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" :~i~if!!<'!:<;::il~ :( BRITISH AMERICAN TOBACCO Copy Request Form Request Number: Organization: Physicians for a smoke free Canada 8 Request Details Request Date: ' oe • a • • o•olooeoeue • • le •o •me • • •• •a • • • ••••cool•I•••.• File Number: /~ ~] ..~..~. ~ 6.Box Number: ...... .~7..!.~..~.. .................... Page Range: First Page Last Page //. 0 ~ ~ / ~" ~-- / Requested By: (Print Name) Details below will be filled in by Depository Staff Only II Copy Details Copied By: .... i~ ................................ Date: .... .~../S.¢~.~./.~..~.. ............. Time:. ~.'~.#~. .............. Copy Checked By: • • • • •Q• • • • eeoooeolooooeeooo•eeeoe • eeooe•o o• coot,••co o•o•o • • Date: ..... Time: .......... e•• • • ee•e e•e•o • • • oe rio•e••••••• e•• •••oo • • •o•0oo••o•• III Delivery Details Checked By: ............................ ........ Date: ................... Sent By: ........ Date: ..... . ........... ooe•eooo• • .el • • ••ooo•••••o•Q•lo••o o•• • BATCo document for PFSFC 1 March 1999

-q VIr ' i ! _ ° Imperial Tobacco Limited/Limit~e RESEARCH AND DEVELOPMENT DIVISION MONTREAL THIS REPORT MUST NOT BE COPIED OR SHOWN TO UNAUTHORIZED PERSONS. O L~ Co BATCo document for PFSFC 1 March 1999

COPY NO. ~fiDgR~J~L T~BACCO LTD. HOWI'1~),L R.ES'I'R ZC'I'~D PROGRESS I~PORT JULY 1993 - DEC'-I~LBER 1993 B.A.T. CONFIDENTIAL - CATEGORY I: MINNESOTA TOBACCO LITIGATION BATCo document for PFSFC 1 March 1999

- I - L~.OI~Ry PRODUCT RESEARCH O B.A.T. CONFIDENTIAL - CATEGORY I: MINNESOTA TOBACCO LITIGATION BATCo document for PFSFC 1 March 1999

RECORD T~PE: SUB TYPE: SECURITY CODE : FUNDING BODY: ORGANIZATION: GROUP NUMBER : LOCAL PROJECT ~r~,,J3-:~(S): PROJECT TITLE : PERSON RESPONSIBLE: EFFORT: PROJECT DESCRIPTION: S COPE: DEPTH: FUNCTION: OBJECTIVE: CLUSTER: DATE REVIEW WRITTEN: REVIEW TITLE: REVIEW TEXT: - 2 - P S ITL CANADA 501 T-7281 ;T-7099 ;T-6967 Visa YOUSSEF, M. 0.i * 1993 The goal of VZSA is to investigate produc~ oppo~unities which address the social and legislative restrictions that ",...he smoker faces on she use of cigarettes. Focus will continue on tasting new papers as received from suppliers and exploring recipe modifications (T-?099}. This area also involves keeping a watching brief on ETS studies conduc1:ed elsewhere (T-7281). GROUP BENEFIT FUNDAMENTAL GENERAL ALTERNATIVE PRODUCT RES/DEV January 1994 Visa Since the June 1993 Progress Report, a package panel test was conducted for the latest low eLdest=sam prototyl:e #827, made to du Maurier Light K.S. (duMLKS) specifications with Ecueta's sldeiureampaper TaD 06651 (elect=ape=rotated to 30 CORESTA with added flavouEs}, Ratio falters and recipe C01. Sample #827 was compared to previous sample #321, made to the same specifications, but with Ecusua TaD 06650 paper (8 CORESTA - with added flavours) and recipe C26. Based on the resul~s, sample #827 was found no be slightly stronger with marginally more tasT.a and irritaulon. It also had more impact. Appearance and burning properties were similar to sample #321 (whitish ash), however, the ash appearance of sample #827 is close= to the target cigarette (du MLXS). Overall, sample #827 appears to be the best low sidestrlam prototype to date. Results were presented to Marketing. A focus group may be conducted in Spring of '94 for Sample #827 to reassess consumers interest in a low sidistre~ product. CD B.A.T. CONFIDENTIAL - CATEGORY I: ~IINNESOTA TOBACCO LITIGATION BATCo document for PFSFC 1 March 1999

-3 - RECORD TYPE= SUB TYPE= SZCURITT CODE: FENDING BODY: ORGANIZATION: GROOP MEMBER: LOCAL pROTECT NUMBER ( S ) : PROJECT TITLE : PERSON RESPONSIBLE: EFFORT: PROJECT DESCRIPTION: SCOPE: DEPTH: FUNCTZON: OBJECTIVE: CLUSTER: DATE REVZZW WRITTEN: REVZEW TITLE: REVIZW TEXT: a) ZTL CANADA 578 T-6431;T-7711-2;T-6542;T-7344;T-6968;T-5549 Day PORTER, A. S.7 t 1993 This projec~ involves the development of a tobacco combustion cigarette of conventional appearance but which produces smoke with low specific mutaginicity and low concentrations of PAH, phenols, vapour phase aldehydes, HCN and CO. This product will be designed to provide acceptable taste and nicotine, while producing reduced levels of 8idsstresm smoke compared with conventional produc~e. The principal strategy for achieving reduction of -...he paniculate smoke noxae is by ~obac¢o modification using solvent extraction (T-6431), enzyme/microbial treatment (T-7711-2}, and reconstitution (T-6542). Emphasis will also he placed on retaining tobacco flavour components during the various modification processes. Vapour phase texas will be reduced by conventional cigarette components such as absorben= filters and ventilation (T-7344). Analytical method development specifically for DAY is also included in this project (T-6968, T-5549). Successful developments of low sides=ream products via proJsc~ VZSA will he incorpora=ed into DAY as appropriate. GROUP BENEFZT FUNDAMENTAL GENERAL ALTERNATIVE PRODUCT RES/DEV January 1994 Day Tobacco Frac~ionation ~De Gr~dp~ Y.) (T-6431) ~b~ec~£ve Evaluation of Treated Samples= A formal evaluation of the l@jec~ive ~king properties of cigarette #645 (process described in last Progress Repo~c) was made by the ZTL Expert Panel. In general this sample had a very poor smoke with below average mechanics, low impact, low level of tobacco taste and unacceptable irritation. The tobacco taste was flat with an unpleasant non-tobacco taste. A new sample was prepared using the technique described in the previous Progress Repor~ (Jan. '93 - July '93). In order to improve the ~king properties of this cigarette, the solubles level was increased from 45% (#645) to 60% of the total tobacco weight. Cigarette #719 was made using ~he 60% W/W solubles treated tobacco using DM~S construction parameters. Another sample, cigarette #518, was 0 B.A.T. CONFIDENTIAL - CATEGORY I: MINNESOTA TOBACCO LITIGATION BATCo document for PFSFC 1 March 1999

- 4 - prepared with water only extracted V-09 with the concentrated Jolublmm added back without any treatment. The tobacco used for cigaEsCts #418 was thus processed in the same way as samples #645 and #719 but no specific tobacco coE~onsnts were removed. I~ was used as a conceal tobacco. Tobacco and smoke chemistry foe cigarettes #719 and #518 are prelenced in Table #1 toqecher w£th data for o£qa~s=~es #837 (un~=e&ted V-09) and #645. TABLX 1: PETSZCAL AND SMOKING RESULTS Leaf Nic. (q) Red. Sugars ( • ) CAge. P.D. It) Puff Number (~/ci;C) (.~/puff) TAR (.~/cl~) l~l~f~) (.~Iclcjt) (rag/puff) (mg/ci~) (mg/puff) CO (mg/cig~ } {mg/puff) COlT T:NrC. #837 2.67 21.2 121 10.0 23.02 2.30 I?.41 #645 2.03 18.2 123 9.4 18.42 1.49 14.02 #518 2.21 20.2 106 9.8 23.81 2.43 18.82 1.74 3.12 0.31 2.49 0.25 18.I5 1.82 1.04 7:1 1.49 2.73 0.29 1.67 0.18 15.02 1.60 1.07 8:1 1.92 3.28 0.33 1.71 0.17 17.48 1.78 0.93 11:0 #719 2.43 23.6 106 9.8 19.89 2.03 15.37 1.57 2.68 0.27 1.84 0.19 15.96 1.63 1.04 8:4 High nicotine recoveries were obtained for cigarettes #518 and #719, and ~hs tar to nicotine radios era good. Cigarettes #518 and #719 were smoked for phenols {hycLToquinone, resorcinol, cateohol, phenol and o-m-p- cresol), aldehydes (formaldehyde, acetaldehyde and acrolein), benzo(a)pyrene, HCN and Ames specif£c activity. The results are reported in Table 2. CD m --d B.A.T. CONFIDENTIAL - CATEGORY I: MINNESOTA TOBACCO LITIGATION BATCo document for PFSFC 1 March 1999

- 5 - TAS~.~ 2, S~0KZ ~T.yS;S Smoke Co m~onenua Puff Numbec TAR (mg/cig~.) P"~NOLS ( ~g/c igt ) Ratio (#g phenolslmg tax') xu~zs (.glc.-' ~) #837 i 10.0 17.4 324 19 1461 #64S 9.4 14.0 152 11 1672 #51S 9.8 18.8 235 13 1530 #719 9.8 15.4 180 12 1652 Ratio (#g aldehydes/ms tar) B(a)P (ng/cig~) Ratio (ng B(a)P/mg tar) HCN (pg/cig~) Ra~io (gg HCN/mg tar) Spec. Ac~ivlty 84 19 1.1 295 17 1.3 120 15 1.1 102 13 0.53 81 12 0.7 317 17 1.2 10? 14 0.9 164 11 0.63 B) The deliveries of the two fully Creaced cigarettes #645 and #719 are lower than the two control clgatette8 (#837 and #518) except for aldehyde deliveries (see previous Progress Repo~). Subjectively cigarette #719 was an improvement Over cigarette #64S. However it m:ill had a low impact, I=e ~:Itanion, • modifi~ tobacco flirt an a slight off-Baste despite Bigh tac and nicotine deliveries. Modification of the pH of the aqueous extra¢~ (from 5.5 to 8.0}, as suggested in =he last ProgEes8 Report, had little effec-, on the Impac~ of ciga~s~.~es #645 and #719. We are presmn~!y evaluanlng Bhe e~fsc= of Bhe extraction procedure on the tobacco taste level and on the impac~ of ~he i~oke. These investigations will be done with a new control blend since the taste level and impa~ of the untreated control (#837) was surprisingly low. $ub~ec~ive evaluation of the new control (V-23, lamina blend of JPSI showed high taste level and high impact.. AldehYde R~uc~ion: ABtemp~e to reduce aldehyde deliverils of #645 were made using absorbenB fil~ers: carbon, ductile, carbon + duolite, Hg 8illcaue ÷ carbon and sodium ¢aEbonate. Carbon was used in a triple falter configuration at 75 ms/tip. Duolite It 60 ~/tip was in a dual dalmatian conflguraClon; carbon + duolite (75 ml and 60 mq respectively) was in a triple configuration; M~ silicate ÷ ~arbon (?S m~) was in a ~riple. Sodium carbonate was loaded into preformed cellulose acetate filners from solution to give 25 ms/tip. All ~ilters were hand attached ~o #645 rods. P.O.'s ~eEe ± 20 ~=u of the contro~ (C.A.). Aldehyde deliveries ws~s: CD N B.A.T. CONFIDENTIAL - CATEGORY I: ~IIN~ESOTA TOBACCO LITIGATION BATCo document for PFSFC 1 March 1999

