Unusually high levels of carcinogenic tobacco-specific nitrosamines in Sudan snuff (toombak)

Abstract

The aim of these studies was to determine the levels of carcinogenic tobacco specific nitrosamines (TSNA) in Sudanese oral snuff (toombak) as recent retrospective epidemiological studies suggested an association between the use of toombak and subsequent development of oral cancer. We have analyzed the TSNA levels in 20 samples of Sudanese toombak, of four different quality levels, collected from five different vendors. Using GC coupled with thermal energy analysis, four TSNA were quantified in snuff extracts: N'-nitrosonornicotine (NNN), N'-nitrosoanatabine (NAT), N'-nitrosoanabasine (NAB) and 4-(methyl-nitrosamino)-1-(3-pyridyl)-1-butanone (NNK). Unusually high levels of these TSNA (mean; range, mg/g snuff, dry wt) were detected; NNN (1.13; 0.50-3.08); NAT (0.08; 0.02-0.29); NAB (0.22; 0.02-2.37); and NNK (2.31; 0.62-7.87). Previously, the highest levels of NNN and NNK reported in any snuff were 0.154 and 0.014 mg/g dry wt respectively. In comparison, the levels in Sudanese toombak were up to 20 and 560 times higher respectively. As the public health implications of these findings are significant, attempts should be made to reduce exposure to TSNA in oral snuff users in Sudan.

The Swedish Snus and the Sudanese Toombak: are they different?

“…and N H -nitrosoanabasine (NAB) were quanti®ed in toombak and saliva of toombak users [15,16]. Exceptionally high levels (mean; range, mg/g toombak/dry wt) of NNN (1.13; 0.50±3.08), NNK (2.31; 0.62±7.87; 31), NAT (0.08; 0.02±0.2) and NAB (0.22; 0.02±2.37) were found (Table 1).…”

“…Much later, it was found that 81% (50/62) of patients with oral squamous cell carcinoma (SCC) from the Sudan used toombak [36]. Recently, abundant information on toombak use and development of oral cancer has been reported, and the cancer was frequently found at the site where the toombak quid is placed [9,12,15,16,21]. Studies of a similar nature from Saudi Arabia have suggested the aetiological role of Shamma for development of oral cancer [37,38].…”

Differences in the Bacteriome of Smokeless Tobacco Products with Different Oral Carcinogenicity: Compositional and Predicted Functional Analysis

“…The American, Sudanese and Yemeni products had comparable bacterial loads. However, studies indicate that they significantly vary with respect to their TSNA content [39]; SuT in particular has very high concentrations of TSNAs [40]. Obviously then, the levels of TSNAs in ST products differ not only as function of total microbial load but probably also as a function of microbial community composition.…”

“…A standard curve was generated by running 10-fold serial dilutions of E. coli DH10B genomic DNA. Amplification was carried on a ViiA7 platform (Applied Biosystems, USA) including an initial enzyme activation cycle at 95°C for of 10 min followed by 40 (AGAGTTTGATYMTGGCTCAG) [24] and 519R (GWATTACCGCGGCKGCTG) [25], modified to contain Illumina's specific adapter sequences 803F and 1392wR, were used to amplify the V1-3 region of the 16S rRNA gene using standard PCR conditions. The resultant PCR amplicons (~ 520 bp) were then purified (Agencourt AMPure XP beads, Beckman Coulter, USA), indexed with unique 8-base barcodes (Nextera XT v2 Index Kit sets A-D, Illumina, USA) and pooled together in equimolar concentrations.…”

Snuff Dipping Sudanese Tombak Keratosis - A Case Report

“…The nitrose amines levels that found in Tobacco are high in toombak [10], attributed to the type of tobacco Species and way of processing toombak at high temperature, prolonged storage, and contamination during processing [11][12][13].…”