Assessment and comparison of the levels of N-nitrosonornicotine and 4-(n-methyl-n â€“nitrosamino) -1-(3-pyridyl)-1-butanone in the saliva of tobacco chewers and non- chewers -a hospital based study
Keywords:NNK, NNN, Saliva, Smokeless Tobacco, Tobacco-Specific Nitrosamines (TSNAâ€™s).
Background: Studies estimating the Tobacco- specific nitrosamines, (TSNAâ€™s) which are the strongest carcinogens in the saliva of
tobacco users and tobacco quitters, are limited.
Objectives: To assess and compare the levels of N- nitrosamines (NNN, NNK) in the saliva of tobacco chewers and non -chewers
including those who have quit the habit of tobacco use.
Methods: The study included 120 patients who were divided into three groups of 40 each: Group I- Smokeless tobacco chewers
Group II- Tobacco chewers who have completely stopped the habit at least 2 weeks prior to sample collection and
Group III- non-chewers. The salivary levels of two tobacco specific nitrosamines; NNN & NNK levels were estimated in the three study groups. Statistical analysis was done by Kruskalâ€“ Wallis, one-way analysis of variance (ANOVA) test, Mann-Whitney U test. (p-value < 0.05 was considered to be statistically significant)
Results: In Group I, the mean level of NNN was 651.84 ± 359.78 and mean level of NNK was 168.32 ± 131.83. In Group II, the mean level of NNN was 119.52 ± 95.05 and mean level of NNK was 42.78 ± 43.19. In Group III, the mean level of NNN was 3.44 ±6.55 and mean level of NNK was 1.98 ± 3.68. There was a statistical difference in the 3 groups with respect to mean levels of NNN and NNK.
Conclusion: The study indicated that salivary tobacco-specific nitrosamines are elevated in tobacco chewers. Saliva can be used to detect TSNAâ€™s and screen for TSNAâ€™s during each patientâ€™s de-addiction process.
 International Institute for Population Sciences (IIPS), Ministry of Health, and Family Welfare (MoHFW). Global adult tobacco survey (GATS) India 2009â€“2010. Mumbai and New Delhi: IIPS and MoHFW, Government of India; 2010.
 Hoffmann D, Adams JD. Carcinogenic Tobacco-specific N-Nitrosamines in Snuff and in the Saliva of Snuff Dippers. Cancer Research 1981; 41: 43054308.
 Sharma Sugandha, Rajesh K Joshi et al. â€œModified UFLC-PDA Method for determination of Nitrosaminesâ€ Journal of Analytical Chemistry September 2015, Volume 70, Issue 9 , pp 1153 â€“ 7
 IARC. Monographs on the evaluation of carcinogenic risks to humans. Betel- quid and areca nut chewing and some related nitrosamines.vol. 85. Lyon: IARC, 2004.
 Stepanov I, Jensen J, Hatsukami D, Hecht SS. Tobacco-specific nitrosamines in new tobacco products. Nicotine Tob Res. 2006 Apr; 8(2):309-13. https://doi.org/10.1080/14622200500490151.
 Brunnemann KD, Hornby AP, Stich HF. Tobacco-specific nitrosamines in the saliva of Inuit snuff dippers in the Northwest Territories of Canada. Cancer Lett. 1987 Oct; 37(1):7-16. https://doi.org/10.1016/0304-3835(87)90140-6.
 Bhide SV, Nair UJ, Nair J, Spiegelhalder B, Preussmann R. N-nitrosamines in the saliva of tobacco chewers or masheri users. Food Chem Toxicol. 1986 Apr; 24(4):293-7. https://doi.org/10.1016/0278-6915(86)90005-0.
 Shah N, Sharma PP Role of chewing and smoking habits in the etiology of oral submucous fibrosis (OSF): a case-control study J Oral Pathol Med 1998; 27: 475â€“9 https://doi.org/10.1111/j.1600-0714.1998.tb01915.x.
