Assessment of Trace Elements in Commercially Packaged Forms of Tobacco Using Laser-induced Breakdown Spectroscopy

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K., Adarsh U. K. This is a preprint; it has not been peer reviewed by a journal. https://doi.org/ 10.21203/rs.3.rs-2695953/v1 This work is licensed under a CC BY 4.0 License Status: Posted Version 1 posted You are reading this latest preprint version Abstract BACKGROUND: Oral cancer is the most common malignancy in many developing countries such as India due to increased consumption of smokeless tobacco. Elemental components in commercially packaged forms of tobacco can play a significant role in the causation of oral cancer. AIM: Assess trace elements in various types of commercially packaged forms of tobacco using LASER-induced breakdown spectroscopy. MATERIALS AND METHODS: Two types of ‘Paan masala’ and four types of packaged tobacco were obtained from local ‘paan’ shops. The contents in the packets were made into pellets respectively using a hydraulic press and subjected to elemental analysis using Laser-induced breakdown spectroscopy. RESULTS : A ten-trial experiment was carried out in all six pellets. The National Institute of Standards and Design (NIST) database was used to assess the emission lines. The elements obtained in both commercially packaged tobacco and ‘paan masala’ were similar. The elements are Calcium, Iron, Aluminium, Nickel, and Chromium. CONCLUSION: It is known that substances that cause DNA damage and carcinogenesis, are inorganic elements such as nickel and Chromium. It is clear in our study that these carcinogens are present in the commercially packaged form of tobacco and ‘paan masala’ samples. oral cancer smokeless tobacco trace elements Laser-Induced breakdown spectroscopy Figures Figure 1 Figure 2 Figure 3 Figure 4 Figure 5 Figure 6 Figure 7 Figure 8 Figure 9 Figure 10 Introduction It is a well-established fact that tobacco in both smokeless and smoked forms has a major role to play in causing oral potentially malignant disorders (OPMDs) and oral cancer ( 1 ). “Smokeless tobacco implies the use of unburned tobacco in the finished products that are sucked, chewed (dipped), gargled, or applied to the gums or teeth, while fine tobacco mixtures are usually inhaled into the nostrils” ( 2 ). It is also often referred to as “spit” or “spitting” tobacco as the chewers spit the saliva and the extracts of tobacco that build up in the oral cavity. This smokeless tobacco usually contains nicotine, tobacco, abrasives, chemicals, sweeteners, and salts. These habits influence oral health and lead to the risk of oropharyngeal carcinoma ( 3 ). The use of smokeless tobacco along with or without betel quid is common in many south Asian countries. Betel quid is “a substance, or mixture of substances, placed in the mouth or chewed and remaining in contact with mucosa (for prolonged periods of time) usually containing at least one of the two basic ingredients, tobacco or areca nut, in raw or any manufactured or processed form”( 4 )( 5 ). Prolonged exposure to these products causes micromolecular alteration, which is often followed by morphological alteration in the oral mucosa that is clinically evident as OPMDs which later may have the potential to turn into oral cancer ( 6 ). On the other hand, ‘paan masala’ is a “dehydrated non-perishable preparation of areca nut, catechu (Acacia catechu). slaked lime (Calcium oxide and Calcium hydroxide), cardamom, and many artificial perfuming and flavoring substances” ( 7 ). ‘Paan masala’ is often substituted for smokeless tobacco as it claims to contain no tobacco-related products. The predisposing changes in the oral mucosa of oral cancer are referred to as OPMDs and trace elements play a major role in causing OPMDs ( 8 ). Trace elements are broadly classified as macro elements (Ca, P, Mg Na, K, Cl, and S) and microelements (Fe, Cu, I, Mn, Zn, Mo, Co, F, Se, and Cr). These trace elements such as Magnesium, Zinc, Manganese, Cobalt, and Iron are involved in defence mechanisms owing to their antioxidant properties ( 9 ). While, Chromium, Nickel, Cadmium, Arsenic, and Lead are considered carcinogens ( 10 ). Considering, OPMDs statistically increase the risk of progressing into malignancy and in India, 7% of lesions with severe epithelial dysplasia undergo malignant transformation ( 11 ). There is a need to assess the presence of trace elements in the commercially packaged form of tobacco and ‘paan masala’ to understand the role of trace elements in the causation of OPMDs and oral cancer. Our objective was to assess trace elements in commercially packaged tobacco and ‘paan masala’ using Laser Induced Breakdown Spectroscopy (LIBS). Materials And Method Sample collection: A total of four commercially packaged forms of tobacco and two ‘paan masala’ packets were obtained from the local ‘paan’ shops in Mangalore, India. The list of the ingredients as described by the manufacturers was noted. ‘Paan masala’ packets mentioned the presence of no tobacco and their ingredients were areca nuts, catechu, lime, permitted spices, cardamom, lime, menthol, and added flavours. Commercially packaged forms of tobacco mentioned presence of 100% tobacco. The sample name of each ‘paan masala’ and commercially packaged form of tobacco (for the purpose of the study) were named as PM1, PM2, T1, T2, T3, and T4 respectively (Figure1). Sample preparation: The contents in the packet were in a semi-powdered form. The contents of the packet were grounded into an even powder using a mortar and pestle. 2g of the powder was weighed and made into a pellet using a hydraulic press. The formed pellet (Figure 2) was further dried using a desiccator. The sample thus obtained was subjected to Laser-induced Breakdown spectroscopy (LIBS). The LIBS system uses a high-energy pulsed laser (Nd: YAG) operating at 532 nm with 6 ns pulse width, exposure time of 10 ms, and 10 Hz repetition rate to ablate the sample. The laser beam was reflected in the orthogonal direction using a glass prism, followed by a biconvex lens (lens 1) with a 6 cm focal length, which focuses the beam to a micrometer-size spot. The high-energy laser beam focused on the sample surface creates sufficient energy density to induce ablation of the sample in a localized spot, and the plasma is created. Plasma emits the characteristic emissions of its constituents which are collected and collimated by the same lens (lens 1) used for focusing the laser. The collimated signal is then focused on to the spectrograph input by another biconvex lens (lens 2) of focal length 20 cm. Spectrograph grating disperses the signal in the wavelength domain, which is then recorded using a CCD detector connected to a PC. The recorded spectra from samples are subjected to data pre-processing including baseline correction, normalization, etc. It is followed by analysis based on the known-element database (local and NIST database) to qualitatively or quantitatively characterize the samples. The pellet was mounted and subjected to a 10-trial experiment. The pellet was exposed to a single shot of the laser pulse in different regions. Results Two different samples of ‘paan masala’ and four different samples of commercially packaged forms of tobacco were labelled as PM1, PM2, T1, T2, T3, and T4 respectively. The ten-trial experiment was carried out in the spectral range of 260 nm – 520 nm. The emission lines of Calcium were perceived to be the most significant in ten different wavelengths specifically 316.17 nm, 370.71 nm, 393.75 nm 396.26 nm, 422.68 nm, 430.53 nm, 431.03 nm, 432.15 nm 442.76 nm, and 445.75 nm. The emission lines of Iron were perceived to be more significant in nine different wavelengths specifically 285.56 nm, 318.19 nm, 344.05 nm, 358.39 nm, 360.16 nm, 373.72 nm, 400.04 nm, 404.47 nm, and 438.64 nm. The emission line of Aluminium was perceived in two different wavelengths specifically 279.86 nm and 309.59 nm. The emission line of Nickel was perceived significantly in a single wavelength specifically 280.6 nm. The emission line of Chromium was perceived significantly in a single wavelength specifically 357.52 nm. On comparing ‘Paan