Analysis of coumarin and angelica lactones in smokeless tobacco products.
McAdam Kevin,Enos Trevor,Goss Carol,Kimpton Harriet,Faizi Arif,Edwards Steve,Wright Christopher,Porter Andrew,Rodu Brad
Chemistry Central journal
Differences in health risks between different styles of smokeless tobacco products (STPs) have prompted interest in their relative levels of toxic chemicals. We report here the development of methods for the analysis of STPs for coumarin and for α-angelica lactone (α-AL), both of which have been included in various published lists of tobacco toxicants. We have also determined the concentrations of these lactones in commercial STPs from the US and Sweden, representing 80-90% of the 2010 market share for all the major STP categories in these two countries: 65 products (plus two reference products) for coumarin and 66 commercial products for α-AL. For coumarin, methanol extracts of the STPs were analysed by HPLC/MS/MS. The lower limit of quantification (LOQ) and limit of detection (LOD) were, respectively, 100 and 30 ng coumarin/g of STP on a wet weight basis (WWB). Alpha-AL was determined via direct headspace GC/MS. The LOQ and LOD were 65 and 30 ng/g WWB respectively. Coumarin was detected In 3/33 Swedish snus, 5/13 US chewing tobaccos, 16/16 moist snuffs and 5/6 dry snuffs. Concentrations in those samples with quantifiable coumarin contents ranged from 186 to 1656 ng/g WWB. Concentrations of coumarin measured in this study were consistent with levels naturally found in tobacco. None of the STPs analysed would significantly contribute to coumarin exposure in consumers compared with dietary sources, and estimated exposure levels were 1000× lower than the European Food Safety Authority Tolerable Daily Intake. Hence the relevance of coumarin to the toxicity of STPs and its inclusion in the FDA's list of harmful and potentially harmful compounds list is questionable. Measurements of α-AL in these STPs found that the majority did not have quantifiable contents, however, for three STPs concentrations of α-AL were above the LOQ (116-140 ng/g WWB) and for four other STPs concentrations of α-AL could be estimated between the LOD and LOQ. Beta-angelica lactone was tentatively identified in three of the STPs but the levels could not be reliably quantified. The levels of α-AL in tobacco products are reported here for the first time, but the relevance of α-AL to the toxicity of STPs is also highly questionable given that it has GRAS status as a permitted food additive.
Tobacco-specific nitrosamines: A literature review.
Konstantinou Evangelia,Fotopoulou Foteini,Drosos Athanasios,Dimakopoulou Nektaria,Zagoriti Zoi,Niarchos Athanasios,Makrynioti Dimitra,Kouretas Dimitrios,Farsalinos Konstantinos,Lagoumintzis George,Poulas Konstantinos
Food and chemical toxicology : an international journal published for the British Industrial Biological Research Association
Cigarette smoke is a complex mixture of chemicals, including several tobacco-specific nitrosamines (TSNA). Most TSNA are formed in tobacco during the post-harvest period, while a number are produced when a cigarette is burned. Considerable evidence supports the role of TSNA important causative factors for cancers of the lung, pancreas, esophagus, and oral cavity in people who use tobacco products. Of the known TSNA, nicotine-derived nitrosamine ketone (NNK) and N-nitrosonornicotine (NNN) are the most carcinogenic. Other TSNA include N'-nitrosoanatabine (NAT) and N-nitrosoanabasine (NAB). New tobacco products (e.g., e-cigarettes) designed to attract consumers who are concerned about the health effects of tobacco have been appearing on the market. Several studies have reported that certain TSNA have been detected in the replacement liquids and vapour of e-cigarettes, but the levels are generally considerably lower than in tobacco cigarettes. Additionally, the FDA recently announced its intention to regulate TSNA in e-cigarettes, cigar tobacco and pipe tobacco. With the rise of new technologies for reducing the use of tobacco products-such as e-cigarettes- to evaluate exposure levels to these harmful chemicals over time, researchers will be monitoring levels of TSNA in the body as a result of the use of these devices.
Chemical and toxicological characteristics of conventional and low-TSNA moist snuff tobacco products.