- 6 - Cellulose Acenaue (Ccn~rol) Cazbon Duolite Carbon ÷ Duolite Mg silicon8 + Carbon Sodium Carbonane TQ r • Ace, Act. (ug/cig. 242 11S (52) 73 (70) 50 (79) 144 (40} 115 (53) (q Reduction va 1250 10SS (15) 1195 (4) 869 (31) 1044 (17) 1111 (11) Control} 180 S4 (?0) 114 (37) 38 (78) ~l (6o) 175 (3) Results wore as expecned exceF for the mll effect of duellos on acetaldehyde. This m87 be due to aging of the duellos (6 yrs.), which also gave a pronounced off-taste. Sodium carbonate was effective for fo~aldehyde buu noC the ocher aldehydes. Future work will include evaluation of DLaion Resin repoECmd under ?344. ~-~CALE PREPARATTON OF RECONSTTTUTED TOBACCO SHEETS (T-6542) s0H~A~: Reconstituted tobacco sheens were made using a laboranory drum casner and the following componenns: ~obacco flour (83-89% W/W), sodium carbox~hylcellulose (5-10% W/W}, glycerol (5% w/w) and various salts as addinives (1-2% W/W). The following additives were used in the hope of improving the sheen flavour: ammonium phosphanemonobaeic, (NH4)g~PO,; ammonium blcarbonate, (HN,):C03; 1~Ycaesium carbonate, K~O3; sodium carbonate, Na~; and ammonium citrate dibasic, C~H~.~OT. The sheens will be shredded and puc in duMl~S ¢igare~¢e tubes for subjective evaluation. J~ CD P~ J~ q B.A.T. CONFIDENTIAL - CATEGORY I: ~IINNESOTA TOBACCO LITIGATION BATCo document for PFSFC 1 March 1999

- 7 - RECORD TYPE : SUB TYPE: SECURITY CODE: FUNDING BODY: ORGANIZATION : GROUP HUMBER: LOCAL PROJECT NUMBER (S) : PROJECT TITLE = PERSON RESPONSIBLE: EFFORT: PROJECT DESCRIPTION: SCOPE: DEPTH: FUNCTION: OBJECTIVE : CLUSTER: DATE REV~.EW WRITTEN: REVIZW TITLE; REVIEW TEXT: Presence of Oraanic Solvents: P $ ITL CANADA 416 T-7711-2 Enzyme/Microbial TreaUmenn of Tobacco BILIMORIA, M.H. 0.8"1993 This s=udy focuses on uhe hydrolysis and removal of ~obacco polymeric materials such as pro~eins, pectins, e~o. and ~heir d~rada~ion produc~8 using enzymaa and/or microorganisms. GROUP RELEVANT FUNDAMENTAL GENERAL ALTERNATIVE PRODUCT RES/DEV January 1994 Enzyme~Microbial Trea~menn of Tobacco i. Enzymatic Protein Removal from Tobac~q ~,n Some enzymes, such am llpames and peeresses, ~a known =o bo more s~ablo in organic 8clven=s ~han in wares. Consequently have dote~nined ~he pEo~eoly~ic ac~ivi=y of ~he Asoeraillua pro~eaee (Si~na P-4032) in ~hree solvents, vlz, acetone, euhanol and me~hano! in concentrations ranging f~ 10t ~o 50%. PROTEZN REMOVAL FROM FLUE-CURED TOBACCO BY ASPEROZLLUS ORYZAE PROTEASE IN TEE PRESENCE OF ORG~ZC SOLVI~NT5 SOLVENT W~TZR 10t 20% 50% ZT~A~OL 10% 20% S0% 10t 20% 50% t BSA as s~andard. PROTEIN CONTENT - MG/GM DWRT TREATMENT CONTROL - NO PROTEASE 68.9 68.1 66.4 59.7 (13) 67.2 70.4 66.8 69.4 71.1 62.1 (I0) PROTEASE 29.1 (58) 30.8 (ss) 38.1 (45) 52.8 (23) 37.7 (4s) 48.8 (29) 60.6 (12) 36.g (46) 39.9 (42) 59.0 (14) The samples were treated with pro:ease (5 mg/gm tobacco) for 18 hours on a rotary shaker at R.T. The numbers in bracke~8 are per csnn reductions as compared to aqueous pro%ease ~rea~men~. CD P~3 B.A.T. CONFIDENTIAL - CATEGORY I: MINNESOTA TOBACCO LITIGATION BATCo document for PFSFC 1 March 1999

- 8 - F=om the =esu1~.o of this study, ~: wi%l be seen that 10% acol:one did not affect protein removal from flue-cured tobacco (55% =amoral vs 58t for the water control). At hlgheE acetone concentEationa smaller amounts of tobacco protein were oolubilized. ~hanol and ~thanol appeared to be more Inhib£uo:y than ecrcone, giving signlfi=antly smaller protein reductions even a~ I0% concentration. The experiment web repeated at 5% and 7.5% solvent concentrations when once again acetone dad no~ show any Inhlh£tory ac~Ivlty, while ethanol and methanol r~uced ~oteol~ic a~ivi~y even au Uhoee low ¢:ncont:ations. TX~2 PROTEX~ REMOVAL FROM FLUE-CURED TOBACCO BT ASPERGI'LLUS ORY~AE PROTEASE ~/~' Tn FRZS~(:Z Or O~OAN~C SOLVm~ III ENZYME TREATMENT IN PRESZNCE OF PROTEIN CONTENT - MG/GM DWB. t WATER St ACETONE 7.5t ACETONE 5q ETHANOL 7.5 t ETHANOL $q ~ETHANOL 7.5t M~THANOL CONTROL - 7.5• ACETONE CONTROL - WATER tIcJU as ~canda_--ct. 33.6 (63) 32.4 (64) 34.9 (61) 43.9 (52) 39.9 (56) 39.8 (56) 41.s (54) 88.3 90.6 I The samples were treated wi~h pEocaaoe (S mg/gm tobacco) for 18 hours on a ro~a~ share at R.T. The numbers in brackets are per cent reductions ae compared ta ~he water con~Eol. 2. ~nzvmatic Protein Removal from Differen~ Tobacco Tv~es: When flue-cured :obac:o ks treated w£th ~yperaillus protease (P-4032), more than 60• of ~s prote£n Ls solubillzed. Whether a similar solubillzation w£11 OCCUE when other tobacco ~ypeo are sim£1anly ~Eeatad has :econtl7 been tried. B.A.T. CONFID~-NTIAL - CATEGORY I: MINNESOTA TOBACCO LITIGATION G ~o "-,l BATCo document for PFSFC 1 March 1999

-9 - 3. PROTEIN SOLD~ILIZATTON ~N DZFFERENT TOBACCO TTP~q TOBACCO TYPE AND TREATMENT TIME HOURS 2 4 6 2 4 6 2 4 6 t I9~ am standard. PROTEIN CONTENT - MG/GM D~,'B7 CONTROL 82.6 B3.0 83.5 122.3 124.1 120.6 175.3 17S.3 177.4 ENZYME-TREATED 49.1 (41) 44.4 (47} 44.9 (46) 98.1 (20) 80.1 (36) 80.5 (33) 182.1 168.5 (4) 164.0 (8} The aamplee wmre treated with proteaae (S mg/gm tobacco) for up co 6 hours in an environmental shaker eC 50"C. The numbers in brackets are per cent reductions as compared to water controls. From this table At will be seen that while treatment of tobaccos wiT, h Aeveraill~ proteaee removes about 4Sq of the protein in flue-cured tobacco within 4-6 hours at S0eC, only about 35% of the protein in oriental tobacco is solubillzmd. The protein in the dark tobacco resisted soluhilization under the above conditions. More protease per gram ~oba¢¢o and longer incubation may be needed to release the tightly bound protein in this highly oxidized and changed tobacco. Proteolv~ic Activity of Some Yeasts ~solated trom Tobacco: Many of the yea~e we have isolated from tobacco can hydrolyze proteinl such as casein and gelatin when these ere incorporated into agar media and some of =hem can even arrack protean complexes such as those An ~annic acid-brain heaE~ infusion agar medium. However, they have failed to hydrclyze oE solubillze protein in flue-cured tobacco. Consequently, before unde~-~aking any fu~her studies on protein removal from tobacco by yeast ~rea~ment, we decided to screen several of our proceollr~io yeasts for their ability ~o eecreUe proteaeos in liquid media. In order to measure exocellular proposes we have adopted a e~mple and highly sensitive colorimetrio procedure involving a dye labelled protein substrata (hide powder azure). When this protein substrata £e hydrolysed by • proteaee, soluble dye-labelled peptidee and amino acids are released into the reaction mixture and are then measured colorimetrically. For measuring exocellular enzyme production by our ~obacco yeasts, supernauants from cultures grown in liquid media containing bovine serum albumin (BSA) or casein have been used. The protein content of ~hmse culture superna~anns has also been determined, as another measure of yaae~ prcceolytic activity. The results of such a screening are presented in Table 4. CD t~ CC B.A.T. CONFIDENTIAL - CATEGORY I: iA'IINNESOTA TOBACCO LITIGATION BATCo document for PFSFC 1 March 1999

- 10 - PROTEOLTTZC ACTI"V~TT 0F SOM~t TEAST$ ISOLATED PROM TOIL).CCn PROTEI2~ CONTENT OF 7-DA~ PROTEASE ACTIVITY IN 7-DAY ~EAST SUPERNATANTS -mg ml~ SUPERNATANTS - U/el* Medium A Medium B Medium A Medium B Con=rol 2.35 2.2S Hansenula anomala 0.03 (99} 0.04 (98) 0.437 0.428 (0-0) RhodoCorula glucinis 1.90 (19) - Nil Nil (3-9) Uniden=ified Yeast 1.90 (19) 2.00 (11) Nil Nil (165-0) Rhodocorula glutinis 2.00 (15) 2.25 (0) Nil Nil (407-0) candida Caselnoly~ica 2.20 (6) 2.10 (6) Nil Nil GglS062B Medium A: BSA 0.2%; Glucose 1.0q; YNS (no a.a., (NH,]~O,) 0.17%. Medium B: Medium A + Tweet 80 0.05%. " A unln of pEonease activity is than amounn of enzyme which gives an increase of 10.D./min/ml a~ 37eC and pH 3.0. The numbers in brackets ace per cent decreases from conurol. Only Hansenula anomala secreted measurable amounts of an acid pconease under the conditions of the experiment. DeteEminatlon of the protein conusnt of the supeEnatant also shows significant pronaoly1:ic activity only in this Hansenula culture. The culuurs of Cand~da caseinolv~ica, kindly provided by Dr. M.A. Lachancs of the University of Western Ontario, London, known ~o be highly proneoly~ic, also failed to utilize BSA or secrete prc~eoly~i¢ enzymes under the condit£ons of the experiment. Next, we proceeded to determine the rate at which pronsass is secreted by ~ansenula ancmala. From the time study, presenusd in Table 5, An will be seen thau there is significant pronease produc=Ion within 24 hours of growth ~ the BSA-con~aining media, and than maximal enzyme secretion occurs within 3-4 days, and possibly earlier in the nween-containing medium. r,~ u m B.A.T. CONFIDENTIAL - CATEGORY I; MINNESOTA TOBACCO LITIGATION BATCo document for PFSFC 1 March 1999