masala’ which claimed no tobacco and commercially packaged tobacco (100% tobacco), there was no difference in elemental composition of both the subgroups. Hence the spectral range of both the sub-groups was plotted in a single graph as shown in Figure 3 The average intensity of the most intense emission lines of different elements with their standard deviation of PM1, PM2, T1, T2, T3, and T4 are listed in Table 1 – 6. Table 1 Average intensity of trace elements present in PM1. ELEMENTS MOST INTENSE LINE (nm) AVERAGE INTENSITY (counts) Al 279.86 Ni 280.62 Cr 357.51 Fe 374.97 Ca 393.88 Table 2 Average intensity of trace elements present in PM2. ELEMENTS MOST INTENSE LINE (nm) AVERAGE INTENSITY (counts) Al 279.86 Ni 280.62 Cr 357.51 Fe 374.97 Ca 393.88 Table 3 Average intensity of trace elements present in T1. ELEMENTS MOST INTENSE LINE (nm) AVERAGE INTENSITY (counts) Al 279.86 Ni 280.62 Cr 357.51 Fe 374.97 Ca 393.88 Table 4 Average intensity of trace elements present in T2. ELEMENTS MOST INTENSE LINE (nm) AVERAGE INTENSITY (counts) Al 279.86 Ni 280.62 Cr 357.51 Fe 374.97 Ca 393.88 Table 5 Average intensity of trace elements present in T3. ELEMENTS MOST INTENSE LINE (nm) AVERAGE INTENSITY (counts) Al 279.86 Ni 280.62 Cr 357.51 1014 Fe 374.97 802.85 Ca 393.88 Table 6 Average intensity of trace elements present in T4. ELEMENTS MOST INTENSE LINE (nm) AVERAGE INTENSITY (counts) Al 279.86 Ni 280.62 Cr 357.51 Fe 374.97 Ca 393.88 The intensity of Calcium was the highest and Chromium was least in PM1 (i.e., Ca>Al>Ni>Fe>Cr) as shown in Figure 4. The intensity of Calcium was the highest and iron was least in PM2 (i.e., Ca> Cr> Al>Ni>Fe) as shown in Figure 5. The intensity of Calcium was the highest and Chromium was least in T1 (i.e., Ca > Al> Ni> Fe> Cr) as shown in Figure 6. The intensity of Calcium was the highest and Chromium was least in T2 (i.e., Ca > Al> Ni> Fe> Cr) as shown in Figure 7. The intensity of Calcium was the highest and iron was least in T3 (i.e., Ca > Al> Ni>Cr> Fe) as shown in Figure 8. The intensity of Calcium was the highest and iron was least in T4 (i.e., Ca > Al> Ni>Cr> Fe) as shown in Figure 9. Comparison of trace elements (Al, Fe, Ca, Ni, and Cr) in commercially packaged tobacco The average intensities of the trace elements (Al, Fe, Cr, Ni, and Ca,) in all six groups of ‘paan masala’ and commercially packaged form of tobacco were compared using the ANOVA Kruskal Wallis test (listed in Table 7). There was statistically significant difference between the concentration of trace elements in the six different samples of ‘paan masala’ and commercially packaged tobacco (p <0.001). The median and Interquartile range was higher in Ca. We can draw the inference that the concentration of Calcium is highest and least Iron in packaged tobacco, then the level of Aluminium and then the level of Nickel and then Chromium as shown in Fig 10 (Ca>Al>Ni>Cr>Fe). Table 7 Kruskal Wallis test to compare the level of trace elements in commercially packaged tobacco ELEMENTS INTENSITY MEDIAN INTERQUARTILE RANGE P VALUE Al 2552.62 4351.17 <0.001 Ni 2877.39 1154.99 1768.37 2237.31 <0.001 Cr 905.83 1931.84 <0.001 Fe 923.26 717.54 <0.001 Ca 39818.45 36306.05 <0.001 Discussion Our study aimed to assess elements in commercially packaged forms of tobacco and ‘paan masala’ that claimed to have no tobacco. To the best of our knowledge Laser-induced breakdown spectroscopy (LIBS) has not been employed in assessing the elemental composition of commercially packaged tobacco and ‘paan masala’. Since LIBs is a “put and play” method, it has been used for various applications such as elemental mapping of trace elements in different hard and soft tissue leading to early diagnosis of malignancies, calculi/ stones, biological or dental material like pellets or powders, and detection of bacteria/ viruses in bioaerosols (12)(13). LIBS aids in the multifaceted detection of elements and does not require sample preparation. Furthermore, it is a noncontact experimental procedure, and all states of matter such as solid, liquid, and gas can be assessed (14). It is a well-established fact that smokeless tobacco products have a major role to play in causing OPMDs and oral cancer(7). Chiba, et al (15) reviewed various toxic elements in tobacco and smokeless tobacco. They showed the presence of various elements in tobacco products such as Cr, Al, Ni As, Mn, Cu, Pb, Hg, and Zn. Our study showed a similar presence of trace elements that were listed by the authors. The concentration of Aluminium and Nickel were relatively higher and lesser in Chromium. Some trace elements are chemical carcinogens which may be genotoxic or non-genotoxic. Genotoxic carcinogens interact directly with the DNA resulting in chromosomal aberration and DNA damage. While, the latter are chemicals that act directly as tumor promotors, induce inflammatory response, cause immunosuppression, and tissue toxicity. Cr, Be, Ni, Cd, and As are considered carcinogens by International Agency for Research on Cancer (IARC) (8). Nickel initiates carcinogenesis by attaching to DNA protein causing mutilated DNA leading to cross-linking of DNA interstrand and DNA protein, and breakage of the DNA strand. Chromium initiates carcinogenesis by producing reactive oxygen species (ROS) and suppressing p53 gene (8) (16). We also observed highest concentration level of Calcium and lowest concentration of Iron in commercially packaged tobacco and ‘paan masala’ samples. Dhaware, et al (17) determined the presence of toxic metals such as As, Cd, Cu, and Pb in various types of Indian smokeless tobacco (gutkha, creamy stuff, khaini, mishri, tooth powder, zarda) products using differential pulse anodic stripping voltammetry (DPASV). There were also negligible concentrations of other elements such as Ni, Cr, and Hg. Mohammed et al (18) quantified the presence of various elements in Shisha and Doksha tobacco products using Energy Dispersive X – ray Fluorescence (EDXRF). They observed higher concentrations of Calcium, and other elements such as Al, Ni, Cr, Fe, Mg, K, Sr, Zn, and Mg. Our study also showed, all the commercially packaged forms of tobacco have a higher concentration of Calcium compared to other elements and presence of carcinogenic elements such as Ni, Cr, and other elements such as Fe, and Al was also observed. Patients having the habit of chewing various smokeless tobacco, and smoking tobacco products with confirmed OSMF were assessed to understand the role of trace elements in the etiopathogenesis of OSMF by Bagewasi, et al (19) and Kode, et al (20). They also stated that there was an elevation of salivary Copper levels in patients having OSMF with the habit of chewing various tobacco products. There was also alteration in Cu/Zn ratio and decreased Fe levels in patients having OSMF with habits. Surprisingly, there was no difference in the elemental composition of commercially packaged tobacco and ‘paan masala’ packets (that claimed no tobacco) in our study. Conclusion Consumption of smokeless forms of tobacco has increased for the past one decade in India and in southeast Asian regions. Packaged forms of tobacco and betel quid are popularly consumed in India. Our study revealed the presence of various elements such as Calcium, Aluminum, Iron, and known carcinogenic elements such as Nickel and Chromium. Interestingly commercially packaged form (Paan masala), claimed by the manufacturer as tobacco free has similar composition as that of tobacco containing packets. We recommend studying elemental composition of body fluids such as blood, saliva and tissue specimen in patients with deleterious chewing habits. We hope such a study will give us insight into the role of elements in etiopathogenesis of OPMDs and oral cancer. Declarations Acknowledgement The authors are thankful for the support rendered by the Device Development Program, Department of Science & Technology (DST), Government of India (DST/TDT/DDP-26/2018) and Department of Atomic Energy (DAE), Board of Research in Nuclear Sciences (BRNS), Government of India (34/14/04/2014-BRNS). References Vini Mary Antony J, Ramani P, Anuja, Sherlin HJ, Jayaraj G, Don, et al. Detection of trace metals among the most commonly available smoked and smokeless tobacco products. J Pharm Sci Res. 2017;9(10):1855–7. Group IA for R on CW. Personal habits and indoor combustions. Volume 100 E, A review of human carcinogens. IARC monographs on the evaluation of carcinogenic risks to humans. 