Tobacco Documents Online

. CHEMICAL STUDIES ON'TOBACCO SMOKE LXVI. ASSESSI•SENT OF VOLATILE AND TOBACCO-SPECIFDC'N-NITROSAMINES IN THE SMOKE OF'SOME SELECTED WEST GERMAN CIGARETTES1. C. Ri9h1 Abteilung ft5r Toxikologie, Max-von-Pettenkofer Institut, Bundesgesundheitsamt, Berlin, F.R.G. John D. Adams and Dietrich Hoffmann Naylor Dana Institute for Disease Prevention American Heal'th Foundation Valhalla, New York 10595 U.S.A. 1This study was supporte&by NCI Contract No.1-CP-55666 and by the Federal Ministry for Youth, Family & Health of the Federal Republic of Germany.

J ° The prc•sence of both volatile t:-nitres:amincs (Vt:T;) and • tobacco-s_uecific nonvolatilie h-nitrosamii:es (NVNA)' in cigar.ette smoke (1,2) and their selective reduction in mainstream smoke by cellulose acetate filter tips has been reported previously (3-5). The levels of VNA in undiluted:sidestream smoke (SS) exceed those in mainstream smoke (MS) up to several hundred times. The concentration oi VNA in SS, however, depends greatly on the air fl'ow around the burning cone (4). Under standard con- ditions the total amount of NVNA generatediin SS is comparable with that produced in the MS. So far, these observations were only reported for popular U.S. cigarettes and recently for the VNa of some commercial cigarettes from the U.K. (6). In this study we analyzed the smoke of some West German cigarettes for VNA and NVNA. Material and Methods. Apparatus. For the MS analysis we used a 20-port automatic_ smoker (H. Borgwaldt, Hamburg, Germany) with a rotating head where every second port is connected to a nitrogen source, thus replacing the air in the trap with nitrogen every 2 seconds (7). For the SS analysi~s we utiliized a single-channel smoker (H. Borgwaldt, Hamburg, Germany) and SS apparatus ad- justed to an air flow of 1500 m1/min (8). For one particular filter cigarette we studied the effects of flow rates of 250, 500, 1000 0 and 1,500 ml/min on 1 the yields of VNA in the SS.

\ For the VNA analysis we employed a Hewlett-Packard Model 700 gas liquid' chromatograph int.erfaced with a thermal energy analyzer (TEA; Thermal Electron Corp., Waltham, Mass.) and a (Hewlett-Packard Modei, 3380:A) recording integrator. For the analysis of NVNA in the final concentrate we used for the HPLC-separation a Model 6000 A solvent delivery system (Waters Associates, Inc., Milford, Mass.) with a Model' 70-101sample injection valve equipped with a Model 70-11 loop filller port (Rheo- dyne, Berkely, Calif.) and Corasil II and 2 v-Porasil columns (Waters Associates, Inc.). The TEA with HPLC in- terfaced (Thermo Electron Corp., Waltham, Mass.) with a Hewlett-Packard Model 3380 A recording integrator served as detector. In addition, smoke of one brand each of filter an&nonfilter cigarettes was also analyzed for individual N-nitrosamines with a Hewlett-Packard ModeL 5982 A GLC-MS instrument. A Nuclear Chicago Isocap 300 was used for scintillation counting in toluene with 0!.5$ PPO and 0.0058 POPOP. Reagents. [14CjN-Nitrosodimethylamine (14C-NDMA): 0.48 mCi/ mmol) was employed as an~internal standard for VNA and (2,-14C]N'-ni~trosonornicotine (14'.1 mCi/mmol; 9) for NVNA. ~. The VNA standards were kindly provided by Dr. T. Fazio, ~ Food and Drug Administration, Washington, D.C. N'-Nitroso- ~ . r N - N - 2 . W

noynicotine (NNN), 4-(h'-methyl-N-nitrosami~no)-1-(3-pyridyl)-1- bnta;ione (t:?:7{) and N'-nitrosoanatabine (NAtB) were synthesized by methods reported in the literature (2,9,10). All solvents used throughout this study were free from VNA and NVNA accord- i~ng to GLC-TEA analysis. Cigarettes. All cigarettes for VNA analyses (except Cigarette. B) were purchased on the open market in Berlin during 1978. Cigarette B and all other brands for the NVNA analysis were bought in Berlin in 1979. The cigarettes were stored'in a humidity chamber at 22°+2° at relative humidity of 60+3% for 24 hours and were selected to be within +20 mg of the average. weight of 200 cigarettes. The cigarettes were smoked in•a tobacco laboratory maintained at a relative humidity of 60+5% and at 22'°+2°. Throughout the study we applied'standard smoking conditions taking 1 puff/min of 2 second duration_. and 35 m1i volume, and smoking to 23 mm butt length. For filter cigarettes the butt length was determined by the length of the filiter plus+ overwrap plus 3 mm (11). [These smoking conditions are identical with those of the Deutsche Normen DIN 10240 for machine smoking of cigarettes (12)]. Mainstream Smoke Analysis. The procedures have been described in detail in earlier publications (2,4),. In Figure 1, we have sum.~narized'the various steps of the analysis for VNA and in Figure 2 the procedure for NVNA. - 3 -