- 11 - PROTEOLYT~C ACT.~VITT or ~A~S~ A~rOMAT,.A PROTEIN CONTENT OF SUPERNATANTS -mg ml"~ PROTEASE ACTIVITY OF SUPERNATANTS - UNITS/ml* CULTURE FILTRATE AT Medium A Midium g Medium A Medium B 24 hrs 0.04 (98) 0.0S (98) 0.40S 0.479 48 h=s 0,04 (98) 0.02 (99) 0.479 0.$90 72 hrs 0.03 (99) 96 hrs 0.04 (98) 7 days 0.03 (99) 0.04 (98) Con~=ol 2.35 2.25 Medium A: BSA 0.2t; Clucos8 1.0%; YNB (no as, Medium B: Medium A + Tween 80 0.05% 0.512 0.580 0.523 0.531 0.378 0.361 Nil (~,)-.so,) 0.17t • A uni~ of pro~ease a~ivi~y Lm ~ha¢ amoun~ of enzyme which results tn an increase of 10.D./minute/ml st 37"C and pH 3.0. The numbers in brackets are per cent decreases from control. The inability of Candida caseinolv~ica and onhor yeasts to secrete pEo~oasm tn BSA-con~aining media, suggesns a specificity in ~e=ms of protein EequiEemen~ for enzyme induction. Since Candida case£nol~ica ks known ~o secrete p~otease on caeein-con~ainLng medium, we decided to soften some of the yeasts on such s medium. PROTEIN DECOMPOS~0M BY YEASTS OROWW ~ RSA- AND C.AS~N-COt4TA;NTNG ~ZDZ& YEAST Haneenula Anomala (0-0) Cryp=ococcus Al~idus _ (58-0) Rhodoto=ula Glucinis (92-PI) PRDTEIN IN ~IUM BSA Casein BSA Casein SSA Casein Candida BSA Caseinolyuica Casein GgI5062B con~:=ol BSA Casein PROTEIN CONTENT OF CULTURE SUPERNA2ANTS INCUBATION PERIOD - DAIS m, 0 1 2.04 2.00 0.64 Nil 2.00 1.92 0.70 0.0S 2.08 2.08 0.42 Nil 2.06 2.04 0.76 0.47 2 3 4 1.96 0.12 0.045 Nil Nil Nil m, 1.98 2.04 1.92 0.02 Nil N~I 2.06 1.76 1.42 Nil Nil Nil 2.06 2.16 2.10 0.52 0.52 0.51 2.00 2.04 2.06 2.10 2.04 0.88 0.65 0.66 0.62 0.61 HIll 7 0.045 N£1 1.S8 Nil 0.79 Nil 2.08 0.43 2.10 0.64 III CD CO CD B.A.T. CONFIDENTIAL - CATEGORY I: ~IINNESOTA TOBACCO LITIGATION BATCo document for PFSFC 1 March 1999

- 12 - P~0TEXSZ ACTZVTTT OF TEXS'CS ~¢W~r ~N S$~- A." CXS~:N-CONTA:NZNa ~EDZA YEAST PROIT%N IN MEDIUM PltOI'F.ISE ACTZV~TTt OF SUP~ - UN~S* /NL INCUBATZON PERIOD - DAYS 0 1 2 3 4 7 Nil Nil 0.015 0.212 0.214 0.198 ~il 0.190 0.196 0.248 0.320 0.236 Hansenula BSA Anomala Casein (0-0) CrylOCOCOC~4s BSA Nil 0.044 0.004 Albidua Casein Nil ~il Nil (58-0) RhodoCocula BSA Nil Nil Nil Glucinis Casein Nil Nil 0.016 (92-P1) Candida BSA ~il 0.006 Nil Cameinoly~ica Casein Nil Nil 0.022 GglS052B t De~ermined using Hide Pow~sr Azure as su~s~Eaue. " 1 Unit - 10D. change per men. per ml a= 37eC. Nil 0.011 0.010 0.031 Nil Nil Nil 0.02g 0.016 0.032 Nil Nil Nil Nil Nil 0.014 0.008 Nil F=om the results of this ex3~r~nenC, it will be seen thac casein appea~rs =o be more suitable chat BSA in re:ms of procease production by yeasts. This is clear when bo~h protein (Table 6) and enzyme (Table 7) are measured in the culture supernatants. It should be pointed ouC Chat the levels of protsase recorded in Hansenula yeast grown in BSA medium a=s lowmr than chose recorded in an ear!is= experiment {see Table 5), the difference being marked at the 24 and 48 hour gro~r~hs. This difference may be due to the procedure employed foe growing uhe yeas= inoculum. In the earlier experiment, the inoculum grown 30 hours with shaking at 30°C, was allowed to grow a further 2 days as a stationary culture aC ambient temperature (starvauion helps proCease production?). Further, the pro~ease assays were performed using a new batch of Hide Powder Azure (HPA) which gave high and erratic blanks. When contacted, the supplier admitted quality prohl~ and a new source of HPA is being sought. Finally, it should be noted chat our Hansenula yeast is a more prolific producer of pcocease Cha;n Candida caseinolv~Ica which is known for ice proCease secrecor/ activity. Of course, i= is also possible chau this Candida yeas~ needs a different medium for optimal enzyme secretion or that its procease has a dlfferen= optimum pH of ac=ivityand consequently, compares poorly ac pH 3.2, ~he optimum for the Nansenula pro=ease. Zf this Hansenula yeast is indeed a prolific producer of proCease, then its potential foe ~eparing this enzyme on an industrial scale also merits further investigation. 4. The Determination of P~ote£n in Plant Materials - Final Tab~e~: We have now co:pieCed our study on protein determination £n plant materials. The success of ~his procedure depends on ~he rapid extraction of protmin8 by ~iling 1 minute in 250-500 volumes of alkali, c~oling and then measuring protein in the extract, using the ~radford reagent. The concentrations of alkali solutions suitable for protein extraction from differenu plant materials are as follows: CD m co B.A.T. CONFIDENTIAL - CATEGORY I: MINNESOTA TOBACCO LITIGATION BATCo document for PFSFC 1 March 1999

- 13 - Fresh, leafy materials (including uncured, fresh tobacco) Cereal, legumes and other seeds Cured tobaccos F.-uits 0.2 M NaOH 0.2 M NaOH 2.0 M NaOH 2.0 M NaOH The resultm of this investigation will be w~itton s! a rtpor~. We have also measured the "pcotoin" con~ent of c£garecte s~:ke condenst¢s. 0.1-0.2 M NaOH was found to be superior to 2 M NaOH for extracting smoke protein. The rmmult8 are given Ln Table 8 below: CAULZ 8 PROTETR CONTENT 0F SMOKE CONDENSATES FRON FL~- AND ~J~K. &~R-CURED TO~%¢C~)$* Protein Concent~ of Condensate from (q) Protein Extraction in 200 Vo1~8 of Water TRIS-HCL Buffer, pH 7.5 0.1M 0.2 M 2.0M Pla~er'8 Check 28 Gsulo£see PEo~eln Extraction Without Boiling Nil 0.40 10.3 9.8 7.2 Boil£n~ 1 Minute 0.40 1.20 11.4 11.S 9.0 Without ~£11n9 Nil 0.1 12.4 11.8 7.0 m The c£qaEe~es were smoked using the K.R. Capillary Press Machine. f IgG as standard. Proteins=soul material was readily extracted in alkali at ambient temperature and boiling made llt~le dlffoEonce. Water and T=is-HCI buffer, pH 7.5 expiated little protein. Fu~he~, equal auaounts of pcotsin were extracted from flue-~ured and dark, air-cured tobacco c:ndansatee, even though the dark tobacco has ~wi¢s ~e amount of protein p=ssent as the flue-cu~ed tobaccQ. In both cases about 10q of the smoke condensate was decemined =o be p~o~ein. This appears to be cather high foe a pyrolysis produc~ llke smok~ condensate, and interference by phenolic compounds in the protein allay procedure cannot be ruled out. Studies to ¢onfizm ~he presence of such large a;uounts of protean in smoke condensable a~e in progress. Boilin 9 i Xinuco 0.80 1.0 12 • 6 12,8 5.8 CD PC I CC P~ B.A.T. CONFIDENTIAL - CATEGORY I: MINNESOTA TOBACCO LITIGATION BATCo document for PFSFC 1 March 1999

- 14 - I -- I= I RECORD TYPE: SUB TYPE: SECURITY CODE: FUNDING BODY: ORGANZ ZATZON: OROOP NUMBER: LOC~ PP.n~CT L~D4BER(S): PROJECT TITLE: PERSON RESPONSIBLE: EFFORT: PROJECT DESCRIPTION: SCOPE: DEPTH: FUNCTION: OBJECTIVE: CLUSTER: DATE REVIEW WRITTEN: REVIEW TZTLE: REVIEW TEXT: sez"v£cei have been rendered: So 2* 3. ITL CANADA 416 T-0111 Non Rouulne Analytical Service to Support: R & D Projects and for General Troubleshooting POULIN, p.; DUMONT, J. 1993 Work under this projec= i8 designed tO provide Analytical Suppo~ for req~es=s made by Purchasing, Technical Services, Manufacturing, Marke=ing and R & D. In addition any moni=oring uo Ia=isfy govecnmenB guidelines falls within Bhe scope of =his project. LOCAL SUPPORT GEN~ SUBJECTIVE METHOD DEV January 1994 NOn-RoutineAnaly~ioal Projects to Suppo~ R&D Projects and for General TroubleIhooting. Since r.he lasu rIview (July 1993) the following Spearmint Analysis on Cameo Spec~1: In septlm~er one (I) sample was analyzed fo= spearmint. The average level was 247 = I ppe which is outside the specification of 160-220 ppm. In November, two (2) sa~rples were analyzed. The average level was 202 ± 12 ppm which is within =he specification. 0ualitv Control of Menthol CrYstals for Guelph PlanP: Twenty-two (22} random samples of menthol crystals from 2 shipments have been analyzed since July 1993, and found to be within specifications. Coloration Problem of Menthol at Guelo~ P~anP: On Sip=ember 8, 1993, the yellow-beige colour of the menthol in the bath reappeared. The bath was emptied and cleaned. The nex~ day, before any production star~ed, the yellow-beige colour appeared again. On both days the menthol in the two feed tanks was clear. It was discovered that the solenoid controlling the menthol flow and the warming jacket around this solsnoid were bo~h leaking a black tar-llke subs=ante into the menthol bath. The solenoid was replaced with an insulated jacket type and a new warm£ng ~acket was placed eve= it. A small curved pipe was also installed to position the solenoid valve away from the Bop of the bath. Analysis performed on the bla=k tape of the old solenoid showed that the glue from this tape could have contributed =o the yellow-beige colour of the menthol. After a visit to the Guelph menthol room we concluded tha=: 1. The installation of the new solenoid valve controlling the =enthol feed should eliminate this source of conga=cite=ion. CD tJ~ C~ B.A.T. CONFIDENTIAL - CATEGORY I: MINNESOTA TOBACCO LITIGATION BATCo document for PFSFC 1 March 1999

- 15 - 4o 2, The source of the oil deposit on the Doctor Blade should be locaued and any Leak =epai=ed, and all ~he locations contaminated by o£1 and greases should he cleaned. 3. Dus~ foil accumulation in the men=hol bath should be minimized. Since it seemed to accumulate a~ the bottom of the banh At should not be a problem if production of GOLD foil stays low and if ~ha dus~ ks not contaminated by oil. 4. Until the end of January 1994, samples of menthol from the bath will be taken during each production day to monitor discolouration, if any and identify under wha~ clrcumm~ances the diecolouration occurs. Since the modification of the solenoid valve system [Sept. i0, 1993) dlscolouration of the menthol in the bath has not occurred. Monitorina of Humectant Levels in ~TL and ODDoeitic~ ~n~.: Glycerol levels were chocked for RJR-Macdonald and RBH products. bone dry-weight basis the glycerol levels for 1993 were: SAMPLE Craven "A" Mark Ten Zxpo~ A Expor~ A Light Export A Ultra Light Export A SPBL (Ii0 g) Expo~c A Light SYBL (110 g) Belvedere S,,peroll 200 (135 g) Belvedere SUP.200 (90 g) Belvedere SUP.200 Ex.M (90 g) Belvedere Eaey~ol Belvedere Ex.Mild Easyrol On a GLYCEROL (%) Zs~ 2nd 3rd 4th YEAR YEAR QTR. QTR. QTR. QTR. AVE. S.D. 2.66 2.87 2.91 2.77 2.80 0.10 2.63 2.57 3.02 2.61 2.71 0.18 3.34 3.87 4.25 4.02 3.87 0.33 4.89 3.90 3.6 3.12 3.88 0.65 3.72 3.87 4.98 4.54 4.28 0.51 2.67 2.49 3.22 2.35 2.68 0.33 I.i0 2.33 3.16 3.00 2.40 0.81 2.71 2.87 3.44 3.47 3.12 0.34 3.05 3.10 3.69 3.14 3.25 0.26 2.92 2.92 0.52 0.52 0.53 0.53 Six, sen samples of fine cut tobacco were tested for project T-4448 and T- 3219. To verify ~he quality of the samples before a subjective consumer test. To suppoz"c project T-3208 (D-59 tobacco) 8 samples from Corby were tested for glycerol levels. Glycerol levels were measured in samples before and after expansion to evaluate the glycerol losses during expansion. The average glycerol losses during expansion were: D-59: 14 ± 4% (n " 27)" • Average results for 27 determinations from Decembe= 1991 ~o August 1993. TO support the staz~-up of the new pilot plant, I0 samples we=e tested for glycerol levels. The average glycerol level was 3.9% ± 0.2% (C.V. - 4.39). The glycerol application was homogeneous and within specification. 5. Consumer ComPlaints Znvestiaations: In 1993, specific analyses were required for ten (10) consumer complaints (52% less ~han last year}. These complaints a=e listed in the table below: 0 u m B.A.T. CONFIDENTIAL - CATEGORY I: MINNESOTA TOBACCO LITIGATION BATCo document for PFSFC 1 March 1999