2009; Waheed S, Siddique N, Rahman S. 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Bagewadi SB, Hirpara DR, Paliwal A, Raiyani BD, Hafiz A, Vasra HD, et al. Estimation of Salivary Copper, Zinc, Iron, and Copper-to-zinc Ratio in Oral Submucous Fibrosis Patients : A Case – control Study. 2022;2–5. Ankolekar Kode M, Rashmiraj Karjodkar F. Estimation of the serum and the salivary trace elements in OSMF patients. J Clin Diagnostic Res. 2013;7(6):1215–8. Additional Declarations No competing interests reported. Cite Share Download PDF Status: Posted Version 1 posted You are reading this latest preprint version Research Square lets you share your work early, gain feedback from the community, and start making changes to your manuscript prior to peer review in a journal. As a division of Research Square Company, we’re committed to making research communication faster, fairer, and more useful. We do this by developing innovative software and high quality services for the global research community. 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Also discoverable on Platform About Our Team In Review Editorial Policies Advisory Board Help Center Resources Author Services Accessibility API Access RSS feed Manage Cookie Preferences © Research Square 2026 | ISSN 2693-5015 (online) Privacy Policy Terms of Service Do Not Sell My Personal Information {"props":{"pageProps":{"initialData":{"identity":"rs-2695953","acceptedTermsAndConditions":true,"allowDirectSubmit":true,"archivedVersions":[],"articleType":"Research Article","associatedPublications":[],"authors":[{"id":183871368,"identity":"993c4856-87b3-46b5-a329-ec730f1e6171","order_by":0,"name":"Kripa Adlene Edith A","email":"","orcid":"","institution":"Manipal College of Dental Sciences, Mangalore","correspondingAuthor":false,"submittingAuthor":false,"prefix":"","firstName":"Kripa","middleName":"Adlene Edith","lastName":"A","suffix":""},{"id":183871369,"identity":"15f193ad-334c-4d2b-b2b9-58674988bd67","order_by":1,"name":"Ravikiran Ongole","email":"data:image/png;base64,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","orcid":"","institution":"Manipal College of Dental Sciences, Mangalore","correspondingAuthor":true,"submittingAuthor":false,"prefix":"","firstName":"Ravikiran","middleName":"","lastName":"Ongole","suffix":""},{"id":183871370,"identity":"a8672fdf-805c-49a3-bb42-0df77586dfbb","order_by":2,"name":"Unnikrishnan V. 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sample\u003c/p\u003e","description":"","filename":"5.png","url":"https://assets-eu.researchsquare.com/files/rs-2695953/v1/fac5aa50850ad104006e1be5.png"},{"id":34548785,"identity":"1f9bd199-48bc-428f-8af8-eea5ae61f96e","added_by":"auto","created_at":"2023-03-20 22:08:36","extension":"png","order_by":6,"title":"Figure 6","display":"","copyAsset":false,"role":"figure","size":14252,"visible":true,"origin":"","legend":"\u003cp\u003eThe average intensity of all the trace elements in the T1 sample\u003c/p\u003e","description":"","filename":"6.png","url":"https://assets-eu.researchsquare.com/files/rs-2695953/v1/bb3fbc116e16fb84245dd98b.png"},{"id":34548790,"identity":"fa038bbe-14bd-487c-8803-af39316989cb","added_by":"auto","created_at":"2023-03-20 22:08:36","extension":"png","order_by":7,"title":"Figure 7","display":"","copyAsset":false,"role":"figure","size":15195,"visible":true,"origin":"","legend":"\u003cp\u003eThe average intensity of all the trace elements in the T2 sample\u003c/p\u003e","description":"","filename":"7.png","url":"https://assets-eu.researchsquare.com/files/rs-2695953/v1/ad960298928254e948cb6f32.png"},{"id":34548791,"identity":"7c7267b6-b51e-4a3b-b55f-87c304841d20","added_by":"auto","created_at":"2023-03-20 22:08:36","extension":"png","order_by":8,"title":"Figure 8","display":"","copyAsset":false,"role":"figure","size":16749,"visible":true,"origin":"","legend":"\u003cp\u003eThe average intensity of all the trace elements in the T3 sample\u003c/p\u003e","description":"","filename":"8.png","url":"https://assets-eu.researchsquare.com/files/rs-2695953/v1/2acf704044e9dfab29d4534d.png"},{"id":34549680,"identity":"f3689f32-f818-476d-8791-b0453d2b508b","added_by":"auto","created_at":"2023-03-20 22:16:36","extension":"png","order_by":9,"title":"Figure 9","display":"","copyAsset":false,"role":"figure","size":16229,"visible":true,"origin":"","legend":"\u003cp\u003eThe average intensity of all the trace elements in the T4 sample\u003c/p\u003e","description":"","filename":"9.png","url":"https://assets-eu.researchsquare.com/files/rs-2695953/v1/762c94b92a2eeddc93c9d981.png"},{"id":34548787,"identity":"50ab5df1-b91c-4bd7-99ee-1ac98e645357","added_by":"auto","created_at":"2023-03-20 22:08:36","extension":"png","order_by":10,"title":"Figure 10","display":"","copyAsset":false,"role":"figure","size":18642,"visible":true,"origin":"","legend":"\u003cp\u003eThe graph shows the level of trace elements in commercially packaged tobacco.\u003c/p\u003e","description":"","filename":"10.png","url":"https://assets-eu.researchsquare.com/files/rs-2695953/v1/373f469772b8bc28031a20c6.png"},{"id":35220798,"identity":"0394b97d-8ede-4bde-ad7a-6184e4447e4e","added_by":"auto","created_at":"2023-04-03 18:44:21","extension":"pdf","order_by":0,"title":"","display":"","copyAsset":false,"role":"manuscript-pdf","size":707813,"visible":true,"origin":"","legend":"","description":"","filename":"manuscript.pdf","url":"https://assets-eu.researchsquare.com/files/rs-2695953/v1/c4ddf23c-9247-4d07-abbd-479dbf86e9b2.pdf"}],"financialInterests":"No competing interests reported.","formattedTitle":"\u003cp\u003eAssessment of Trace Elements in Commercially Packaged Forms of Tobacco Using Laser-induced Breakdown Spectroscopy\u003c/p\u003e","fulltext":[{"header":"Introduction","content":"\u003cp\u003eIt is a well-established fact that tobacco in both smokeless and smoked forms has a major role to play in causing oral potentially malignant disorders (OPMDs) and oral cancer (\u003cspan citationid=\"CR1\" class=\"CitationRef\"\u003e1\u003c/span\u003e). \u0026ldquo;Smokeless tobacco implies the use of unburned tobacco in the finished products that are sucked, chewed (dipped), gargled, or applied to the gums or teeth, while fine tobacco mixtures are usually inhaled into the nostrils\u0026rdquo; (\u003cspan citationid=\"CR2\" class=\"CitationRef\"\u003e2\u003c/span\u003e). It is also often referred to as \u0026ldquo;spit\u0026rdquo; or \u0026ldquo;spitting\u0026rdquo; tobacco as the chewers spit the saliva and the extracts of tobacco that build up in the oral cavity. This smokeless tobacco usually contains nicotine, tobacco, abrasives, chemicals, sweeteners, and salts. These habits influence oral health and lead to the risk of oropharyngeal carcinoma (\u003cspan citationid=\"CR3\" class=\"CitationRef\"\u003e3\u003c/span\u003e). The use of smokeless tobacco along with or without betel quid is common in many south Asian countries. Betel quid is \u0026ldquo;a substance, or mixture of substances, placed in the mouth or chewed and remaining in contact with mucosa (for prolonged periods of time) usually containing at least one of the two basic ingredients, tobacco or areca nut, in raw or any manufactured or processed form\u0026rdquo;(\u003cspan citationid=\"CR4\" class=\"CitationRef\"\u003e4\u003c/span\u003e)(\u003cspan citationid=\"CR5\" class=\"CitationRef\"\u003e5\u003c/span\u003e). Prolonged exposure to these products causes micromolecular alteration, which is often followed by morphological alteration in the oral mucosa that is clinically evident as OPMDs which later may have the potential to turn into oral cancer (\u003cspan citationid=\"CR6\" class=\"CitationRef\"\u003e6\u003c/span\u003e). On the other hand, \u0026lsquo;paan masala\u0026rsquo; is a \u0026ldquo;dehydrated non-perishable preparation of areca nut, catechu (Acacia catechu). slaked lime (Calcium oxide and Calcium hydroxide), cardamom, and