Sidestream Smoke Analysis. For the SS analysis 6 cigarettes selected for weight and draw resistance, were smoked in the special apparatus (Figure 3). The analytical procedure been described earlier (4). Results. has Figure 4 shows a gas chromatogram of volatile N-nitrosamines in the mainstream smoke of a popular West German cigarette. In addition to the major VNA, N-nitrosodimethylamine (NDMA), N-nitrosoethylmethylamine (NEMA) and N-nitrosopyrroliidine- (NPYFt)i, GLC-MS analysis showed also the presence of N-nitroso- diethylamine and N-nitrosopiperidine (<1' ng/cig). Table l lists the results for NDMA, NEMA and NPYR in NLS and SS of 2 popular nonfilter cigarettes and 4 popular cellulose acetate fliliter cigarettes from the West German market. Unexpectedly, the MS contained higher levels of NPYR than of NDMA. Generally, these data confirmed the earlier finding that the VNA are significantly lower in the MS of cellulose acetate filter cigarettes than in the MS of nonfilter cigarettes because of selective filtration (2,3). Under standard smoking conditions the VNA concentrations were much greater in SS than in MS. However, variation of the - e: -

air flow rates during smoking, al~ters the MS/SS ratios of Vti'A, as presented in Table 2. The flow rate of 1,500 ml per minute through the sidestream smoke apparatus was determined to reflect the standard condition, because it yielded data for TPM, nicotine and VNA in the mainstream smoke eamparable to a cigarette scmking,in the opan air (Figure 3). In model experi- ments with 14C-labelled markers, we are currently studying the basic mechanisms that lead to the formation of VNA in MS and SS, and:we explore possibilities for the reduction of VNA in SS. Figure 5 depicts a high performance liquid chromatogram of the NVNA in the MS of a West German cigarette and Table 3 lists the quantitative data calculated from the response of the thermal energy analyzer as a detector for tobacco specific N-nitrosamines in the MS of 6 selected West German cigarettes. It appears that these values are at about the same levels as are those for NVNA in U.S. cigarettes (2). We are currently studying the correlation of nicotine in tobacco, Nox in smoke and yield of NVNA in the mainstream smoke and we are examining various filters for their capability to selectively remove the tobacco-specific N-nitrosamines (5). - 5 -

Table 1. VOLATILE N-NITROSnf•fINES IN MAINSTREAM SMOKE (MS) AND SIDESTREAM Sr;OKE (SS) OF SOME POPULAR 1•SEST GERMAN CIGARETTES1 Brand Type Smoke NDMA ng/cig NEMA ng/cig NPYR ag/cig A (20;1.2) NF MS 13.8 1.1 30.3 SS 514. 311. 510. B (22;11.3) NF MS 6.8 ND 11.5 SS 213. 115.4 281. C (14;0.7) F MS 3.1 ND 8.4 SS 330. 13.2 296. D (112;0.7) F' MS 1.8 ND 5.7 SS 558. 35. 700. E (13; 01.7) F MS 2.4 ND 3.1 SS 408. 20. 333. F (10 ; 0 . 4 ) F. MS 2.2 ND 8.7 SS 538. 30. 500. 1A11, values are averages of 2 analyses; the cigarettes were 85 mm long; numbers in parentheses represent mg values of TPM and nicotine, respectively. ND- not determined (<1 ng/cig). NF = nonfilter cigarette F = filter cigarette

Table 2. CO.ti'CEN"1'('.i.'DIO.J OF TaREE VOLATILE N-NITROSAhiINES IN SIDES'PREAAi SMOKE GENERATED UNDER DIFFERENT AIR FLOW'RATES1 CIGARETTE C Flow Rate ml/min NDMA ng/cig NENIA ng/cig NPYR ng/cig 1,500 330 13.2' 296 1,000 272 11.4 226 500 185 9.8~ 189 250 128 6.7' 106 1Average of 2 runs

Table 3. NONVOLATILE N-NITROSAAfi*dES IN FLAII:STR£F;M SIAOKE OF SOME POPULAR WEST GER:-4A'`I CIGARe.TTESl Brand Type NAtB ('ug/cig 1 NNN t Lg/cl'g 1 NNK - (hg/cig) A NF 0.19 0.51, 0.08 B NF 0'.07 0.23 0.04 C F 01.17 0.22 0.06 D F 0.04 0.112 0.02 E F 0.07 0.18 0.05 F F 0.07 0.18 0.06 cigarettes were 85 mm long. lAll values are averages of 2 analyses; the NF = nonfilter cigarette F = filter cigarette

Referenccs 1. Schmeltz, I., and Hoffmann, Ds: Chem. Rev. 77 (1977) 295-311. 2. Hoffmann, D., Adams, J.D., Brunnemann, K.D., and Hecht, S.S.: Cancer Res. 39 (1979) 2505-2509. 3. Morie, G.P., and Slloan, C.H.: Beitr. Tabakforsch. 7(1973) 61-66. - 4. Brunnemann, K.D., Yu, L., and Hoffmann D.: Cancer Res. 37 (1977) 3218-3222. 5. Piade, J.J., Adams, J!.D., and Hoffmann, D.: Abstr. 33rd Tobacco G1emists'Res. Conf. Lexington, Kentucky, October 29- 31, 1979 p.31. 6. Annual Report, 1978, Laboratory of Government Chemist, 7. London, U.K. _ Hoffmann, D., Hecht, S.S., Schmeltz, I., Brunnemann,, K.D., and Wynder, E.L.: Recent Advan. Tobacco Sci. 1(19'76): 97-122. 8. Brunnemann, K.D., and Hoffmann, D.: Food Cosmet. Toxicol_ 12 (1974) 115-124. 9. Hu, M.W'., Bondinell, W.E., and Hoffmann, D.: J. Labelled Compd. 10 (1974) 79-88. 10. Hecht, S.S., Chen, C.B., Hirota, N., Ornaf, R.M., Tso, T.C., and Hoffmann, D.: J. Natl. Cancer Inst. 60 (1978) 819-824. 11. Bates, W.W., Griffith, R.B'., Harlow, H.S., Senkus, W., and Wakeham, H.: Tobacco Sci. 12 (1968) 192-196. 12. Deutsches Institut ftir Normung e.V. DIN 1024D, Part 1, (Febr. 1977) 3 pp'-