- 15 - Cowsm~R COMPLAI"W~S - ~.993 DESCRIPTION Cig~. flare up Gas s=ell Foreign matter Ho=rible taste Bad taste - irritation - Sick Oil in filter - Sick Foreign matter - insec~ ~tchy eyes and vomiting Strange taste Plastic taste and smell RESULTS SUBJECTIVE Nag. lqeg. Light unidentified off- Caste. Flat taste - Light cooling effect - Not sick. ANALYSIS May be a big piece of st"m. No Eoreign matter. Nag. Plant tissue Nag. Lacewing Corder Neuropatra|. Nag. Ntg. Nag. 7. Non-Routine Ex~loratorv Work for Product Develooment or ~or Gensr~1 T-'oubleshoo~ina in Plants= Zn July, analyses were performed to determine the composition of an oily brown (rusty) deposlt found in a filter in an air compressor llne in Aylmer Plant. Analysis showed that the dapoelt contained Essolube H.D. I0 W. oil, rust, plant tissues (probably tobacco), water and microorganisms (bacteria and fungi). A nl,4 dryer was installed in Aylmer Plant. In October, two production teats were performed. Two series of samples (Teat #I - 12 samples of H2S Grade tobacco and Test #2 m 25 samplee of AF2S Grade tobacco) were analyzed to detect oils and/or greases. No oils or greases ware detected in these tobacco samples° The limit of detection for the chromatographic method used is 0.01% W/W of oil in tobacco. Zn Oc~=ber the purity of new nicotine standards prepared on J-i for routine nicotine analysis was checked (98.6t). In October, a problem of build-up on Ohm knives of a filter maker in Montreal occurred during ~he production of 18-2 filters. The analyses of plasticizer levels in the filter rods revealed ~haU an excess of plaaticlzer was not ~he cause of the probl~n. The filter rods contained 2.85 g Pz/100 rods. The specification for 18-2 filters is 3.2 g Pz/100 rodS. O I B.A.T. CONFIDENTIAL - CATEGORY I: MINNESOTA TOBACCO LITIGATION BATCo document for PFSFC 1 March 1999

- 17 - RECORD TYPE: SUB T~.PE = SECURITY CODE: FUNDING BODY: ORGANIZATION = GROUP NUMBER= LOCAL PROJECT NUMBER(S)= PROJECT. TITLE: PERSON RESPONSIBLE: EFFORT: PROJECT DESCRIPTION: SCOPE: DEPTH: FUNCTZON: OB~'E CT'_VE: CLUSTER: DATE REVIEW WRITTEN= REVYZW TITLE: REVIEW TEXT: ITL CANADA T-7282 Reduc~cion of Irritation PORTF.R, A.; M¢RRZDE, C. 0.35 - 1993 A range of tobacco, paper and filter additives will be screened to assess thai=- sffe~ on reducing smoke irri~a~ion. The add£~ivee will also be evaluated for possible negative changes in smoke quality, s~abili~y, ¢os~ and ease of appLicat ion. GROUP RELEVANT FUNDAMENTAL GENERAL ALTERNATIVE PRODUCT RES/DEV. Janus=7 1994 Reduction of Irritation In :he last p=ogrese repor~ several approaches to i=T.i~ation reduction were described. The mos= promising approach for conventional products was ~o usa acid ~rea~ed paper. Some fil~er additives were effe~ive bu~ mos~ of ~hase weca no~ s~able for more ~han one to ~wo weeks, except Na~, which reduces irritation in DAY samples (with high formaldehyde deliveries} but increases irritation in conventional cigarettes. S~nce July, we have studied 3 absorbent falters: carbon, Duolite A-? and Diaion CR-20 resins. Carbon was used in a triple filter conflgura:ion an a loading of 75 mg. Duolite A-7, half neutralized with acetic acid, was supplied in a dual, dalmatian configuration (60 mq/tip) by B & W. These ware manufactured for S & W by Filtrona, U.K. in 1987 and the resin is no Iongsr ¢onsnercially available. Diaion CR-20 resin, in the free base form and half neutralized with acetic acid, or glutamic acid was incorporated by hand into cavity filters at levels of 50 and 1O0 mg/tip. The absorbent filters were a~ached to du Maurier K.S. cigarettes and smoked subjectively. All samples reduced Irritation wi~h carbon and duoline giving unacceptable off-tas:8. The age of ~he Duolite samples may have been a factor in the formation of off- tas~s since :his probllmwas no~ reported by ~ & W. Of :he differently :rested Diaion resin ~here was no off-taste with the free base while :he acid :rested samples caused a sligh~ change in smoke charac"cer. Aldehyde daliverles were de:ermined for du Maurier K.S. cigarettes with the free base and glu~aml¢ acid :reared Diaion OR-20 fil~ers: ~D Co B.A.T. CONFIDENTIAL - CATEGORY I: i~,IINNESOTA TOBACCO LITIGATION BATCo document for PFSFC 1 March 1999

- 18 - Calluloee ;mu:ocac= (conCz'ol) CR-20 f=te base ( 50 rag') CR-20 free base (100 rag) .. CR-20 + Glu~amic Acid ( 50 mg) CR-20 + Glutamic Acid (100 aw) Formaldehyde 97 23 I76) 19 (807 28 (71) 19 (80) J. Acmcaldehyda &¢z'olein (q :reduction ) 1122 555 (SZ) 282 (75) 698 (38) 386 (66) 134 80 (40) 49 (63) 91 (3z) 62 (54) ~zrChmc wock w£11 be ca~ied ouC ~o assess ~he effect of agtJlg on ~hese f£1cmc=. 0 P,,3 .4::,. Co "-,4 B.A.T. CONFIDENTIAL - CATEGORY I: MINNESOTA TOBACCO LITIGATION BATCo document for PFSFC 1 March 1999

- 19 - RECORD TYPE: SUB TYPE: SECURITY CODE: FUNDING BODY= ORGANZZATZON: GROUP NUMBER: LOCAL PROJECT NUMBER ( S } : pROJECT TITLE: PERSON RESPONSIBLE: EFFORT: PROJECT DESCRIPTION: SCOPE: DEPTH: FUNCTION: OBJECTIVE: CLUSTER: DATE REVIEW WRITTEN: REVIEW TITLE: ~TL CANADA 416 T-9642 Development of a Procedure for ~he Determination of Possible Contaminants in ToBacco and Cigarette Samples POULZN, P.; DUMONT, J. 1993 To develop as ex~ensive as pQs|lhle analytical procedure for the analysis of allegedly contaminat~ samples. This will ensure a better avalua=ion of consumer complaints and will assist ManufaCturing when an investigation of an oil or grease leak is Ee~ui=ed. LOCAL SUPPORT GENERAL SUBJECTIVE METHOD DEV. January 1994 Development of a procedure for ~he determination of possible contaminants in tobacco and cigaret~8 samples. REVZEW TEXT= The existing da~a ~ale (chemical and physical characteristics, ZR and OV specU~a and chromatogra~s) of the oils, greases and solvents used in the plants are being upgraded with GC/MS scans of the volatill f=a¢~ione. This wo=k is an ongoing project. ¢D rx~ ~D cc CC B.A.T. CONFIDENTIAL - CATEGORY I: i'~IrNNESOTA TOBACCO LITIGATION BATCo document for PFSFC 1 March 1999

- 20 - RECORD TYPE: SUB TYPE- SECURITY CODE: FUNDING BODY: ORGAN~ ZATION: GROUP NUMBER: PROJECT NUMBER (S) : PROJECT T'r'~: PERSON RESPONSIBLE: EFFORT: PROJECT DESCRIPTION: SCOPE: DEPTH: FUMCT$ON: OBJECTIVE: CLUSTER: DATy- REVIEW WRITTEN: REVIEW TITLE: ITL CANADA 416 T-5549 Review and Development of Al~arnatlvt Analy~iual Methods POULIN, P.; DUMONT, J. '1993 The original objective was ~o develop chroma~ographl= methods rala~ed ~o the nature of ~he compound to be analyzed (GC for volatile c~mponenus, HPLC foe non-volatile). The objec=ive has been axpandld to include &naly~ical methods in general ~o meet the changing requirements of R & D. LOCAL SUPPORT GENERAL SUBJECTIVE METHOD DEV. January 1994 Revi~ and development of alternative analytical mathoda. REVIEW TZXT: i. Evaluation of a ~ew P1asticlzer AnalyzeE A new plasticlzer analyzer, the FPZ-1O0 f=c~ F~DUS (Filtrate) was tvalu&tmd for a peeled of two ~onILheo The FPZ-100 is built uo be used in a p~ant area. I~ uses NIR (Hear Infrared Reflectance) to measure a sample's diffused reflectance. This instrument represents an improvemsn= in plasUlcize= analysis. The advantages are: - No solvent - Non-destruc~ive method - Instrument easy to use - Results obtained in 60 seconds - NO weighing o= sample preparation - No dry rods to prepare - No inue~Eupuion of the KDF ~o prepare dL'~ rods. The disadvantages a~e: - The prnparation of standard filter rods is long and ~aborious - The lifetim~ of a callbEation curve is uncertain - The wet and dry methods p~esently used in the plan=s give comparable results - A qualified lab ~ech~i¢ian ha8 UO calibrate the instrument - Sample presentation area and the keyboard should be improved. From ~he results of this evaluation, at leas~ two calibration tu~vee would ~e needed to cover all filter rod production. This represents a day of work in the plant for preparation of ~he standard filter =ods and ~he requisition of a KDF, 6 days of lab work to determine their reference values and another day to calibrate the FPZ ZOO and make lONe verifications. According to the |ales representative of FIDUS, this process has ~o be repeated every $ months. This rep=esen~s approxima=sly 16 days/person~year foe Non-Routlne AnalT~ical Service. Maximum curren~ 8ffor~ is approximately 4 days/pe~lon/yeaE a~d no i~creasa in lab effor~ in envisioned in ~he foreseeable future. CD ~C C~ ~D B.A.T. CONFIDENTIAL - CATEGORY h ~IlNNESOTA TOBACCO LITIGATION BATCo document for PFSFC 1 March 1999