many artificial perfuming and flavoring substances\u0026rdquo; (\u003cspan citationid=\"CR7\" class=\"CitationRef\"\u003e7\u003c/span\u003e). \u0026lsquo;Paan masala\u0026rsquo; is often substituted for smokeless tobacco as it claims to contain no tobacco-related products. The predisposing changes in the oral mucosa of oral cancer are referred to as OPMDs and trace elements play a major role in causing OPMDs (\u003cspan citationid=\"CR8\" class=\"CitationRef\"\u003e8\u003c/span\u003e). Trace elements are broadly classified as macro elements (Ca, P, Mg Na, K, Cl, and S) and microelements (Fe, Cu, I, Mn, Zn, Mo, Co, F, Se, and Cr). These trace elements such as Magnesium, Zinc, Manganese, Cobalt, and Iron are involved in defence mechanisms owing to their antioxidant properties (\u003cspan citationid=\"CR9\" class=\"CitationRef\"\u003e9\u003c/span\u003e). While, Chromium, Nickel, Cadmium, Arsenic, and Lead are considered carcinogens (\u003cspan citationid=\"CR10\" class=\"CitationRef\"\u003e10\u003c/span\u003e). Considering, OPMDs statistically increase the risk of progressing into malignancy and in India, 7% of lesions with severe epithelial dysplasia undergo malignant transformation (\u003cspan citationid=\"CR11\" class=\"CitationRef\"\u003e11\u003c/span\u003e). There is a need to assess the presence of trace elements in the commercially packaged form of tobacco and \u0026lsquo;paan masala\u0026rsquo; to understand the role of trace elements in the causation of OPMDs and oral cancer. Our objective was to assess trace elements in commercially packaged tobacco and \u0026lsquo;paan masala\u0026rsquo; using Laser Induced Breakdown Spectroscopy (LIBS).\u003c/p\u003e"},{"header":"Materials And Method","content":"\u003cp\u003e\u003cstrong\u003e\u003cem\u003eSample collection:\u003c/em\u003e\u003c/strong\u003e\u003cem\u003e\u0026nbsp;\u003c/em\u003eA total of four commercially packaged forms of tobacco and two \u0026lsquo;paan masala\u0026rsquo; packets were obtained from the local \u0026lsquo;paan\u0026rsquo; shops in Mangalore, India. The list of the ingredients as described by the manufacturers was noted. \u0026lsquo;Paan masala\u0026rsquo; packets mentioned the presence of no tobacco and their ingredients were areca nuts, catechu, lime, permitted spices, cardamom, lime, menthol, and added flavours. Commercially packaged forms of tobacco mentioned presence of 100% tobacco. \u0026nbsp; The sample name of each \u0026lsquo;paan masala\u0026rsquo; and commercially packaged form of tobacco (for the purpose of the study) were named as PM1, PM2, T1, T2, T3, and T4 respectively (Figure1).\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003e\u003cem\u003eSample preparation:\u0026nbsp;\u003c/em\u003e\u003c/strong\u003eThe contents in the packet were in a semi-powdered form. The contents of the packet were grounded into an even powder using a mortar and pestle. 2g of the powder was weighed and made into a pellet using a hydraulic press. The formed pellet (Figure 2) was further dried using a desiccator. The sample thus obtained was subjected to Laser-induced Breakdown spectroscopy (LIBS).\u003c/p\u003e\n\u003cp\u003eThe LIBS system uses a high-energy pulsed laser (Nd: YAG) operating at 532 nm with 6 ns pulse width, exposure time of 10 ms, and 10 Hz repetition rate to ablate the sample. The laser beam was reflected in the orthogonal direction using a glass prism, followed by a biconvex lens (lens 1) with a 6 cm focal length, which focuses the beam to a micrometer-size spot. The high-energy laser beam focused on the sample surface creates sufficient energy density to induce ablation of the sample in a localized spot, and the plasma is created. Plasma emits the characteristic emissions of its constituents which are collected and collimated by the same lens (lens 1) used for focusing the laser. The collimated signal is then focused on to the spectrograph input by another biconvex lens (lens 2) of focal length 20 cm. Spectrograph grating disperses the signal in the wavelength domain, which is then recorded using a CCD detector connected to a PC. The recorded spectra from samples are subjected to data pre-processing including baseline correction, normalization, etc. It is followed by analysis based on the known-element database (local and NIST database) to qualitatively or quantitatively characterize the samples. The pellet was mounted and subjected to a 10-trial experiment. The pellet was exposed to a single shot of the laser pulse in different regions.\u003c/p\u003e"},{"header":"Results","content":"\u003cp\u003eTwo different samples of \u0026lsquo;paan masala\u0026rsquo; and four different samples of commercially packaged forms of tobacco were labelled as PM1, PM2, T1, T2, T3, and T4 respectively. The ten-trial experiment was carried out in the spectral range of 260 nm \u0026ndash; 520 nm. The emission lines of Calcium were perceived to be the most significant in ten different wavelengths specifically 316.17 nm, 370.71 nm, 393.75 nm 396.26 nm, 422.68 nm, 430.53 nm, 431.03 nm, 432.15 nm 442.76 nm, and 445.75 nm. The emission lines of Iron were perceived to be more significant in nine different wavelengths specifically 285.56 nm, 318.19 nm, 344.05 nm, 358.39 nm, 360.16 nm, 373.72 nm, 400.04 nm, 404.47 nm, and 438.64 nm. The emission line of Aluminium was perceived in two different wavelengths specifically 279.86 nm and 309.59 nm. The emission line of Nickel was perceived significantly in a single wavelength specifically 280.6 nm. The emission line of Chromium was perceived significantly in a single wavelength specifically 357.52 nm.\u003c/p\u003e\n\u003cp\u003eOn comparing \u0026lsquo;Paan masala\u0026rsquo; which claimed no tobacco and commercially packaged tobacco (100% tobacco), there was no difference in elemental composition of both the subgroups. Hence the spectral range of both the sub-groups was plotted in a single graph as shown in Figure 3\u003c/p\u003e\n\u003cp\u003eThe average intensity of the most intense emission lines of different elements with their standard deviation of PM1, PM2, T1, T2, T3, and T4 are listed in Table 1 \u0026ndash; 6.\u003c/p\u003e\n\u003ctable border=\"1\" cellpadding=\"0\" cellspacing=\"0\" width=\"636\"\u003e\n \u003ctbody\u003e\n \u003ctr\u003e\n \u003ctd colspan=\"3\" valign=\"top\" width=\"100%\"\u003e\n \u003cp\u003e\u003cstrong\u003eTable 1\u0026nbsp;\u003c/strong\u003e\u003cstrong\u003eAverage intensity of trace elements present in PM1.\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" width=\"22.20472440944882%\"\u003e\n \u003cp\u003eELEMENTS\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"32.75590551181102%\"\u003e\n \u003cp\u003eMOST INTENSE LINE (nm)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"45.039370078740156%\"\u003e\n \u003cp\u003eAVERAGE INTENSITY (counts)\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" width=\"22.20472440944882%\"\u003e\n \u003cp\u003eAl\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"32.75590551181102%\"\u003e\n \u003cp\u003e279.86\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"45.039370078740156%\"\u003e\n \u003cp\u003e\u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" width=\"22.20472440944882%\"\u003e\n \u003cp\u003eNi\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"32.75590551181102%\"\u003e\n \u003cp\u003e280.62\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"45.039370078740156%\"\u003e\n \u003cp\u003e\u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" width=\"22.20472440944882%\"\u003e\n \u003cp\u003eCr\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"32.75590551181102%\"\u003e\n \u003cp\u003e357.51\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"45.039370078740156%\"\u003e\n \u003cp\u003e\u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" width=\"22.20472440944882%\"\u003e\n \u003cp\u003eFe\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"32.75590551181102%\"\u003e\n \u003cp\u003e374.97\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"45.039370078740156%\"\u003e\n \u003cp\u003e\u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" width=\"22.20472440944882%\"\u003e\n \u003cp\u003eCa\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"32.75590551181102%\"\u003e\n \u003cp\u003e393.88\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"45.039370078740156%\"\u003e\n \u003cp\u003e\u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003c/tbody\u003e\n\u003c/table\u003e\n\u003cp\u003e\u0026nbsp;\u003c/p\u003e\n\u003ctable border=\"1\" cellpadding=\"0\" cellspacing=\"0\" width=\"633\"\u003e\n \u003ctbody\u003e\n \u003ctr\u003e\n \u003ctd colspan=\"3\" valign=\"top\" width=\"100%\"\u003e\n \u003cp\u003e\u003cstrong\u003eTable 2\u0026nbsp;\u003c/strong\u003e\u003cstrong\u003eAverage intensity of trace elements present in PM2.