 Patil P, Bathi R, Chaudhari S et al. Prevalence of Oral mucosal lesions in dental patients with tobacco smoking, chewing, and mixed habits : A cross-sectional study in South India J. Family Community Med.2013 May- Aug; 20(2) :130-5 https://doi.org/10.4103/2230-8229.114777.
 Joshi Urvish, Modi Bhavesh et al. A study on prevalence of chewing form of tobacco and existing quitting patterns in Urban population of Jamnagar, Gujarat Indian J Community Med.2010 Jan;35(1):105-8.
 Global Adult Tobacco Survey (GTAS) Indonesia report https://doi.org/10.4103/0970-0218.62560.
 Sinha DN, Gupta PC Tobacco and areca nut use in male medical students of Patna. National Medical Journal of India. 2001; 14:176â€“8.
 Global Adult Tobacco Survey (GTAS-2) India 2016-2017.
 Tobacco Institute of India Fact sheets Universal Leaf Tobacco; UN Trade database (2013).
 Goniewicz Maciej, Eisner Mark, Ponce Eduardo et al. Comparison of Urine Cotinine and the Tobacco-Specific Nitrosamine Metabolite 4- (Methylnitrosamino)-1-(3-Pyridyl)-1-Butanol (NNAL) and Their Ratio to Discriminate Active From Passive Smoking Nicotine & Tobacco Research, Volume 13, Number 3 (March 2011) 202â€“8.
 Lakritz L, Gates R.A., Gugger A.M et al. Nitrosamines levels in human blood, urine and gastric aspirate following ingestion of foods containing potential Nitrosamine precursors or preformed Nitrosamines Fd. Chem Toxic Vol.20, pp 455 to 459, 1982.
 Stepanov I, Jensen J, Hatsukami D et al. Tobacco â€“specific Nitrosamines in new tobacco products Nicotine and Tobacco Research Volume 8, Number 2 (April 2006) 309-13 https://doi.org/10.1080/14622200500490151.
 Dominique Kavvadias, Gerhard Scherer, Francis Cheung, Graham Errington et al. Determination of tobacco-specific N-nitrosamines in urine of smokers and non-smokers Biomarkers, 2009; 14(8): 547â€“53.
 Hecht SS, Carmella SG, Stepanov I, Jensen J, Anderson A, Hatsukami DK. Metabolism of the tobacco-specific carcinogen 4-(methylnitrosamino)-1-(3- pyridyl)-1-butanone to its biomarker total NNAL in smokeless tobacco users. Cancer Epidemiol Biomarkers Prev. 2008 Mar; 17(3):732-5. https://doi.org/10.1158/1055-9965.EPI-07-2843.
 Yoshizawa J, Schafer C, Schafer J et al. Salivary Biomarkers: Toward Future Clinical and Diagnostic Utilities Clin Microbiol Rev. 2013 October; 26(4): 781â€“91.
 Hamzany Y, Savulescu D, Iris Borovoi et al. Salivary analysis of Oral cancer biomarkers British Journal of Cancer (2009) 101, 1194 â€“ 8 https://doi.org/10.1038/sj.bjc.6605290.
 Jaiswal P, Goel A, Mittal R.D Survivin : A molecular biomarker in cancer Indian J Med Res. 2015 Apr; 141(4): 389-97 https://doi.org/10.4103/0971-5916.159250.
 Santolia D, Gupta S, Mahajan B et al. Salivary expression of Inerleukin-6 in Oral potentially malignant disorders and Oral squamous cell carcinoma patients International Journal of Current Research Vol.8, Issue 11,pp 42443-42446, November 2016.
 Kulza M, Wozniak A, Przybylowska SM, Czarnywojtek A, Kurhanska-Flisykowska A, Florek E. Saliva cotinine determination using high-performance liquid chromatography with diode - array detection. Przeglad Lekarski Journal. 2012;69(10):837â€“40.
 V Asha, M Dhanya Immunochromatographic Assessment of salivary cotinine and its correlation with Nicotine dependence in tobacco chewers J Cancer Prev. 2015 Jun; 20(2): 159-63 https://doi.org/10.15430/JCP.2015.20.2.159.