- 21 - Therefore, el=hough =ha FPZI00 ks accurate (within • 5 m~/rod) and reproducible, once calibrated, the effo~'c required to sen up the instrument for in-plan= use limits its usefulness. 2. Annual Audit of Anal~*cical Needs and Methods qqed ~ R & ~: A new ~eth~d using GC/MS was developed =o determine the purity of l- menthol c~stals. The iJ~puri~ies found in chinese menthol (Polar Bear bra~ds) we e, = - (-)-Zsopulagol, p-Menthone and nee-Menthol. ~m~roved Me, hod lot 8enzo(a1~vrene: A method was developed at ITL in 1989 (ITL Method #28) to analyze B(a)P in WTPM. This procedure allows the determination of approximately 2 sampZes/day. This severely limits the feasibility of using this method foe routine analysis. In method #28, the =at is collected on a Cambridge pad and B(a)P is extracted by shaking in cyclohexane (I mL/l ~g WTPM), filtered and evaporated =o • i0 mL. This filtrate is diluted to i0 mL with cyclohaxans, sonicated and re-filtered. The sample is =ban ready to be cleaned c~er an NH: Sep-Pak cartridge. After the Sep-Pak has been conditioned with hsxane, an aliquot, of I-4 mr- of the filtrate is added to the cartridge. The method has been streamlined further using 40 mL of =y=lohexane regardless of weigh~ of WTPM. With this ~odlflcatio~ It is possible t= avoid the evaporation step. After the Sep-Pak has been conditioned, an aliquot of 4-16 mL of the f£1trate is added =o the ca:~rid~e. The results obtained by the two procedures are as follows: Method No. of Trials- B(a)P in FTC Tar ( ng'Im~ ) 4 0.7 ± 0.1 8 0.83 : 0.06 I =L cyclohexane/1 WTPM 40mL cyclohexane -Five IR1 ci;a:ettes were smoked per pad. BIa)P Delivery fnglci~) 26 ± 4 28=2 According to a student's t-test (p • 0.05), =hera are no statistical differences (p • 0.05) ~unong the results wi~h either procedure. This method can be used to determine 4-6 pads/day for benzo(a)pyrene analyses, which allows the method to be used for rouclne analyses. These are preliminary relults and must be confirmed. ~mvelo~ent of a ~e=hcd foc the Analysis of Free Amlno Acids ~n Tobacco: We have =eceived a new reagent for the analysis of amino acids recently (Nov. '93) c~erclalized by Waters. The reagent is 6-Aminoqu£noly1-N- hydroxysuccl.uimidyl carbamate (AQC), and works with primary and secondary amino acids. The reagent will be evaluated in the new year in teems of accuracy, precision an~ suitability for ~obacco analysis, and the resu~a will be compared wi=h other procedures. CD B.A.T. CONFIDENTIAL - CATEGORY I: MINNESOTA TOBACCO LITIGATION BATCo document for PFSFC 1 March 1999

- 22 - RECORD TYPE: SUB TYPE : SECURITY CODE: FUNDING BODY: ORGRJgI ZATZON: NUMBER= LOCAL PROJECT NUMBER ( S ) : PROJECT TITLE: PERSON RESPONSIBLE: EFFORT: PROJECT DESCRIPTION: SCOPE : DEPTH: FUNCTION: OBJECTIVE: CLUSTER: DATE REVIEW WRITTZN: REVIEW TITLE: ITL CANADA T-S678 Developa~Bnt of a P--one=el I::o Establish Residual 5olvenu Upper Limius in Packaging Manorial. DUMONT, J.; DUNN, R. ; LETOURNEAU, C. 1%93 Development of a protocol to establish residual solvent upper limits in packaging material. GROUP FUNDAMENTAL GENERAL January 1994 Developmenn of a protocol to establish residual solvent upper limits in packaging material. REVIEW TEXT: A protocol has been developed to establish upper limits for residual solvents or compounds in packaging maUsrial. We have also investigated how Chase solvents or compounds affect the Caste of tobacco products. An analytical p:octdure was developed to detemine residual solvents. Bun it was realized ~hat monitoring only solvent levels was nor sufficient to fully ¢haracnerize packaging materials with an unusual odour. The printing process may include nu~ny steps, much as ¢lay coating, laoquer, ink applications, ovsrlacquer, etc. Each step ham the potential for odour devmlopa~Inn. It is non clear yet if all these steps contribute to the taste modification of tobacco produc~s, but in the development of a protocol for residual odour, it was thought necessary to monitor all volatilea ~han may be eminned from packaging material. The protocol for subjec=iva evaluation has also been evaluated. Two dlffersnn exposure modes {sandwich and cylinder) warm compared. BoUh were Csscsd in terms of solvent transfers to cigarettes. The mxposurs oondlCions Ctmml~erature, duration} were also avaluaUed. Both modes are equivalenC in terms of solvent transfer. However, the second mode is preferred because it is more representative of a pack of cigarettes, where the cigarettes are separated from packaging material by aluminum foil (no direo~ contact). Evaluation of the upper limits for individual residual solvents has been completed. Solvent comJ~ination8 either have no effect on or lower the individual upper lln~ts depending on the combination of solvenus. The results arm presented in Table Io A report will he issued in February I%94. O m ~D B.A.T. CONFIDENTIAL - CATEGORY I: MINNESOTA TOBACCO LITIGATION BATCo document for PFSFC 1 March 1999

- 23 - SOLVENT NAMZ Zr.hanol Zsop~-opanol ! Et:hyL Acetate lsopL-o~y 3. Acer.aCe P~pyl Acetate " subjective Evaluation Resul~9 I MZXTURZ MXXTURE 1 2 457 931 51 -- ) sg 119 38 (a) (b) t t - (a)= (b): (c): I MIXTURZ 3 465 60 (c) INDMDUAL ~PPER LZMITS 91S 102 20 117 76 NoCioea~ly mOdifi~ Baste with slight off-Caste. Off-Caste and modlfla~. No off-Baste. Sligh~ly flatter and m~ighCly pep-pezT° CD B.A.T. CONFIDENTIAL - CATEGORY I: MINNESOTA TOBACCO LITIGATION BATCo document for PFSFC 1 March 1999

- 24 - k'ORX ~ O3 SHOF,.ZNG 3ZH.x.VZ OUR 0 r~ ,,.,.a. B.A.T. CONFIDENTIAL - CATEGORY I: ~IINNESOTA TOBACCO LITIGATION BATCo document for PFSFC 1 March 1999

- 25 - RECORD TYPE: SUB TYPE: SECURITY CODE: FUNDING BODY: ORGANIZATION: GROUP NUMBER: LOCAL PROJECT NUMBER(S): PROJECT TITLE : PERSON RESPONSIBLE: EFFORT: PROJECT DESCRIPTION: SCOPE: UE?TH: FUNCTION: OBJECTIVE: CLUSTER: DATE REVIEW WRITTZN: REVIEW TITLE= REVZEW TEXT: ITL CANADA 441 T-8079-I;8079-2;8077-17;8077-18;8077-20; 8080-1;8080-3 Smoking Behaviour YOUSSEF, M. 2 " 1993 The objective of the smoking behavicur progr~,- is tO provide a scientific understanding of the interaction between the smoker and she produ~. To this end the recording system is being upgraded for improved ease of use, precision and rate at which data can be generated (T-8079-1). Changes in smoking behaviour resulting from product modifications will be inveetiganed (T-8080-1). GROUP RELEVANT FUNDAMENTAL MARKETING; PUBLIC AFFAIRS SUBJECTIVE; LEVELS SMOKING BEHAVIOUR January 1994 Smoking Behaviour ~oduc: Elasticity It ham been suggested t~t one way of increasing produ~ elasticity ac a given pressure ~op el to increase filter pressure drop and rtducm tobacco rod prRssure d~'op. We are invesnlga~ing this hypothesis by mmaBuring smoke ~liverie8 at different puff volumes for specially designed samples. La=er, if ~ results suggest differences Ln elasticity, a smoking behavi=ur analysis will be initiated. A series of cigarettes was designed with different tobacco and filter rods. Tobacco ~de were made to du Maurier Ultra Light K.S. (duMLTLXS) specifications at densities ranging from 20-27 cg/cc in order to have a broad range in rod pressure drops. The rods, eele~ed for pressure drop, were smoked to determine deliveries. Filter parameters {pressure drop, ventilation) were then selected in order to make cigarentes with equal standard deliveries but with different rod and filter pressure drops. The cigarettes were smoked at puff volumes of 3S, 50 and 70 ml to determine elasticity. If the tar delivery increases in direct proportion to ~he increase in puff volume, the produc~ is inelasnic (i.e. elasticlty = I), while if tar delivery increases faster than puff volume, elasticity > I. Two other cigarette samples were in the study: du Maurier Ul~a Light K.S. as a control, and Merit (Philip Morris), which is believed to be elastic. Results ere denailed in Table I. None of the samples except for Merit were elastic. This is probably due to the similar levels of ventilation between the experimental samples, and the fact that there is not much variation in the filter and rod pressure drops between the designs. A second potential approach to achieving elasticity is ~o use the Gap filter (FXL Znternational, U.K.). Two series of samples were made with Gap filters: One series wan made to duMULKS sp~clfications with a control filter (porous plugwrap, pre-perforated tipping), and two types of Gap filters, both with 7 gaps but with T-settles p.d.s cf 88 mm and llS mm respectively. The other series were made to Matin6e Extra Mild KS specifications using the same filters as above. CD %C B.A.T. CONFIDENTIAL - CATEGORY h MINNESOTA TOBACCO LITIGATION BATCo document for PFSFC 1 March 1999

- 26 - The Gap clgaro~tes weEe peEforsted manually, using a heated pin, to match the level of ventilation of duMULKS and MatXMKS control ¢igarettes. FOE each ¢igauret~e sample, ventilation vs flc~ Ease and delivery ve puff volume were studied. Results are detailed in Tables II and IIZ. For all samples ventilation de==eased linearly with incEeaeed flow =sUes bu~ ~rQ Eapldly for Gap ¢igarettee compaEed to control cigarettes. Within each series u~e control and Gap cigarettes have similu deliveries at 35 ml puff volume. The Gap cigarettes were more slavic than the contEol8 at 70 ml puff volume. Ad ~oc sub~ecl:ive comparison of cigaz~ttee #244 and #436 showed a definite incrsase in smoke s~=angth with cigaEe~=e.#436. Further work will include the examination of on-line laser perforated gap clgareCtee and a smoking behaviour study of these cigarettes. TABLE X Sample I duKOL 520 904 622 695 Merit Fil~er PD (=m) 79 84 8"7 79 73 151 Rod PD (a) Ciq~:. DR {m) I Vent ' n (t) 35 Tar 7.7 7.7 7.5 8.8 8.2 1.7 68 45 54 55 97 50 Deliveries at =~ puff vo~ Nic. PN 0.9 10.5 O.S 8.7 0.9 9.3 0.9 i0.I 1.1 11.5 0.2 6.4 96 i00 104 96 I00 i09 52 42 45 46 51 61 TR~LLE IZ Elasticity (~ased on TeE a~ 70 ml) 1.05 1.03 i.I 1.03 1.05 1.30 U Flow Applied (eel=in) 650 850 1050 (Std.Coad) 1250 1450 1580 co::esp,:md~ Volume (¢=) 21.6 28.4 35.0 41.6 48.4 52.6 Ventilation (%) duU4ULKS MatXMKS Con=tel Gap Cir. Low P.D. 58 55 53 52 51 63 56 52 50 47 Gap Cig~c. High P.D. 58 54 50 47 44 60 60 59 57 56 55 Gap Cigt. Low P.D. 67 63 59 56 53 52 Gap cage. High P.0. 66 61 58 55 51 49 CD w Lm ,.o Lm B.A.T. CONFIDENTIAL - CATEGORY h NII'N'NESOTA TOBACCO LITIGATION BATCo document for PFSFC 1 March 1999

- 27 - saeple Con~z'o i (244} Gap cirri:. Low P.D. {436) Gap cig"=. High I,.:). (120) 35 70 35 70 35 D.R. Ven~'n Ta~ Nic. Puff Tax" (mm H:0) (t) (2) !mg) So. Elasticity - $0 7.2 0.7 10.4 93-10S 51 14.6 1.4 9.3 1.01 66-76 59 7.5 0.8 10.5 65-73 59 16.9 1.5 9.3 1.13 51 8.2 0.7 10.6 Ca3nurol 35 87-97 55 S.l 0.S 8.7 (942) Oap Ci~. 3S 79-89 S4 6.0 0.6 0.51 LOw P.O. (62S} Gap Cig~. 35 88-66 54 6.0 O.S 8.6 High P.O. 70 04"95 54 13-1 i.i 7.5 {2061 i.ii CD On %O O~ B.A.T. CONFIDENTIAL - CATEGORY I: MINNESOTA TOBACCO LITIGATION BATCo document for PFSFC 1 March 1999