\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" width=\"32.859399684044234%\"\u003e\n \u003cp\u003eELEMENTS\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"32.859399684044234%\"\u003e\n \u003cp\u003eMOST INTENSE LINE (nm)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"34.28120063191153%\"\u003e\n \u003cp\u003eAVERAGE INTENSITY (counts)\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" width=\"32.859399684044234%\"\u003e\n \u003cp\u003eAl\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"32.859399684044234%\"\u003e\n \u003cp\u003e279.86\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"34.28120063191153%\"\u003e\n \u003cp\u003e\u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" width=\"32.859399684044234%\"\u003e\n \u003cp\u003eNi\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"32.859399684044234%\"\u003e\n \u003cp\u003e280.62\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"34.28120063191153%\"\u003e\n \u003cp\u003e\u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" width=\"32.859399684044234%\"\u003e\n \u003cp\u003eCr\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"32.859399684044234%\"\u003e\n \u003cp\u003e357.51\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"34.28120063191153%\"\u003e\n \u003cp\u003e\u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" width=\"32.859399684044234%\"\u003e\n \u003cp\u003eFe\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"32.859399684044234%\"\u003e\n \u003cp\u003e374.97\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"34.28120063191153%\"\u003e\n \u003cp\u003e\u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" width=\"32.859399684044234%\"\u003e\n \u003cp\u003eCa\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"32.859399684044234%\"\u003e\n \u003cp\u003e393.88\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"34.28120063191153%\"\u003e\n \u003cp\u003e\u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003c/tbody\u003e\n\u003c/table\u003e\n\u003cp\u003e\u0026nbsp;\u003c/p\u003e\n\u003ctable border=\"1\" cellpadding=\"0\" cellspacing=\"0\" width=\"624\"\u003e\n \u003ctbody\u003e\n \u003ctr\u003e\n \u003ctd colspan=\"3\" valign=\"top\" width=\"100%\"\u003e\n \u003cp\u003e\u003cstrong\u003eTable 3\u0026nbsp;\u003c/strong\u003e\u003cstrong\u003eAverage intensity of trace elements present in T1.\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" width=\"33.333333333333336%\"\u003e\n \u003cp\u003eELEMENTS\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"33.333333333333336%\"\u003e\n \u003cp\u003eMOST INTENSE LINE (nm)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"33.333333333333336%\"\u003e\n \u003cp\u003eAVERAGE INTENSITY (counts)\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" width=\"33.333333333333336%\"\u003e\n \u003cp\u003eAl\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"33.333333333333336%\"\u003e\n \u003cp\u003e279.86\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"33.333333333333336%\"\u003e\n \u003cp\u003e\u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" width=\"33.333333333333336%\"\u003e\n \u003cp\u003eNi\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"33.333333333333336%\"\u003e\n \u003cp\u003e280.62\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"33.333333333333336%\"\u003e\n \u003cp\u003e\u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" width=\"33.333333333333336%\"\u003e\n \u003cp\u003eCr\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"33.333333333333336%\"\u003e\n \u003cp\u003e357.51\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"33.333333333333336%\"\u003e\n \u003cp\u003e\u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" width=\"33.333333333333336%\"\u003e\n \u003cp\u003eFe\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"33.333333333333336%\"\u003e\n \u003cp\u003e374.97\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"33.333333333333336%\"\u003e\n \u003cp\u003e\u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" width=\"33.333333333333336%\"\u003e\n \u003cp\u003eCa\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"33.333333333333336%\"\u003e\n \u003cp\u003e393.88\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"33.333333333333336%\"\u003e\n \u003cp\u003e\u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003c/tbody\u003e\n\u003c/table\u003e\n\u003cp\u003e\u0026nbsp;\u003c/p\u003e\n\u003ctable border=\"1\" cellpadding=\"0\" cellspacing=\"0\" width=\"624\"\u003e\n \u003ctbody\u003e\n \u003ctr\u003e\n \u003ctd colspan=\"3\" valign=\"top\" width=\"100%\"\u003e\n \u003cp\u003e\u003cstrong\u003eTable 4\u0026nbsp;\u003c/strong\u003e\u003cstrong\u003eAverage intensity of trace elements present in T2.\u003c/strong\u003e\u003c/p\u003e\n \u003cp\u003e\u003cstrong\u003e\u0026nbsp;\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" width=\"33.333333333333336%\"\u003e\n \u003cp\u003eELEMENTS\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"33.333333333333336%\"\u003e\n \u003cp\u003eMOST INTENSE LINE (nm)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"33.333333333333336%\"\u003e\n \u003cp\u003eAVERAGE INTENSITY (counts)\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" width=\"33.333333333333336%\"\u003e\n \u003cp\u003eAl\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"33.333333333333336%\"\u003e\n \u003cp\u003e279.86\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"33.333333333333336%\"\u003e\n \u003cp\u003e\u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" width=\"33.333333333333336%\"\u003e\n \u003cp\u003eNi\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"33.333333333333336%\"\u003e\n \u003cp\u003e280.62\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"33.333333333333336%\"\u003e\n \u003cp\u003e\u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" width=\"33.333333333333336%\"\u003e\n \u003cp\u003eCr\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"33.333333333333336%\"\u003e\n \u003cp\u003e357.51\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"33.333333333333336%\"\u003e\n \u003cp\u003e\u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" width=\"33.333333333333336%\"\u003e\n \u003cp\u003eFe\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"33.333333333333336%\"\u003e\n \u003cp\u003e374.97\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"33.333333333333336%\"\u003e\n \u003cp\u003e\u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" width=\"33.333333333333336%\"\u003e\n \u003cp\u003eCa\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"33.333333333333336%\"\u003e\n \u003cp\u003e393.88\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"33.333333333333336%\"\u003e\n \u003cp\u003e\u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003c/tbody\u003e\n\u003c/table\u003e\n\u003cp\u003e\u0026nbsp;\u003c/p\u003e\n\u003ctable border=\"1\" cellpadding=\"0\" cellspacing=\"0\" width=\"624\"\u003e\n \u003ctbody\u003e\n \u003ctr\u003e\n \u003ctd colspan=\"3\" valign=\"top\" width=\"100%\"\u003e\n \u003cp\u003e\u003cstrong\u003eTable 5\u0026nbsp;\u003c/strong\u003e\u003cstrong\u003eAverage intensity of trace elements present in T3.