Song Min-Ae,Marian Catalin,Brasky Theodore M,Reisinger Sarah,Djordjevic Mirjana,Shields Peter G
Use of smokeless tobacco products (STPs) is associated with oral cavity cancer and other health risks. Comprehensive analysis for chemical composition and toxicity is needed to compare conventional and newer STPs with lower tobacco-specific nitrosamines (TSNAs) yields. Seven conventional and 12 low-TSNA moist snuff products purchased in the U.S., Sweden, and South Africa were analyzed for 18 chemical constituents (International Agency for Research on Cancer classified carcinogens), pH, nicotine, and free nicotine. Chemicals were compared in each product using Wilcoxon rank-sum test and principle component analysis (PCA). Conventional compared to low-TSNA moist snuff products had higher ammonia, benzo[a]pyrene, cadmium, nickel, nicotine, nitrate, and TSNAs and had lower arsenic in dry weight content and per mg nicotine. Lead and chromium were significantly higher in low-TSNA moist snuff products. PCA showed a clear difference for constituents between conventional and low-TSNA moist snuff products. Differences among products were reduced when considered on a per mg nicotine basis. As one way to contextualize differences in constituent levels, probabilistic lifetime cancer risk was estimated for chemicals included in The University of California's carcinogenic potency database (CPDB). Estimated probabilistic cancer risks were 3.77-fold or 3-fold higher in conventional compared to low-TSNA moist snuff products under dry weight or under per mg nicotine content, respectively. In vitro testing for the STPs indicated low level toxicity and no substantial differences. The comprehensive chemical characterization of both conventional and low-TSNA moist snuff products from this study provides a broader assessment of understanding differences in carcinogenic potential of the products. In addition, the high levels and probabilistic cancer risk estimates for certain chemical constituents of smokeless tobacco products will further inform regulatory decision makers and aid them in their efforts to reduce carcinogen exposure in smokeless tobacco products.
Comprehensive chemical characterization of Rapé tobacco products: Nicotine, un-ionized nicotine, tobacco-specific N'-nitrosamines, polycyclic aromatic hydrocarbons, and flavor constituents.
Stanfill Stephen B,Oliveira da Silva André Luiz,Lisko Joseph G,Lawler Tameka S,Kuklenyik Peter,Tyx Robert E,Peuchen Elizabeth H,Richter Patricia,Watson Clifford H
Food and chemical toxicology : an international journal published for the British Industrial Biological Research Association
Rapé, a diverse group of smokeless tobacco products indigenous to South America, is generally used as a nasal snuff and contains substantial amount of plant material with or without tobacco. Previously uncharacterized, rapé contains addictive and harmful chemicals that may have public health implications for users. Here we report % moisture, pH, and the levels of total nicotine, un-ionized nicotine, flavor-related compounds, tobacco-specific N-nitrosamines (TSNAs) and polycyclic aromatic hydrocarbons (PAHs) for manufactured and hand-made rapé. Most rapé products were mildly acidic (pH 5.17-6.23) with total nicotine ranging from 6.32 to 47.6 milligram per gram of sample (mg/g). Calculated un-ionized nicotine ranged from 0.03 to 18.5 mg/g with the highest values associated with hand-made rapés (pH 9.75-10.2), which contain alkaline ashes. In tobacco-containing rapés, minor alkaloid levels and Fourier transform infrared spectra were used to confirm the presence of Nicotiana rustica, a high nicotine tobacco species. There was a wide concentration range of TSNAs and PAHs among the rapés analyzed. Several TSNAs and PAHs identified in the products are known or probable carcinogens according to the International Agency for Research on Cancer. Milligram quantities of some non-tobacco constituents, such as camphor, coumarin, and eugenol, warrant additional evaluation.
Chemical properties investigation of commercial cigarettes by a "pseudo" targeted method using GC-MS-selected ions monitoring.