- 28 - WORX ~ 04 BIQr.~:~ 2 CA,~, RZ 5 .E,~,.RCH B.A.T. CONFIDENTIAL - CATEGORY I: MINNESOTA TOBACCO LITIGATION BATCo document for PFSFC 1 March 1999

- 29 - RECORD TYPE: SUB TYPE: SECORZTT CODE: FUNDING BODY: ORGANIZATION: GROUP SOMBER: LOCAL PROJECT. NUMBER ( $ ) : PROJECT TITLE: PERSON RESPONSIBLE: EFFORT: PROJECT DESCRIPTION: SCOPE: DEPTH: FUNCTI ON: OBJECT IVE: CLUSTER: DATE REVIEW WRITTEN: REVIEW TITLE: ITL CANADA 445 T-7708;04 BioLogical Effects of Tobacco Smoke and Tobacco Extracts in Short TeEm Tests BIL~HORIA,M.H. 1993 Bacterial teats will be employed to study the mutagenicity of smoke condeneatee fr~ ITL and opposition brands, as well as new market entries, to ensure that ITL products rank favouEably in a comparative study. By determining the mutagenicity of smoke condeneates and fractions from different cigarettes smoked under different conditions, the aim is to identify thosm parameters which affec~ mutagenicity. The effect of additives on condensate mutagenicity will also be studied as will smokeless tobacco products. GROUP APPLiED/DEVeLOPMENT PUBLIC AFFAIRS REGULATORY BICLOGY January 1994 Biological Effects of Tobacco Smoke and Tobacco Extracts in Short TeEm Teats. REV~EWTEXT: i) Protein content and oxidation state o~ tobacco, and the mu~auenicitv of ciqarette smoke condensatw: In the previous semi-annual repor~ we showed that a single sample of groin, uncured tobacco showed a very low specific mutagenic activity (specific activity s 0.41} even though its protein content was comparable to a flue- cured sample whose smoke gave a specific Ames activity of 1.89. However, this evidence was weak on account of the different oE£gina of the cured and uncured samples, we have now repeated this study using uncured tobaccos from dlffeEent plm',~c positions from a single farm which also provided the corresponding cured tobaccos. The fresh, uncurtd tobaccos were d:ied in an oven overnight at 50"C. The cured and uncu:ed tobaccos were cut and cigarettes made by hand. These were tested for Ames mutagenicity by the standard procedures employed £n ~he laboratory. The p~tein content of the tobaccos was determined by a recently developed direc~ apectrophotometEic procedure. 0 m (..n co B.A.T. CONFIDENTIAL - CATEGORY I: I%,IINNESOTA TOBACCO LITIGATION BATCo document for PFSFC 1 March 1999

- 30 - p~oT'ZZ~ co~:"z~rT or 1~mAcco ~ ;qro~Olp~ZCZTY o1,' sxox~ .com)~:~sx~t TOBACCO LUG GRADE Uncured Flue-Cured C~.~ GRADE Uncured Flue-cured TIP GRADE Uncured Tlue-cured PROTEIN NITROGEN • DWB 0.89 0.81 1.37 0.80 PROTEIN CONTENT • 9W~ 5.23 4.21 8.83 4.62 1.87 1.08 14.85 6.00 AMES MUTAGENICITY SPECIFTC ACTIVZTY 0.9S 2.!I 1.57 1.93 2.18 2.85 IgG as s~andard Fr~ ~he results og this study presented Ln Table 9 it will be seen ~ha~ tn every =ass the cured sample was much lower in protein ooncen~ bur si~ifi=antly higher in muuagenio activity. For example, ~he uncured lug grade oontained 5.3% protein while the cured lug grade contained 4.2% protein. However, when mutagenici~ios are compared for the lug grade, the cured sample at 2.11 mpeoific a~ivity was much higher than =he uncured sample which recorded a 0peciflc mutagsnic ac~iv£ty of 0.95. This is illuau=ated more clearly when 8~¢ifi= activity is expressed per ~ pEG=sin. Such values for &'=.Ted tobaccos range between 0.42 and O.S0, and are aJ~ou~ these time8 higher tha~ similar values for uncured uobaecos where they range be=wean 0.14 and 0.18. Thus, this study confir=~ our earlier observation the= the oxida=lon ~a=e of =obacco too, plays a role in de~e~mining ~he mu~agenicity of omoke condensers. It will be interesting ~o date=mine if other tobacco varieties which are cured differen=ly give similar resul~e. 2. A s~u~ ~¢ ~ew "Elite" Tobacco Strains Produced by Aorlculturs Canada - Prc~e~ Con~%nt of ToDacco add ~u~aq~qicitv of Smoke Condensate: The Delhi Research S~a~ion of Agzi~ul~re Canada ham an on-going progranc~e of evolving "eli~m" tobacco s~caine =harac=erized by ~heir adaptation ~o Ontario growth conditions as well as by nhelr improved yield, leaf quality and organolop~io prope~ies. Some o~ theme tobacco s=raina have been sent co ITL for ¢igarsc=e manufa~ure and distribution for organolep~ic tea=ins. We have tested these cigarettes fo~ Ames mutagenicity and protein content. The results of an examine=ion of some of the more promising tobacco s~r~ine ~ro~n~ in 1991 are presented in Table 10, while those grown in 1992 are presenued in Table 11. ~D ~D B.A.T. CONFIDENTIAL - CATEGORY I: MINNESOTA TOBACCO LITIGATION BATCo document for PFSFC 1 March 1999

- 31 - PROTEIN CONTENT OF &ORICtTLT~?~ CR~AD&"wr'rTE. " TOBACCO (199~) STRAINS'¢ ~rD ~JT~GE~YCZTY OF SMOI~ CONDENS~'~ TOBACCO PROTEIN CONTENT - mcl/(:,~r t DWBI" PLCX 28 873 360 (Delhi 76) 177 (Delgold] 474 530 ~IgG as 8=anda~d. MUTAGENICZTY OF CONDENSATE - SPECIFIC ACTIVITY- 44.17 : 1.4 44.17 : 1.4 43.50 = 1.3 42.50 : 0.0 1.00 ± 0.23 1.91 ± 0.47 1.59 ± 0.43 1.43 ± 0.35 T~ Z~ PROTZTX CONTENT OF AORIC~LT~I~E CANADA "ELITE" ~OBACCO (1992) STRAINS2 AND MUT&GENICITY OF S~OK~ CQ~DENsAT~, TOBACCO BRR INDEX 8.5 8.2 4.3 4.9 5.0 151 420 534 S60 658 Delgold Control PLCK 28 t IgG as sCandard. PROTEIN CONTENT OF TORACCOt - ~/~ nws 43.50 : 1.3 46.00 : 3.0 44.33 : 1.2 45.83 ± 1.4 45.83 : 1.4 44.17 : 1.4 MUTAGENICITY OF CONDENSATE - SPECIFIC ACTIVITY 1.87 -~ 0.14" 1.61 -~ 0.16' I. 55 ± 0.11b 1.49 ± 0.12b 1.37 ± 0.19" 1.00 ± 0.06' • Mutagonic activities followed by different superscripts are signLficantly diffe=enU (p < 0.01). BBR - Black Root Rot CD CD CD B.A.T. CONFIDENTIAL - CATEGORY I: MINNESOTA TOBACCO LITIGATION BATCo document for PFSFC 1 March 1999

- 32 - It w£11 be soon that all ~he 1991 new ~obacco s~rains as well ae the controls (Delgold and Delhi 76) were similar An terms of tobacco protein and smoke mutagenlcity, wluh Delgold being the lowest and 8igniflcantly different from strain 873 In fezes of Ames mutagenlcity. $imilarly, gelgold was e£ffnlfican~ly lowe~ An Ames mutageniclty than all the 1992 samples. More Interesting ks the comparison of mutagenlolty he,won Check 28 and the n~ elite redact, ¢igare~es in which the lat~er are 40-90, highe~ than the Chock ¢ig~tte. Inclusion of 19% stem and its plaln-end ten,fuSion wou2d pax~lally aoeounu for such a difference. However, other fa~ors 8u¢~ am different revels of Ee~ox eompound~ may account for some of this difference. It should be pointed ou~ ~ha~ Agriculture Canada strains grown in 1992 were also tu~ for thei~ resistance .to black root ro~ disease and 8ignlficant differences noted. 0 m r~ c B.A.T. CONFIDENTIAL - CATEGORY I: ~IrNNESOTA TOBACCO LITIGATION BATCo document for PFSFC 1 March 1999

- 33 - RECORD TYPE: SUB TYPE: SECURITY CODE: FUNDING EODY- ORGAN$ ZATION- GROUP NUMBER= LOCAL PROJECT NUMBER ( S ) : PROJECT TITLE : PERSON RESPONSIBLE= EFFORT: PROJECT DESCRIPTION: SCOPE: DEPT~: FUNCTION: OBJECTIVE: CLUSTE~: DATE REVIZW WRITTEN: REVTEW TITLE: REVIEW TEXT: ~TL CANADA 445 T-6775;04 Microbiological Examination of Canadian Tobac=o Products, Tobacco Processing and Raw Materials B ILIMORZA, M. H. 0.3* 1993 Standazd microbiological procedu:es for enumerating a variety of microorganisms will be employed to control the microbiological ~uality of ITL produ¢~:8. Areas of concern rage=cling hygiene iA t~e plant will also be investigated. LOCAL SUPPORT GENERAL REGULATORY BIOLOGY Januazy 1994 Microbiological Examination of Canadian Tobacco Products, Tobacco Processing and Raw Materials. (I) Process temperature chance dur~nc flue- cuz~Mist=~= condItionina and its effec~ o~ the microbio~OqV of the strips= Timm-tlmpeu:ature relationships in teems of effect on the microflora of tobacco strips during p~oceeeing have been studied in both the Montreal and Guelph plants. Since ~here is no aiz-ccnditioning in the Guelph plant, this study has been cazried ou~ in the winter as well ae summer seasons, since high ambient temperature and relative humidity a~e bound to play a role in the keeping quality of tobacco s~rips during processing. Temperatures in Guelph in r.he summer mon~hs aze known to exceed 90"F during the day. The renul~s of ~he winter s~udy have been presenu~ in the lasB progress report. The results of the summer s~udy are presented in ~his rspor~. From ~he Montreal results p=asented in Table I, At will be seen tha~ ~hs mlcEohial pic~:ure appears normal and similar ~o ~ha: recorded in the earlier s~udiee. In fac~, there appears to he a decline An the mould flora au the higher processing temperature. The time for which this batch was held at the higher temperature before transferring ~o the blending bin might explain ~he low mould counts. The results of the Guelph study ~ze presented in TaBle 2. Once aga£n, i~ will be seen Bhat there was no~hlng remarkable about ~he microbiology of the tobacco £n both the lower as well as higher temperature studies. Finally, it should be poln~ed out ~ha~ ~he studies aE bo~h Montreal and Guelph could only he undertaken in the month of June, whicB is not che waemesB month of the ye~0:. While high temperature trea~4~en~ of tobacco at the CTCM/CCM IlagI might riduce the micrcflora, increases in microbial counts might result in the blinding bins during periods of high temperature and high humidity during production in the hottest months of the yea=. 2. pct~$1um Sorbate Concentra=£on an~ Shelf-life of Orville Pine Cut This is par~ of a continuing study to determine the effec~ of sorbate concentration on mould development on fine cut tobacco. CD ~o ~n P~ ~o B.A.T. CONFIDENTIAL - CATEGOR.Y I: ~[INNESOTA TOBACCO LITIGATION BATCo document for PFSFC 1 March 1999