\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" width=\"33.333333333333336%\"\u003e\n \u003cp\u003eELEMENTS\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"33.333333333333336%\"\u003e\n \u003cp\u003eMOST INTENSE LINE (nm)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"33.333333333333336%\"\u003e\n \u003cp\u003eAVERAGE INTENSITY (counts)\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" width=\"33.333333333333336%\"\u003e\n \u003cp\u003eAl\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"33.333333333333336%\"\u003e\n \u003cp\u003e279.86\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"33.333333333333336%\"\u003e\n \u003cp\u003e\u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" width=\"33.333333333333336%\"\u003e\n \u003cp\u003eNi\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"33.333333333333336%\"\u003e\n \u003cp\u003e280.62\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"33.333333333333336%\"\u003e\n \u003cp\u003e\u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" width=\"33.333333333333336%\"\u003e\n \u003cp\u003eCr\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"33.333333333333336%\"\u003e\n \u003cp\u003e357.51\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"33.333333333333336%\"\u003e\n \u003cp\u003e1014\u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" width=\"33.333333333333336%\"\u003e\n \u003cp\u003eFe\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"33.333333333333336%\"\u003e\n \u003cp\u003e374.97\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"33.333333333333336%\"\u003e\n \u003cp\u003e802.85\u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" width=\"33.333333333333336%\"\u003e\n \u003cp\u003eCa\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"33.333333333333336%\"\u003e\n \u003cp\u003e393.88\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"33.333333333333336%\"\u003e\n \u003cp\u003e\u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003c/tbody\u003e\n\u003c/table\u003e\n\u003cp\u003e\u0026nbsp;\u003c/p\u003e\n\u003ctable border=\"1\" cellpadding=\"0\" cellspacing=\"0\" width=\"624\"\u003e\n \u003ctbody\u003e\n \u003ctr\u003e\n \u003ctd colspan=\"3\" valign=\"top\" width=\"100%\"\u003e\n \u003cp\u003e\u003cstrong\u003eTable 6\u0026nbsp;\u003c/strong\u003e\u003cstrong\u003eAverage intensity of trace elements present in T4.\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" width=\"33.333333333333336%\"\u003e\n \u003cp\u003e\u003cstrong\u003eELEMENTS\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"33.333333333333336%\"\u003e\n \u003cp\u003e\u003cstrong\u003eMOST INTENSE LINE (nm)\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"33.333333333333336%\"\u003e\n \u003cp\u003e\u003cstrong\u003eAVERAGE INTENSITY (counts)\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" width=\"33.333333333333336%\"\u003e\n \u003cp\u003eAl\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"33.333333333333336%\"\u003e\n \u003cp\u003e279.86\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"33.333333333333336%\"\u003e\n \u003cp\u003e\u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" width=\"33.333333333333336%\"\u003e\n \u003cp\u003eNi\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"33.333333333333336%\"\u003e\n \u003cp\u003e280.62\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"33.333333333333336%\"\u003e\n \u003cp\u003e\u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" width=\"33.333333333333336%\"\u003e\n \u003cp\u003eCr\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"33.333333333333336%\"\u003e\n \u003cp\u003e357.51\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"33.333333333333336%\"\u003e\n \u003cp\u003e\u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" width=\"33.333333333333336%\"\u003e\n \u003cp\u003eFe\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"33.333333333333336%\"\u003e\n \u003cp\u003e374.97\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"33.333333333333336%\"\u003e\n \u003cp\u003e\u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" width=\"33.333333333333336%\"\u003e\n \u003cp\u003eCa\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"33.333333333333336%\"\u003e\n \u003cp\u003e393.88\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"33.333333333333336%\"\u003e\n \u003cp\u003e\u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003c/tbody\u003e\n\u003c/table\u003e\n\u003cp\u003eThe intensity of Calcium was the highest and Chromium was least in PM1 (i.e., Ca\u0026gt;Al\u0026gt;Ni\u0026gt;Fe\u0026gt;Cr) as shown in Figure 4. The intensity of Calcium was the highest and iron was least in PM2 (i.e., Ca\u0026gt; Cr\u0026gt; Al\u0026gt;Ni\u0026gt;Fe) as shown in Figure 5. The intensity of Calcium was the highest and Chromium was least in T1 (i.e., Ca \u0026gt; Al\u0026gt; Ni\u0026gt; Fe\u0026gt; Cr) as shown in Figure 6. The intensity of Calcium was the highest and Chromium was least in T2 (i.e., Ca \u0026gt; Al\u0026gt; Ni\u0026gt; Fe\u0026gt; Cr) as shown in Figure 7. The intensity of Calcium was the highest and iron was least in T3 (i.e., Ca \u0026gt; Al\u0026gt; Ni\u0026gt;Cr\u0026gt; Fe) as shown in Figure 8. The intensity of Calcium was the highest and iron was least in T4 (i.e., Ca \u0026gt; Al\u0026gt; Ni\u0026gt;Cr\u0026gt; Fe) as shown in Figure 9.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003e\u003cu\u003eComparison of trace elements (Al, Fe, Ca, Ni, and Cr) in commercially packaged tobacco\u003c/u\u003e\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eThe average intensities of the trace elements (Al, Fe, Cr, Ni, and Ca,) in all six groups of \u0026lsquo;paan masala\u0026rsquo; and commercially packaged form of tobacco were compared using the ANOVA Kruskal Wallis test (listed in Table 7). There was statistically significant difference between the concentration of trace elements in the six different samples of \u0026lsquo;paan masala\u0026rsquo; and commercially packaged tobacco (p \u0026lt;0.001). The median and Interquartile range was higher in Ca. We can draw the inference that the concentration of Calcium is highest and least Iron in packaged tobacco, then the level of Aluminium and then the level of Nickel and then Chromium as shown in Fig 10 (Ca\u0026gt;Al\u0026gt;Ni\u0026gt;Cr\u0026gt;Fe).\u003c/p\u003e\n\u003ctable border=\"1\" cellpadding=\"0\" cellspacing=\"0\" width=\"625\"\u003e\n \u003ctbody\u003e\n \u003ctr\u003e\n \u003ctd colspan=\"5\" valign=\"top\" width=\"100%\"\u003e\n \u003cp\u003e\u003cstrong\u003eTable 7\u0026nbsp;\u003c/strong\u003e\u003cstrong\u003eKruskal Wallis test to compare the level of trace elements in commercially packaged tobacco\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" width=\"14.56%\"\u003e\n \u003cp\u003eELEMENTS\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"25.44%\"\u003e\n \u003cp\u003eINTENSITY\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"12%\"\u003e\n \u003cp\u003eMEDIAN\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"23.2%\"\u003e\n \u003cp\u003eINTERQUARTILE RANGE\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"24.8%\"\u003e\n \u003cp\u003eP VALUE\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" width=\"14.56%\"\u003e\n \u003cp\u003eAl\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"25.44%\"\u003e\n \u003cp\u003e\u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"12%\"\u003e\n \u003cp\u003e2552.62\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"23.2%\"\u003e\n \u003cp\u003e4351.17\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"24.8%\"\u003e\n \u003cp\u003e\u0026lt;0.001\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" width=\"14.56%\"\u003e\n \u003cp\u003eNi\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"25.44%\"\u003e\n \u003cp\u003e2877.39 \u0026nbsp;1154.99\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"12%\"\u003e\n \u003cp\u003e1768.37\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"23.2%\"\u003e\n \u003cp\u003e2237.31\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"24.8%\"\u003e\n \u003cp\u003e\u0026lt;0.001\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" width=\"14.56%\"\u003e\n \u003cp\u003eCr\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"25.44%\"\u003e\n \u003cp\u003e\u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"12%\"\u003e\n \u003cp\u003e905.83\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"23.2%\"\u003e\n \u003cp\u003e1931.84\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"24.8%\"\u003e\n \u003cp\u003e\u0026lt;0.001\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" width=\"14.56%\"\u003e\n \u003cp\u003eFe\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"25.44%\"\u003e\n \u003cp\u003e\u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"12%\"\u003e\n \u003cp\u003e923.26\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"23.2%\"\u003e\n \u003cp\u003e717.54\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"24.8%\"\u003e\n \u003cp\u003e\u0026lt;0.001\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" width=\"14.56%\"\u003e\n \u003cp\u003eCa\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"25.44%\"\u003e\n \u003cp\u003e\u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"12%\"\u003e\n \u003cp\u003e39818.45\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"23.2%\"\u003e\n \u003cp\u003e36306.05\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"24.8%\"\u003e\n \u003cp\u003e\u0026lt;0.001\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003c/tbody\u003e\n\u003c/table\u003e"},{"header":"Discussion","content":"\u003cp\u003eOur study aimed to assess elements in commercially packaged forms of tobacco and \u0026lsquo;paan masala\u0026rsquo; that claimed to have no tobacco. To the best of our knowledge Laser-induced breakdown spectroscopy (LIBS) has not been employed in assessing the elemental composition of commercially packaged tobacco and \u0026lsquo;paan masala\u0026rsquo;. Since LIBs is a \u0026ldquo;put and play\u0026rdquo; method, it has been used for various applications such as elemental mapping of trace elements in different hard and soft tissue leading to early diagnosis of malignancies, calculi/ stones, biological or dental material like pellets or powders, and detection of bacteria/ viruses in bioaerosols\u0026nbsp;(12)(13).\u0026nbsp;LIBS aids in the multifaceted detection of elements and does not require sample preparation. Furthermore, it is a noncontact experimental procedure, and all states of matter such as solid, liquid, and gas can be assessed\u0026nbsp;(14).\u003c/p\u003e\n\u003cp\u003eIt is a well-established fact that smokeless tobacco products have a major role to play in causing OPMDs and oral cancer(7). \u003cstrong\u003eChiba, et al\u003c/strong\u003e(15)\u0026nbsp;reviewed various toxic elements in tobacco and smokeless tobacco. They showed the presence of various elements in tobacco products such as Cr, Al, Ni As, Mn, Cu, Pb, Hg, and Zn. Our study showed a similar presence of trace elements that were listed by the authors. The concentration of Aluminium and Nickel were relatively higher and lesser in Chromium.\u0026nbsp;Some trace elements are chemical carcinogens which may be genotoxic or non-genotoxic. Genotoxic carcinogens interact directly with the DNA resulting in chromosomal aberration and DNA damage. While, the latter are chemicals that act directly as tumor promotors, induce inflammatory response, cause immunosuppression, and tissue toxicity. Cr, Be, Ni, Cd, and As are considered carcinogens by International Agency for Research on Cancer (IARC)\u0026nbsp;(8).\u003c/p\u003e\n\u003cp\u003eNickel initiates carcinogenesis by attaching to DNA protein causing mutilated DNA leading to cross-linking of DNA interstrand and DNA protein, and breakage of the DNA strand. Chromium initiates carcinogenesis by producing reactive oxygen species (ROS) and suppressing p53 gene\u0026nbsp;(8)\u0026nbsp;(16).\u003c/p\u003e\n\u003cp\u003eWe also observed highest concentration level of Calcium and lowest concentration of Iron in commercially packaged tobacco and \u0026lsquo;paan masala\u0026rsquo; samples. \u003cstrong\u003eDhaware, et al\u003c/strong\u003e(17)\u0026nbsp;determined the presence of toxic metals such as As, Cd, Cu, and Pb in various types of Indian smokeless tobacco (gutkha, creamy stuff, khaini, mishri, tooth powder, zarda) products using differential pulse anodic stripping voltammetry (DPASV). There were also negligible concentrations of other elements such as Ni, Cr, and Hg. \u003cstrong\u003eMohammed et al\u003c/strong\u003e (18)\u0026nbsp;quantified the presence of various elements in Shisha and Doksha tobacco products using Energy Dispersive X \u0026ndash; ray Fluorescence (EDXRF). They observed higher concentrations of Calcium, and other elements such as Al, Ni, Cr, Fe, Mg, K, Sr, Zn, and Mg. Our study also showed, all the commercially packaged forms of tobacco have a higher concentration of Calcium compared to other elements and presence of carcinogenic elements such as Ni, Cr, and other elements such as Fe, and Al was also observed. Patients having the habit of chewing various smokeless tobacco, and smoking tobacco products with confirmed OSMF were assessed to understand the role of trace elements in the etiopathogenesis of OSMF by \u003cstrong\u003eBagewasi, et al\u003c/strong\u003e(19)\u0026nbsp;and \u003cstrong\u003eKode, et al\u003c/strong\u003e(20). They also stated that there was an elevation of salivary Copper levels in patients having OSMF with the habit of chewing various tobacco products. There was also alteration in Cu/Zn ratio and decreased Fe levels in patients having OSMF with habits.\u003c/p\u003e\n\u003cp\u003eSurprisingly, there was no difference in the elemental composition of commercially packaged tobacco and \u0026lsquo;paan masala\u0026rsquo; packets (that claimed no tobacco) in our study.\u003c/p\u003e"},{"header":"Conclusion","content":"\u003cp\u003eConsumption of smokeless forms of tobacco has increased for the past one decade in India and in southeast Asian regions. Packaged forms of tobacco and betel quid are popularly consumed in India. Our study revealed the presence of various elements such as Calcium, Aluminum, Iron, and known carcinogenic elements such as Nickel and Chromium. Interestingly commercially packaged form (Paan masala), claimed by the manufacturer as tobacco free has similar composition as that of tobacco containing packets. We recommend studying elemental composition of body fluids such as blood, saliva and tissue specimen in patients with deleterious chewing habits. We hope such a study will give us insight into the role of elements in etiopathogenesis of OPMDs and oral cancer.\u003c/p\u003e"},{"header":"Declarations","content":"\u003cp\u003e\u003cstrong\u003e\u003cem\u003eAcknowledgement\u0026nbsp;\u003c/em\u003e\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eThe authors are thankful for the support rendered by the Device Development Program, Department of Science \u0026amp; Technology (DST), Government of India (DST/TDT/DDP-26/2018) and Department of Atomic Energy (DAE), Board of Research in Nuclear Sciences (BRNS), Government of India (34/14/04/2014-BRNS).\u003c/p\u003e"},{"header":"References","content":"\u003col\u003e\n \u003cli\u003e\u003cspan\u003eVini Mary Antony J, Ramani P, Anuja, Sherlin HJ, Jayaraj G, Don, et al. Detection of trace metals among the most commonly available smoked and smokeless tobacco products. J Pharm Sci Res. 2017;9(10):1855\u0026ndash;7.\u003c/span\u003e\u003c/li\u003e\n \u003cli\u003e\u003cspan\u003eGroup IA for R on CW. Personal habits and indoor combustions. Volume\u0026nbsp;100 E, A review of human carcinogens. IARC monographs on the evaluation of carcinogenic risks to humans. 2009;\u003c/span\u003e\u003c/li\u003e\n \u003cli\u003e\u003cspan\u003eWaheed S, Siddique N, Rahman S. Evaluation of trace elements in chewing tobacco and snuff using Instrumental Neutron Activation Analysis (INAA) and Atomic Absorption Spectroscopy (AAS). Radiochim Acta. 2009;97(12):763\u0026ndash;9.\u003c/span\u003e\u003c/li\u003e\n \u003cli\u003e\u003cspan\u003ePatidar KA, Parwani R, Wanjari SP, Patidar AP. Various terminologies associated with areca nut and tobacco chewing: A review. J oral Maxillofac Pathol JOMFP. 2015;19(1):69.\u003c/span\u003e\u003c/li\u003e\n \u003cli\u003e\u003cspan\u003eNiaz K, Maqbool F, Khan F, Bahadar H, Ismail Hassan F, Abdollahi M. Smokeless tobacco (paan and gutkha) consumption, prevalence, and contribution to oral cancer. Epidemiol Health. 2017;39:e2017009.\u003c/span\u003e\u003c/li\u003e\n \u003cli\u003e\u003cspan\u003eIslam S, Muthumala M, Matsuoka H, Uehara O, Kuramitsu Y, Chiba I, et al. How each component of betel quid is involved in oral carcinogenesis: mutual interactions and synergistic effects with other carcinogens\u0026mdash;a review article. Curr Oncol Rep. 2019;21(6):1\u0026ndash;13.