title: In vitro cytotoxicity of Nicotiana gossei leaves, used in the Australian Aboriginal smokeless tobacco known as pituri or mingkulpa.
abstract: The Aboriginal population of Central Australia use endemic Nicotiana species to make a smokeless tobacco product known usually as pituri or mingkulpa. Nicotiana leaves are masticated with wood ash into a 'quid' that is chewed/sucked for absorption of nicotine. In addition to nicotine, smokeless tobacco products contain a spectrum of biologically active compounds that may contribute to effects on health. The objective of this study was to quantify nicotine, and related alkaloids and tobacco specific nitrosamines (TSNAs), in Nicotiana leaves used in pituri, and compare in vitro toxicity of pure nicotine with Nicotiana leaf extract at the same concentration of nicotine. An aqueous extract of dry leaves of Nicotiana gossei and a reference smokeless tobacco (CORESTA CRP2) were quantified for major pyridine alkaloids and TSNAs using HPLC-UV and LC-MS/MS. A range of extract concentrations and corresponding concentrations of nicotine standard were tested using an MTS assay to measure human lung epithelium cell (A549) survival. Cells treated for 24h with the maximum concentration of 1.5mg/ml of nicotine resulted in 77% viability. In contrast, extracts from N. gossei leaves and CRP2 containing a similar concentration of nicotine (1.3mg/ml) resulted in remarkably lower viability of 1.5 and 6%, respectively. Comparison of cytotoxicity of pure nicotine with that of the extracts revealed that nicotine was not the source of their cytotoxicity. Other biologically active compounds such as the known carcinogens NNK and NNN, derived from nicotine and nornicotine and found to be present in the smokeless tobacco extracts, may be responsible.
authors.affiliation: School of Pharmacy, The University of Queensland, Brisbane, Qld 4072, Australia.
authors.forename: Nahid
authors.initials: N
authors.lastname: Moghbel
authors.affiliation: School of Pharmacy, The University of Queensland, Brisbane, Qld 4072, Australia.
authors.forename: BoMi
authors.initials: B
authors.lastname: Ryu
authors.affiliation: School of Pharmacy, The University of Queensland, Brisbane, Qld 4072, Australia.
authors.forename: Peter J
authors.initials: PJ
authors.lastname: Cabot
authors.forename: Angela
authors.lastname: Ratsch
authors.affiliation: School of Pharmacy, The University of Queensland, Brisbane, Qld 4072, Australia. Electronic address: k.steadman@uq.edu.au.
authors.forename: Kathryn J
authors.initials: KJ
authors.lastname: Steadman
mesh_terms: D045744:Cell Line, Tumor; D002470:Cell Survival / Q000187:drug effects; D002851:Chromatography, High Pressure Liquid; D004305:Dose-Response Relationship, Drug; D004847:Epithelial Cells / Q000187:drug effects* / Q000473:pathology; D006801:Humans; D008168:Lung / Q000187:drug effects* / Q000473:pathology; D009538:Nicotine / Q000302:isolation & purification / Q000633:toxicity; D009602:Nitrosamines / Q000302:isolation & purification / Q000633:toxicity; D010936:Plant Extracts / Q000302:isolation & purification / Q000633:toxicity*; D018515:Plant Leaves; D013056:Spectrophotometry, Ultraviolet; D053719:Tandem Mass Spectrometry; D014030:Tobacco, Smokeless / Q000633:toxicity*
chemical_list: D009602:Nitrosamines; D010936:Plant Extracts; D009538:Nicotine; C016583:4-(N-methyl-N-nitrosamino)-1-(3-pyridyl)-1-butanone; C008655:N'-nitrosonornicotine
keywords: A549 lung carcinoma cell line; Alkaloid; Cytotoxicity; Mingkulpa; Nicotiana gossei; Nicotine; Pituri; TSNA; Tobacco-specific nitrosamines