- 34 - Fine cut ~.obacco was s'r.orIKl under amb~en~ oondi=ions: 1) ~ =~e =eeeazch laJ3o~&toL'y and 2) in the pile= plant. The fine cur stored in ~he lahorato=~' was examine~, up ~'.o g months, a.nd. no moult/ was detsc~'.ed e£~.he= in : visua~ examination o: by smmlling :he produc~. The o=he= batch of fine cu~ tobacco scored in ~he pilo¢ plAn~ (50 boxes of fine cur tobacco a~ etch ].evtL of sorb:~'.s) was examtne4 &f".e: 16 ~nths. Zn ".he cont'.:ol. fine cu~ tobacco (0.073t po:aceium sorba:e), 9 boxes cu= of 50 showed mould growr..h and an a~noEmal smell. At 0.182% sorbal=e all SO boxes examined had a normal s~-ll ~4 no v£sLble mould. A~ ~h~ ~vo ~igher concen~:ations of so:bate (0".333t and 0.532t) only 20 boxes seth ou: of ~e 50 boxem, were exaa~ned tad found to be free of mould and bad odou:. The results of ~his study |u99em: the= £f ~he level of F~assiu~ sorba~e was inc~ased to at leas1= 0.18, in all cue fine cuts, mould growth would be :s'~uded for over a yea= under ambient conditions. Presently. potassium so,bate is added at ~he level of 0.21, before the ~cbacco is DIET-pEocesse~, after which the level of eorbate drops ~o @ouc 0.07q. Adjusting the level of ~oEba~e to obtain ah~u~ 0. ISt in the final produc~ w~uld improve i~s keeping c~ali~y. CD B.A.T. CONFIDENT[AL- CATEGORY h ~IINNESOTA TOBACCO LITIGATION BATCo document for PFSFC 1 March 1999

O ~O "n "n "I O _x ~o O Z o m z I O P-4 e, m L~ o o co > N 0 0 ,-'t ~4 o z - )5- ~;r TEHPERA~RB ON TIIB H|CROfllOIJDQT Or FLUE-CURBD ~). BACCO BTRIP8 C ONDIT|ONE;D AT 20t HO|8~rJRI~ FOR i DAYS IN HONTRSAL PLANT IIOLDING TIH£ IN BLENDING BIN (D~YS) RgCZPE C-09 0 1 2 3 4 ~ECIPE C-2~ O 1 2 3 4 TEMPERATURE (HOISTURE| OF TOBACCO STRIPS 30.5"C (20.St) 26"C (19.76) 27.5*C (19.6t) 27"C ()9.31) 26.5°C (19.?t) TOTkL 6&CTERIR BRAIN HEART INFUSION AGAR 2.76 x IOs 3,92 x 106 1.98 x 10l 5.26 x 10s 9.47 x 10j COL I FOR/IS HoCONKEY BROTii < 1 Nil Nil Nil < I THgRHOPHILIC ACTINOH¥CETES NOVOBIOCIH AOAR 236 256 328 297 249 YEASTS LITTHAN OXGALL AGAR 94 643 5B 50 183 38"C 121.7t) 36.5eC (20.3t) 36° {20.3t) 35.5"C (20.4t) 3S'C (19.It| 3.25 x 104 6.04 x 10j 8.16 x l0s 2.05 x 106 1.46 x 10i Nil Nil till Nil till 411 < 25 461 17 566 13 289 < 25 411 < 25 The microbial numbers given are per gram tobacco. * Tobacco extra CTHC at 62.5°CI Tobacco exito Coolar at 26e-31.SeC. I Tobacco £xita C~HC at ?1.5°C; Tobacco exLto Cooler at 40e-42~C, HOULDS LITTHAN HALT OXGALL SALT AGAR AGAR 283 17B 166 294 100 89 266 255 147 78 < 25 17 33 25 < 25 225 17 17 56 < 25 .~ 1;,0 ?. cj 11;,Z 01;,

m oo O :T ~O O -n p.q > r" I > o w-4 o > O > CI 0 ,-4 > o z - 36 - KFI'EC~ OF TEMPERATURE ONTUB MICROBIOI,OOY OF i~UB-CURBD TOBACCO STR|p~ CONDIT~OHED AT 2Q~ MOTSTURE FOR 4 DAYS IH OU~PB PLANT HOLDZNG TIHE IN BLENDING BIN (DRYS) ~EC!PE c-24 RECIPE c-24 TEMPERATURE (MOISTURE) OF TOBACCO STRIPS 40.1"C (20.6q) 33,5°C (20.01) 34,3°C (19.4%) 34.70c (19.3t) 34.4"C (19.4t) BRAIN HEART INFUSION RGAR 2.12 x 10s 5.43 x 10j 5.31 x l0s 1.51 x 10s 7.89 x 105 COLIFORHS McCONKEY BROTH < 1 Nil < 1 NIl Nil TMRRHOPIII LIC ACTI NOHYCBTES HOVOBIOCIN ROAR 183 211 155 355 283 42.2"C (20.9t) 43.60C (19.9t) 41.8"C (19.6t| 41.50C (19.6%) 44.9°C (19.1t) 5.29 x 104 7.95 x 10j 1.11 x 106 9.14 x IOs 6.39 x 10' < 1 177 Nil 272 < 1 322 HLI 433 Nil 372 The microbial numbers given are per gcam tobacco. Tobacco oxLto CTHC at 72.8'C| I Tobacco Exits CTMC at 74.8"C; Tobacco exits Pro-Blonder at 38.4"C. Tobacco exits Pro-Blender at 42.5"C. OXGkLL RGAR I,416 850 250 743 2,761 556 233 893 78 25 MOULDS LITTHAH MFJ~T OXGALL SALT AGAR AGAR 200 139 300 128 139 ~ 122 183 72 183 183 J, 133 78 33 56 61 Nil 244 255 589 516 I t t77017

- 37 - O B.A.T. CONFIDENTIAL - CATEGORY I: MINNESOTA TOBACCO LITIGATION BATCo document for PFSFC 1 March 1999

- 38 - RECORD TYPE : SUB TYPE: SECURITY CODE: FUNDING BODY: ORGANIZATION: GROUP NUMBER: LOCAL PROJECT NUMBER(S); PROJECT TITLE: PERSON RESPONSIBLE : EI~FORT: PROJECT DESCRZPTZON: SCOPE : DEPTH: FUNCTION: OBJECTIVE : CLUSTER: DATE REVIEW WRITTZN: REVIEW TITLE: ITLCANADA 2429 T-0581 Creation Of Breeding Populations fr~ Somatic Hybrids. DUPLESSZS,G./BAI, D. 1993 The objective of the week iB to ¢rea~e breeding lines through recurrent backcrossing and sele~ion of somatic hy~Eide w£th ¢~rcialN. u__abacumcul~ivars. This work is in collaboration with the Delhi Research Station of Agriculture Canada. GROUP FUNDAMENTAL:APPLZED/DEVELOPMENT LEAF AGROTECHNOLOGy LEAF January 1994 C=sauion of 3r~ing Populations from Sciatic Hybrids. REVIEW TEXT: I. Radiation Treatment of the debneT~ Chromosome Addinion Line9 £n Tobacco w£th Resistance ~o Blac~ Roo~ Ro~: Plan=s wi~h resistance to blaGk root rot ~Thielav£o~sis basicola) (infection type, immune) we~ sele~ from the advanced backcross progeny of the s~atic hybrids between Niconiana ~abacum (4X = SSTT = 48) end N. debnevi (4X - ZZTY - 48). The selected plants all carried an extra pair of ~ chromosomes and it wee assumed nha~ the gets(s) for resistance no black root rot were on ~he debnevi ¢~omoeomee. Alnhoug~dn_dnebnevi chromosome addition lines wi~h resistance no black Eoo~ ro~ have been produced, the added debnevi chromosomes were transmi=~ed to ~he offspring an low frequency wi~h only 25% self- pollinated progeny of an addition iane carrying the extra ch=omosomes. In older to s~abili=e the resistance, Uranslocatlons between ~he added~ ohromosome frogmen= carrying the resistance gene(s) and ~he corresponding Eaglet of a ~obacoo chromosome have to be induced. A radiation treatment ~ae designed to achieve this goal. The seeds f:o= ~he addition lines were treated at AECL (Chalk River). As the gamma ray dosage increases a greater probability of obtaining a ~ranelocation is expected; however, the germination rate of the ~reated seeds decreases with dosage. The surviving seedlings were ~SS~ed for black root Lot resistance in the standard ~ank test and resistant plants were selected for selling. Screening foe tEanalocanions will Be ca=tied out in ~he M~. populations. 2. ~.~pD Markers for ~he Charac~q~iza=ion o~ debnev£ Chromosomes Carrying Black Raon Rot Resistance in Tobacco Add£~io~ ~nee: More Eel/ned methods t~an cytology are necessary no characterize Che debnevL chromosomes An the black Eoo~ rot resistant addition lines in Order to effloien~ly idennify ~ranslocanions involving ~ and ~obaoco chromosomes. Four hundred ~ando~ primers were investigated by generating RAPD markers using the polymerase chain reaction. Six~een random primers generated unique DNA fragments ranging from ~50 to 1600 bp (RAPU ~rkerS) and those markers were a~ribu~ed ~o the added ~ chromosomes. These RAPD markers will he used in monitoring transfer of the ~ ¢hE:,:OSome fragmen~ carrying ~he resistance gene into tobacco genomss. T~e same CD B.A.T. CONF:DE~T~AL - CATEGOP.Y h ~[INNESOTA TOBACCO LITIGATION BATCo document for PFSFC 1 March 1999

- 39 - =echnology can ~hen be used ~n in==ogression experimenus involving soma=it hybrids from other species. 3. Tooting of Disease Resistance Genes using RAPD Markers: • Selection facilitated by DNA markers can improve efficlency of plant breeding, par'.IculaJ~ly when selection is undertaken in early generations or where |election is difficult, time consuming or lX~nliv@. Two genii, one for roils=ante to black root rot, the other for resistance to PVY", ~ere selected for screening for linkage to RAPD markers. 3.I ~ RAPD M%~B~ GonoTated by P~imer 83 ~as Ti=htlv Linkqd, Ln ReD~I~OD. to che mUFfS7 49 Gene for Resistance ~o Black Root Ro~: The single dominant gone for black root rot resistance was originally from ~urley 49, an A~aricantobacco variety, and was transferred in=o Canadian ~oDacco varieties by backcross. The rloulting lane was designated PBIg. NOaE isogonic lines w~th the gone in a Delgold background have been developed during the past I0 years. A total of 150 DNA samples were extracted from Delgold, PBIg, one neat: isogonic line, and F: plants from the original PBlg/Delgold cross. The same plants were tested for black root ro~ resistance. Initially, 200 primers were screened for amplification of polymorphic DNA fragmencs on the genomio DNA isolated frc~ Delgold (without ~ho gone) and the near isogonic line in Delgold (with the gone). The random primers, which gave rise to polymorl~hi= RAPD markers, were fuz'cher tested with genomi¢ DNA from the ~ plants segregating for resistance to black roo~ rot° One RAPD marker (aJ~out 500 bp) was found to be tightly linked, in repulsion, to the target gins. Screening for additional ~ markers using another 200 random primers is under way. it is hoped ~haU a marker linked in coupling can be deter"cod. 3.2 PvY Resistance Go~@: resistance in NC744, an Rmerican breeding line, is controlled by a single recessive geneo Douoc~ion of RAPD marklrl aseocia~edwi~h ~he glne is essential because screening for resistance with pVY virus is no~ allowed aU Delhi Rases=oh S~aUion. Secondly phenotypi¢ detection of ~he resistance mus~ be done on self progeny because of the recessive nature of ~he gone; therefore i~ takes more generations to transfer the gone into a Canadian ~obaooo variety using conventional backcrosaing. RAPD ~aJ~ker- assisted selection for resistance to pVY virus will overcome the r !imitation. one hundred and fifty DNA samples wore ox~ a¢~od from Dolgcld, NC744, and F: plants derived from the cross NC744/Delgold. The same plants were shipped ~o Vineland Research S~ation for the evaluation of resistance ~o PVY". Resistance screening is near comple~icn and the experiment should be completed by March of 1994. 4. .~APD Finaer~rints of Canadian Tobacco Variet£esz ;nttial screening for RAPD markers using 50 random primers and glnomic DNA from the Canadian varieties, Delhi 34, Delhi 76, Delgold, Canda~, Dallier and Delfie~d, indicated very low levels of P.~PD polymo~hiam. The pa~terns of the RAPD fingerprints of Canadian varieties were also similar ~o those of American varieties However, ~he pat~o=nl of ~he RAPD fingerprints of ~he Eastern Europe ~obaoco varieties were much different .:~ om those of ~he Noah Rmerioan varieties. Screening with more randnm primers is needed before the RAPD fingerprin~ for each of the Canadian uobacco varieties is established. DNA can also be ex~rac~id from cured =:bocce and used in fingerprinting should the need arise. CD P~ w Ln P~ CO B.A.T. CONFIDE'NT[AL - CATEGOP.Y h NIINNESOTA TOBACCO LITIGATION BATCo document for PFSFC 1 March 1999