Li Yong,Pang Tao,Li Yanli,Ye Guozhu,Lu Xin,Xu Guowang
Journal of separation science
A "pseudo" targeted method using GC-MS-selected ions monitoring was applied to investigate the chemical characteristics of commercial cigarettes made in China and foreign countries. To identify the components and define the quantative ions for SIM acquisition, a quality control sample was analyzed using GC-MS full scan. Acquired data were treated with a homemade software. A peak table with 312 components and their related quantitation ions was achieved for SIM acquisition. Structure elucidation was performed using library searching, retention index, standard compounds, and fitted retention time. The fitted retention time was calculated by a linear correction curve obtained using measured and library retention time to verify compounds. A total of 90 compounds were elucidated. Chemical characteristics of different cigarette brands were investigated. The data acquisition was carried out in SIM mode. The principal component and the hierarchical clustering analyses showed that the Chinese domestic flue-cured cigarettes were significantly different from the domestic blended, the foreign flue-cured, and blended cigarettes. Sixty-seven differential compounds were defined using the nonparametric Mann-Whitney test and the group blending samples comparison. Chinese domestic flue-cured cigarettes have higher concentration of saccharides and lower concentration of organic acids and amino acids.
Headspace solid-phase microextraction coupled to comprehensive two-dimensional gas chromatography-time-of-flight mass spectrometry for the analysis of aerosol from tobacco heating product.
Savareear Benjamin,Lizak Radoslaw,Brokl Michał,Wright Chris,Liu Chuan,Focant Jean-Francois
Journal of chromatography. A
A method involving headspace solid-phase microextraction (HS-SPME) and comprehensive two-dimensional gas chromatography coupled to time-of-flight mass spectrometry (GC×GC-TOFMS) was developed and optimised to elucidate the volatile composition of the particulate phase fraction of aerosol produced by tobacco heating products (THPs). Three SPME fiber types were studied in terms of extraction capacity and precision measurements. Divinylbenzene polydimethylsiloxane appeared as the most efficient coating for these measurements. A central composite design of experiment was utilised for the optimization of the extraction conditions. Qualitative and semi-quantitative analysis of the headspace above THP aerosol condensate was carried out using optimised extraction conditions. Semi-quantitative analyses of detected constituents were performed by assuming that their relative response factors to the closest internal standard (t) were equal to 1. Using deconvoluted mass spectral data (library similarity and reverse match >750) and linear retention indices (match window of ±15 index units), 205 peaks were assigned to individual compounds, 82 of which (including 43 substances previously reported to be present in tobacco) have not been reported previously in tobacco aerosol. The major volatile fraction of the headspace contained ketones, alcohols, aldehydes, alicyclic hydrocarbons alkenes, and alkanes. The method was further applied to compare the volatiles from the particulate phase of aerosol composition of THP with that of reference cigarette smoke and showed that the THP produced a less complex chemical mixture. This new method showed good efficiency and precision for the peak areas and peak numbers from the volatile fraction of aerosol particulate phase for both THP and reference cigarettes.
Effects of enzymatic browning reaction on the usability of tobacco leaves and identification of components of reaction products.
Chen Yanjie,Zhou Junfei,Ren Ke,Zou Congming,Liu Junjun,Yao Guangmin,He Jianshen,Zhao Gaokun,Huang Wei,Hu Binbin,Chen Yi,Xiong Kaisheng,Jin Yan
The enzyme browning reaction results in grey speckles on tobacco leaves, which impairs the value and industrial usability of tobacco leaves. To demonstrate the influences of different browning degrees (BDs) of tobacco leaves on the usability of different cultivars and positions and identified structure of brown (grey) matter, we selected three flue-cured tobacco cultivars (K326, Yunyan87, and Honghuadajinyuan (Hongda)) and set four different BDs (<25%, 25% to 50%, 50% to 75%, and >75%). Indices related to: economic traits, chemical components, physical properties, and sensory quality of tobacco leaves with different cultivars were evaluated. Moreover, by utilising thin-layer chromatography and high-performance liquid chromatography, we analysed and identified the structure of the grey matter in terms of chemical composition. The experimental results show that the main component of grey speckles on tobacco leaves is 3-acetyl-6,7-dimethoxycoumarin (YC-ZJF). With the increase of BD, the amount of total sugar and reducing sugar, output value, the proportion of superior tobacco, shatter resistance index, and sensory evaluation score of the three cultivars significantly decrease, while the starch content increases significantly. The changes in protein, total nitrogen, and nicotine are insignificant with changing BD. In addition, other indices show different trends for different cultivars of flue-cured tobacco. After separation and identification of the components of grey speckled leaves, it is proved that the substance derived from grey speckles on tobacco leaves is YC-ZJF. The research is important to the study of browning mechanisms in tobacco leaves and provides corresponding targets for strategies to reduce browning thereof.