Abstract

Introduction: Toombak is a smokeless tobacco used by the Sudanese. Nicotiana Rustica leaves are fermented, and sodium bicarbonate is added, increasing Ph. This causes high absorption spikes of unprotonated nicotine through mucosal epithelium, and into the blood, distinguishing Toombak as an addictive product. Furthermore,Toombak hashighmicrobial contamination due to its open field, rural production. Materials and methods: 21 tobacco samples were collected from 3 main towns in the capital Khartoum, Sudan. Analysis included microbiome (16S rRNA sequencing), metabolome (Liquid and Gas Chromotography- volatile organic compound method), heavy metal content and scanning electron microscopy (SEM). Results: Compared to limit of detection values (LOD), Tobacco specific nitrosamines (TSNA’s), in particular, 4(methylnitrosamino)-1-(3-pyridyl)–butanone (NNK) were markedly increased [NNK; 1.2-4.7 mg/g; LOD 0.02]. Free nicotine ranged from 16-31 mg/g; [LOD 0.01]. High choline and carnitine, volatile aldehydes; and benzyl alcohol were also detected. 8 cations, of which iron [1465 mg/kg] and copper [3.76 mg/kg] were strikingly observed, compared to average levels from other products; [Iron 20-60mg/kg, Copper 1.3mg/kg]. 8 Phyla, including but not limited to; Actinobacteria; cornyebacterium(contain nitrate reductase genes) and Firmicutes; staphylococcus and facklamia, were sequenced. SEM highlighted a non-homogenous product with elevated sodium spectrum. Conclusions: TSNA’s were potent in Toombak due to high starting nicotine and rich nitrate reductase bacteria. High choline and carnitine can promote cardiovascular disease through their conversion to trimethylamine-N-oxide. Diphtheria and infective endocarditis are associated with cornyebacterium and facklamia respectively while staphylococcus can lead to numerous systemic opportunistic diseases.

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