Toxicologic evaluation of flavor ingredients added to cigarette tobacco: skin painting bioassay of cigarette smoke condensate in SENCAR mice.
Four comparative two-stage SENCAR mouse skin painting bioassays were conducted with cigarette smoke condensate (CSC) preparations to evaluate the effect of common American cigarette flavoring ingredients on tumor promotion. Each independent study employed a unique flavoring combination applied to tobacco at exaggerated levels, and in total resulted in an evaluation of 150 ingredients. Groups of 30-50 female SENCAR mice each were initiated topically with 50 microg of 7,12-dimethylbenz(a)anthracene (DMBA), and promoted thrice weekly for 26 weeks with either 10 or 20 mg of CSC from test cigarettes containing ingredient mixtures. For comparison, separate groups of mice received concurrent treatment with CSC from reference cigarettes prepared without added ingredients. Negative and positive controls were treated with acetone or 12-0-tetradecanoyl-phorbol-13-acetate (TPA) as a promoter, respectively. CSC-only groups served as promotion controls. Tumors developed in > 80% of the TPA-treated mice by study week 11, with a < 3% background tumor formation in the acetone treated controls at termination. Tumor incidence in CSC-only promotion control groups was < 20%, with no apparent difference between reference and test CSC groups. Approximately 70% of the DMBA-initiated mice promoted with 20 mg CSC developed tumors. Tumors first appeared around week 9, with about five tumors/tumor bearing animal. Tumor incidence, latency and multiplicity were CSC dose related, with a lower tumor incidence (approximately 50%), longer latency (12 weeks), and reduced tumor burden (four tumors/tumor bearing animal) at the 10 mg CSC dose level. While tumor incidence, latency and multiplicity data occasionally differed between test and comparative reference CSC groups, all effects appeared to be within normal variation for the model system. Furthermore, none of the changes appeared to be substantial enough to conclude that the tumor promotion capacity of CSC obtained from cigarettes containing tobacco with ingredients was discernibly different from the CSC obtained from reference cigarettes containing tobacco processed without ingredients.
Gaworski
C L
CL
Lorillard Tobacco Co., Greensboro, NC 27420, USA. cgaworski@lortobco.com
Heck
J D
JD
Bennett
M B
MB
Wenk
M L
ML
D015127:9,10-Dimethyl-1,2-benzanthracene / Q000032:analysis / Q000633:toxicity; D000287:Administration, Topical; D000818:Animals; D001681:Biological Assay; D001835:Body Weight / Q000187:drug effects; D002248:Carbon Monoxide / Q000633:toxicity; D002273:Carcinogens / Q000032:analysis / Q000633:toxicity; D005260:Female; D005421:Flavoring Agents / Q000633:toxicity*; D051379:Mice; D018632:Mice, Inbred SENCAR; D009538:Nicotine / Q000008:administration & dosage / Q000633:toxicity; D018722:Nicotinic Agonists / Q000008:administration & dosage / Q000633:toxicity; D010947:Plants, Toxic; D012867:Skin / Q000187:drug effects* / Q000473:pathology; D012878:Skin Neoplasms / Q000139:chemically induced / Q000473:pathology; D012906:Smoke / Q000009:adverse effects* / Q000032:analysis*; D012907:Smoking; D016019:Survival Analysis; D013638:Tars / Q000633:toxicity; D014026:Nicotiana / Q000737:chemistry*
D002273:Carcinogens; D005421:Flavoring Agents; D018722:Nicotinic Agonists; D012906:Smoke; D013638:Tars; C024746:tobacco tar; D015127:9,10-Dimethyl-1,2-benzanthracene; D009538:Nicotine; D002248:Carbon Monoxide