may cause an increase.
If the preliminary results of the B14 experiment
are con-
firmed and similar ones obtained in the U.S.A.
experiment,
pressure for smoke nicotine levels to be lowered
would mount.
The longer term solution may well lie in the
augmentation of
the transfer and impact of the smoke nicotine,
rather than
fortification of the blend itself.
Ak
T. G. MITCHELL
CrN j
BATCo document for Province of BritiSh Columbia 4
November 1999