\u003c/span\u003e\u003c/li\u003e\n \u003cli\u003e\u003cspan\u003eNiaz K, Maqbool F, Khan F, Bahadar H, Hassan FI, Abdollahi M. Smokeless tobacco (paan and gutkha) consumption, prevalence, and contribution to oral cancer. Epidemiol Health. 2017;39.\u003c/span\u003e\u003c/li\u003e\n \u003cli\u003e\u003cspan\u003eMulware SJ. Trace elements and carcinogenicity: a subject in review. 3 Biotech. 2013;3(2):85\u0026ndash;96.\u003c/span\u003e\u003c/li\u003e\n \u003cli\u003e\u003cspan\u003eAl-rawi NH. Quantitative analysis of trace elements in saliva of oral cancer patients from Iraq. 2015;(January 2005).\u003c/span\u003e\u003c/li\u003e\n \u003cli\u003e\u003cspan\u003eWadhwa SK, Kazi TG, Afridi HI, Talpur FN. Clinica Chimica Acta Interaction between carcinogenic and anti-carcinogenic trace elements in the scalp hair samples of different types of Pakistani female cancer patients. Clin Chim Acta [Internet]. 2015;439:178\u0026ndash;84. Available from: \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttp://dx.doi.org/10.1016/j.cca.2014.10.007\u003c/span\u003e\u003c/span\u003e\u003c/span\u003e\u003c/li\u003e\n \u003cli\u003e\u003cspan\u003eSpeight PM, Khurram SA, Kujan O. Oral potentially malignant disorders: risk of progression to malignancy. Oral Surg Oral Med Oral Pathol Oral Radiol. 2018;125(6):612\u0026ndash;27.\u003c/span\u003e\u003c/li\u003e\n \u003cli\u003e\u003cspan\u003eAnabitarte F, Cobo A, Lopez-Higuera JM. Laser-Induced Breakdown Spectroscopy: Fundamentals, Applications, and Challenges. ISRN Spectrosc. 2012;2012:1\u0026ndash;12.\u003c/span\u003e\u003c/li\u003e\n \u003cli\u003e\u003cspan\u003eGaudiuso R, Melikechi N, Abdel-Salam ZA, Harith MA, Palleschi V, Motto-Ros V, et al. Laser-induced breakdown spectroscopy for human and animal health: A review. Spectrochim Acta Part B At Spectrosc. 2019;152:123\u0026ndash;48.\u003c/span\u003e\u003c/li\u003e\n \u003cli\u003e\u003cspan\u003eYu X, Li Y, Gu X, Bao J, Yang H, Sun L. Laser-induced breakdown spectroscopy application in environmental monitoring of water quality: a review. Environ Monit Assess. 2014;186(12):8969\u0026ndash;80.\u003c/span\u003e\u003c/li\u003e\n \u003cli\u003e\u003cspan\u003eChiba M, Masironi R. Toxic and trace elements in tobacco and tobacco smoke. Bull World Health Organ. 1992;70(2):269\u0026ndash;75.\u003c/span\u003e\u003c/li\u003e\n \u003cli\u003e\u003cspan\u003eRodr\u0026iacute;guez-Tom\u0026agrave;s E, Baiges-Gaya G, Casta\u0026ntilde;\u0026eacute; H, Arenas M, Camps J, Joven J. Trace elements under the spotlight: A powerful nutritional tool in cancer. J Trace Elem Med Biol. 2021;68(March).\u003c/span\u003e\u003c/li\u003e\n \u003cli\u003e\u003cspan\u003eDhaware D, Deshpande A, Khandekar RN, Chowgule R. Determination of toxic metals in Indian smokeless tobacco products. ScientificWorldJournal. 2009;9:1140\u0026ndash;7.\u003c/span\u003e\u003c/li\u003e\n \u003cli\u003e\u003cspan\u003eMohammad AB, Mohammad SHK, Mohammad MK, Khan AS, Al-Hajjaj MS. Quantification of Trace Elements in Different Dokha and Shisha Tobacco Products using EDXRF. J Anal Toxicol. 2019;43(4):E7\u0026ndash;22.\u003c/span\u003e\u003c/li\u003e\n \u003cli\u003e\u003cspan\u003eBagewadi SB, Hirpara DR, Paliwal A, Raiyani BD, Hafiz A, Vasra HD, et al. Estimation of Salivary Copper, Zinc, Iron, and Copper-to-zinc Ratio in Oral Submucous Fibrosis Patients : A Case \u0026ndash; control Study. 2022;2\u0026ndash;5.\u003c/span\u003e\u003c/li\u003e\n \u003cli\u003e\u003cspan\u003eAnkolekar Kode M, Rashmiraj Karjodkar F. Estimation of the serum and the salivary trace elements in OSMF patients. J Clin Diagnostic Res. 2013;7(6):1215\u0026ndash;8.\u003c/span\u003e\u003c/li\u003e\n\u003c/ol\u003e"}],"fulltextSource":"","fullText":"","funders":[],"hasAdminPriorityOnWorkflow":false,"hasManuscriptDocX":true,"hasOptedInToPreprint":true,"hasPassedJournalQc":"","hasAnyPriority":false,"hideJournal":true,"highlight":"","institution":"","isAcceptedByJournal":false,"isAuthorSuppliedPdf":false,"isDeskRejected":"","isHiddenFromSearch":false,"isInQc":false,"isInWorkflow":false,"isPdf":false,"isPdfUpToDate":true,"isWithdrawnOrRetracted":false,"journal":{"display":true,"email":"[email protected]","identity":"researchsquare","isNatureJournal":false,"hasQc":true,"allowDirectSubmit":true,"externalIdentity":"","sideBox":"","snPcode":"","submissionUrl":"/submission","title":"Research Square","twitterHandle":"researchsquare","acdcEnabled":true,"dfaEnabled":false,"editorialSystem":"","reportingPortfolio":"","inReviewEnabled":false,"inReviewRevisionsEnabled":true},"keywords":"oral cancer, smokeless tobacco, trace elements, Laser-Induced breakdown spectroscopy","lastPublishedDoi":"10.21203/rs.3.rs-2695953/v1","lastPublishedDoiUrl":"https://doi.org/10.21203/rs.3.rs-2695953/v1","license":{"name":"CC BY 4.0","url":"https://creativecommons.org/licenses/by/4.0/"},"manuscriptAbstract":"\u003cp\u003e\u003cstrong\u003eBACKGROUND: \u003c/strong\u003eOral cancer is the most common malignancy in many developing countries such as India due to increased consumption of smokeless tobacco. Elemental components in commercially packaged forms of tobacco can play a significant role in the causation of oral cancer.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eAIM: \u003c/strong\u003e\u0026nbsp;Assess trace elements in various types of commercially packaged forms of tobacco using LASER-induced breakdown spectroscopy.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eMATERIALS AND METHODS:\u003c/strong\u003e Two types of ‘Paan masala’ and four types of packaged tobacco were obtained from local ‘paan’ shops. The contents in the packets were made into pellets respectively using a hydraulic press and subjected to elemental analysis using Laser-induced breakdown spectroscopy.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eRESULTS\u003c/strong\u003e: A ten-trial experiment was carried out in all six pellets. The National Institute of Standards and Design (NIST) database was used to assess the emission lines. The elements obtained in both commercially packaged tobacco and ‘paan masala’ were similar. The elements are Calcium, Iron, Aluminium, Nickel, and Chromium.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eCONCLUSION:\u003c/strong\u003eIt is known that substances that cause DNA damage and carcinogenesis, are inorganic elements such as nickel and Chromium. It is clear in our study that these carcinogens are present in the commercially packaged form of tobacco and ‘paan masala’ samples.\u003c/p\u003e","manuscriptTitle":"Assessment of Trace Elements in Commercially Packaged Forms of Tobacco Using Laser-induced Breakdown Spectroscopy","msid":"","msnumber":"","nonDraftVersions":[{"code":1,"date":"2023-03-20 22:08:31","doi":"10.21203/rs.3.rs-2695953/v1","editorialEvents":[{"type":"communityComments","content":0}],"status":"published","journal":{"display":true,"email":"[email protected]","identity":"researchsquare","isNatureJournal":false,"hasQc":true,"allowDirectSubmit":true,"externalIdentity":"","sideBox":"","snPcode":"","submissionUrl":"/submission","title":"Research Square","twitterHandle":"researchsquare","acdcEnabled":true,"dfaEnabled":false,"editorialSystem":"","reportingPortfolio":"","inReviewEnabled":false,"inReviewRevisionsEnabled":true}}],"origin":"","ownerIdentity":"9f5ebad5-dc60-4f4a-8eb8-da6263cb172c","owner":[],"postedDate":"March 20th, 2023","published":true,"recentEditorialEvents":[],"rejectedJournal":[],"revision":"","amendment":"","status":"posted","subjectAreas":[],"tags":[],"updatedAt":"2023-04-03T18:44:09+00:00","versionOfRecord":[],"versionCreatedAt":"2023-03-20 22:08:31","video":"","vorDoi":"","vorDoiUrl":"","workflowStages":[]},"version":"v1","identity":"rs-2695953","journalConfig":"researchsquare"},"__N_SSP":true},"page":"/article/[identity]/[[...version]]","query":{"redirect":"/article/rs-2695953","identity":"rs-2695953","version":["v1"]},"buildId":"7rjqhiLT3MXkJMwkYKINL","isFallback":false,"isExperimentalCompile":false,"dynamicIds":[84888],"gssp":true,"scriptLoader":[]}