- 40 - Se Field Evaluation of B~ Lines Derived *- __ Somali= HvbEids: Nine BC: lines derived f:omN.~ (+) ~. svlvestrin som4=ic hyb:ids WQ=I ~littd ~n i fiild ~=i&l :hii ~11: laE. FOU: final Wi:h y~lld and ~ldt l:dix ec~al to Dol~old weEt Ldln=ifitd a~td 8~qll plKZl~l HIOC~ fz'om =hoau, i lineo aro w3w being ~ToJmmd ~ a diallol pa~ern Co to~ tho S~ I ~nazcation. T~oo Co fo~hu~ Syn ~ p~a..'l~s will be ~m,'n in ~ho ~leLd nex~ yeL: and sing~e p~an~ selections taken fo= se~l~; and b&Gkc~oil~ng to Dil~old. ~olicl:lon pEeilu:e will be L~DoI~ on ~lm s~lgle Syn Z plamts to ol~Lr~Co noz-nico~nm convocCm:s f:om the poG~la~ion. 6. N. r~a~£ca ~+I n. ~abac~m ~vb~ids: The thl:d backc:osu ~o Del~old h~s been comp~ot~ ~nd seed stocks rill be ovaluaC~d to do~erm~ne £¢ 8ufflcienc stad hli ~eon pz-oduced co conduce • field Cost in 1994. 0 ~J B.A.T. CONFIDENTIAL - CATEGORY I: MINNESOTA TOBACCO LITIGATION BATCo document for PFSFC 1 March 1999

- 41 - PROCZSS DZVF.LO¥1G~T r~ B.A.T. CONFIDENTIAL - CATEGORY I: MINNESOTA TOBACCO LITIGATION BATCo document for PFSFC 1 March 1999

- 42 - RECORD TYPE: SUB TYPX: SECURITY CODE, FUNDING BODY: ORGANZY, ATZON: GROUP NUMBER- LOCAL PROJECT NUMRER(S): PROJECT TITLE; PERSON RESPONSIBLE: EFFORT: PROJECT DESCRIPTION: SCOPE: DEPTH = FUNCTION= OB,,TECTZ',~: ~-..,USTER: DATE REVIEW WRITTEN: REVT~W TITLE: REV~L~ TEXT: ITL CANADA 448 T-S349;G6 Lamina Process OpClmlzaCion HIRTLE, K. To ¢~:l:~are our d~-ying process 1=o ~AT~,'7*S HTD, EATUKE°8 improved ITM and Hauni'| HT in order to solo~: the bes~ potential got ~he future, f=r filling value optimization. LOCAL APPLIED/DEVELOPMZNT COST PRODUCT RES/9EV January 1994 Lamina Drying Optimization A ~hree-way T.es~ was conduced in ¢:~peE&~ion wiC.~ BATCT, BATUKE and RADii personnel, using varioum combinauions of we~Uln~/s~eaming and drying meU~od8. Experimental tobacco samples have been analyzed and cigarettes ma~ufac=ured. Results indicate tha~ BATCF's RTD process produced ~slght reductions of up to 22% s= equal fLT~ess an~ 11% a~ equal draw resistance, It and 0t for BATUK~'s and 7% and It for Hauni's, rsspe~Zivoly. It has boon derided to s~ay wi~h ~he currsn= rotary dzTs= process since Chore is no Inusnticn ~o eliminate DZ~T in our cu=rsn= successful brands by fuzZ:he= expandLng ~he base materla~. O B.A.T. CONFIDENTIAL - CATEGORY h MINNESOTA TOBACCO LITIGATION BATCo document for PFSFC t March 1999

- 43 - I~ECOR~ TYPE: SUB TYPE: SECURITY CODE: FUNDING BODY: ¢RGkNIZATZON: GR~FP NUMBER: LCX::~$, PROJECT hUMBER ( $ ) : PROJECT TITLE, PERSON RESPONSIBLE: EFFORT: PROTECT DESCRZPTION: SCOPE: DEPTH: FUNCTION: OS/EC~ZVE .- CLUSTER: P S ITL CANADA 456 T-S350-1;L-13;06 Stem Poetess 0pcimizac£on - STS Process HIRTLE, K. To Qcudy B.A.T. Germany's $TS (Steam T=ea~ed Stem) expansion process re= stem. To evaluate STS unit An an ITL plant. To investigate the effo¢~ of all s~em process paramm~ern in order to reduce filling value variaciona and to increase drying capacity. To provide new process specifications for all plants if quali~y or c081: improvemmn~ can be made. LOCAL APPLIED/DEVELOPMENT PRODUCTION PHYSICAL PROCESS RES/DEV DATE REV~EWWRITTEN= January 1994 REVIEW TITLE: Stem Process Optimization - STS Poetess. REVIEW TEXT: ~TS ~£nimql Expansion STS having @quivalenu fill value UO WTS was subjectively =c~npa=ed ~o WTS in two of ITL'e major brands. Reeul~e indicated tha~ negative ~aste differences do ~mllt wi~h STS. Thl8 comparison was done wi~h STS having IS0 cp£. Samples p~paEed a~ 135 cpi have been assessed by an external product tee~ panel, differences have been found on one brand. PuSher investigation is rmquizld. CD m B.A.T. CONFIDENTIAL - CATEGORY I: i~IINNESOTA TOBACCO LITIGATION BATCo document for PFSFC 1 March 1999

- 44 - RECORD TYPE: SUB TYPE= SZCuR'r "1"~ COD~": FUNDING BODY: ORG~I Z~T2ON: GROUP NUMBER: LOCAL PROJECT NUMBER{S): pROJECT TITLE: PERSON RESPONSIBLE: EFFORT: PROJECT DZSCR~PT~ON: SCOPE: DEPTH: FUNCTION: OBJECTIVE: CLUSTER: DAT~ REVIEW WRITTEN: REV;EW TITLE" REVIEW TEXT: a) (~q: L£ne Sereenina: aseessmen~ of recent mod£ficatlons; ITL CANkDA 448 T-5352;06 GLT Process Optimization HIRTLE, K., I.EVE.RT. G. Optimization of Green Leaf Threshing P~anU processes nhrough replacement and Improvements to existing l,;uil~e~t. Phase Zl of a machinery/process upgrading program is be£ng completed. Phase I91 (stems) is being planned. LOCAL APPLIED/DEVELOPMZNT LEAF COST PRODUCT RES/DEV January 1994 GLT ProCess OpclmlzatiQn V~LEIOUS process studies a~e being carried ouc: ~p~ovmncs Co the sand reel and Co ~he screen configuration. ~q~-drvsr O~allficacion: qualificaCion is being completed with a chE0ugh~ut of 29,000 ibe./hr, at 700 grains. c} Cond£uionin=: mo£sture meter evaluation, ol~:imiza~ion of operation. Separator Lcadinu: C~imizanlon. e) Balker Size/Slicer OD~imiza=ion. f) Mac Vac Pre-condltioner O~l~niza~icn. Sale Feed System Evaluetlon. h) ?orq~qn Material De~ecti~n: SLmco-Rauni= System evaluation. i} Dus~.Co~c~ors S~een£nu Imurovemsn~. CD w ~n B.A.T. CONFIDENTIAL - CATEGORY I: ~IINNESOTA TOBACCO LITIGATION BATCo document for PFSFC 1 March 1999

- 45 - RECORD TYPE: SO3 TYPE: SECURITY CODE: FUND ZI~ BODY: ORG,~iZZ~TION: GROUP h'UHBER: LOCAL PROJECT h'U~(S] : FRO, TECT TITLE: P~OZ( I~SPONSIRX.E: F.I~fORT: PROJECT DESCRII=TION : SCOPE: DEPTH= FUNCTION: CLU~= DATE REVIEW WRITTEM= REVIEW TITLE: TEXT: P S ZTL CANADA 448 T-6541;06 P:o:~ec~ Co~t: .. R~NFR.~, C. Suppo~ thm intEoduc~ian of PCL-X at manufa~-~uring plant level. LOCAL EPPLIED/DEVELOPMENT PRODUCTION COST ~COUCT I~S /DEV January 1994 Project Comet All ¢£garatte brands now toni&in PCL-X. P~L-X fah=icaCion pcocoss ks being optimized so that packed moisture contont can be increased from 9.5 Co 10.5%. (This will :educe lumps at ¢IgaEecue planes without excemslvm dus~ geneEation). CD r~ B.A.T. CONFIDENTIAL - CATEGORY I: MINNESOTA TOBACCO LITIGATION BATCo document for PFSFC 1 March 1999

- 46 - WORK AREA 07 R G D SUPPORT SERVICZS C L-- B.A.T. CONFIDENT[AL ° CATEGORY I: NIINNESOTA TOBACCO LITIGATION BATCo document for PFSFC 1 March 1999

- 47 - RECORD TYPE: SUB TYP[: SECURITY CODE: FUND THG BODY: ORGANI ZATION: GROUP ~(BER - LO(:m.L PROJECT ~'MSER[S): PROJECT TITLE: PERSON RESPONSIBLE: EFFORT: PROJECT DESCRIPTION: SCOPE: DEPTH: FUNCTI ON: OBu'ECTIVE : CLUSTER: DATE REVIEW WRITTEN= REVZZW TITLE : REVTE~ TEXT: ITL CANADA 410 T-S010; T-5000; T-5679; 07 Evaluation of Hew Package Films DUNN.R This week is aimed at providing options towards improving the shelf llfe of ZTL products. The evaluations consider two avenues: "chose films which may have harrier prope~ies equlvalen~ to current films but whose be=net sealing characteristics facilitate manufacturing$ those films which have at ~he onset improved moisture barrier properties. This work is linked to other activities under ~he "snvlronmen=al umbrella'. GROUP APPLIED/DEVELOPMENT January 1994 Evaluation of New Package Film" A number of films have been screened in the laboratory and under very shor~ runs in plant conditions. Testing included evaluation of physical characteristics, potential for ink lift and sealability, package seal quality (CPST) and molsturt ~ten~ion index (accelerated lhmlf-life ~es~}. On of the films used by ITL is the Hercules SXT 80 gauge film with a moisture vapour transmission rate (MVTR) of 0.34 g/100 in2/day at 100"F, 90% Following preliminary tests, the follow£ng two films are considered of greatest interest: a) GLS 20 1~lypro~ylene film from Z.C.I. has a somewhat higher MVTR =hat Hercules 8XT (0.39 vs 0.34 g/1C0 in:/24 hours/0.8 mil) bur conslstsn~ly be~tsr package sealing has increased moisture retention index by approximately 10%. A build-in shrink chara~eristic for tigh~ wrapping has been found to cause unwind tracking problems when nearing the 90 mm 0.D. core. A trial nn recommended 120 mm O.D. cores Improved funning. larger ~=ial is pending. b) Hercules X0-19 poIyp~pylene film with a quoted MVTR of 0.21 for 80 gauge provided an increase An moisture retention index of 35-40%. Running trials have been satisfactory and the film has been qualified. Preliminary ~esting of other regular and "hlgh-harrisE" films are expected ~o be £nitiated within the ne~c few months. CD B.A.T. CONFIDENTIAL - CATEGORY I: ~[INNESOTA TOBACCO LITIGATION BATCo document for PFSFC 1 March 1999

0 r~ BATCo document for PFSFC 1 March 1999