The study was carried out to investigate water activity, physicochemical properties and color degree of burley tobacco stored in shelf with different moisture content for 56 days. The study was consist of two experimental designs, that is, moisture content in burley tobacco was kept to 13 to 15 %, 17 to 19 %, and 21 to 23 % under ambient temperature of $20^{\circ}C$ with relative humidity of 24 %, and another group of in burley tobacco was only kept in ambient temperature of $3^{\circ}C$ with relative humidity of 50 % for 56 days with same water content of former treatment. Normal leaves were kept for 56 days under 21 to 23% of moisture content with increased water activity and TSNA(tobacco-specific nitrosamine content. It was considered that favorable leaves conditions are due to favorable microbial growth resulting in increased TSNA content. The total nitrogen content has not changed in two experimental groups for 56 days, otherwise the nitrate form nitrogen and total alkaloid contents were only increased by 21 to 23 % of water content. Degree of pH change was slightly decreased in both storage and water treatment. It was more decreased in 21 to 23% of moisture content than that of other moisture content. In stored leaf tobacco, higher moisture content caused lower brightness, yellow degree, and carbonization quotient.

This study was carried out to investigate the changes of curing condition on microclimate of temperature, relative humidity during curing process of burley tobacco leaves. The developed facility, ridge opening type was designed to open the central top roof. The air-cured variety, (N. tabacum cv KB111) was normally grown at the Eumseong tobacco experimental station in 2007. Mean daily temperature of $3^{\circ}C$ in ridge opening type curing facility was lower than that of conventional, whereas mean daily relative humidity of 12.6 % RH was lower in conventional curing facility for the entire stage of curing. The frequency distribution of optimal air temperature at daytime was higher 37.5 % in ridge opening type curing facility than that of conventional, while that of optimal relative humidity was lower 8.2 %. In the ridge opening type curing facility, the excessive drying leaves were low, however the price per kilogram was high. These results suggest that the new developed curing facility may be applied to improved microclimate inside the curing facility for curing burley.

This study was conducted to evaluate the prediction of sensory property of smoke from leaf chemical property and characterize leaf chemical components for the best tobacco taste's leaves in flue-cured tobacco. For analytical and sensory evaluations, one hundred and forty grades were used. The major leaf chemical components to predict the sensory property of smoke were nicotine for impact, irritation and off taste & odor, and total sugar/nicotine ratio for tobacco taste. Within ${\pm}20%$ range of difference, the predictable probabilities of sensory property of smoke form leaf chemical property were 80.0% for off taste & odor and $91.4{\sim}96.4%$ for impact, irritation and tobacco taste. As a result of K-means cluster analysis on the basis of tobacco taste, the desirable leaf chemical component contents were $2.77{\sim}3.55%$ in nicotine and $5.1{\sim}6.9$ in total sugar/nicotine ratio. This study suggest that the some regression equations may be useful to predict the sensory property of tobacco smoke from a few selected leaf chemical components in flue-cured tobacco and to select the flue-cured tobacco leaves for enhance the tobacco taste of cigarette.

This study was conducted to evaluate the prediction of sensory property of smoke from the leaf chemical property and characterize leaf chemical components for the best tobacco taste's leaves in burley tobacco. For analytical and sensory evaluations, sixteen grades were used. The major leaf chemical components to predict the sensory property of smoke were ether extract for tobacco-like, chloride for impact and total nitrogen/nicotine for irritation. Within ${\pm}20\;%$ range of difference, the predictable probabilities of sensory property of smoke from the leaf chemical properties were 100 % for tobacco-like, impact and irritation. As a result of K-means cluster analysis on the basis of tobacco taste, the desirable leaf chemical component contents were $6.5{\sim}6.8\;%$ in ether extract, $0.25{\sim}0.30\;%$ in chloride and $1.26{\sim}1.54$ in total nitrogen/nicotine ratio. This study suggest that the some regression equations may be useful to predict the sensory components of tobacco smoke from a few selected leaf chemical properties in burley tobacco and to select the burley tobacco leaves for enhance the tobacco taste of cigarette.

This study was conducted to evaluate the prediction of sensory characteristics of smoke from the leaf chemical compounds and characterize leaf chemical components for the best tobacco taste's leaves in oriental tobacco. For analytical and sensory evaluations, seventy two grades were used. Sensory evaluation of tobacco smoke for six attributes were scored on fifteen-point scale by $10{\sim}14$ expert panels trained to estimate smoking quality quantitatively. The major leaf chemical compounds to predict the sensory characteristics of smoke were ether extract for tobacco-like, nicotine for impact and total nitrogen/nicotine ratio for irritation, and total sugar for off taste & odor. Within ${\pm}20%$ range of difference, the predictable probabilities of sensory characteristics of smoke from the leaf chemical compounds were 87.5 % for off taste & odor and $94.4{\sim}98.6\;%$ for tobacco-like, impact and irritation. As a result of K-means cluster analysis on the basis of tobacco taste, the desirable leaf chemical compound contents were $5.9{\sim}8.3\;%$ in ether extract, $1.35{\sim}2.27\;%$ in nicotine and $1.17{\sim}2.24$ in total nitrogen/nicotine ratio. This study suggest that the some regression equations may be useful to predict the sensory characteristics of tobacco smoke with a few selected leaf chemical compounds in oriental tobacco and to select the oriental tobacco leaves by means of enhancing the tobacco taste of cigarette.

This study was conducted to evaluate the effect of additives (antioxidants for free radicals reduction) in cigarette filter treated with various antioxidants (three types of proanthocyanidins and ascorbic acid) and various concentrations of ascorbic acid and loaded with activated carbon on the delivery of free radicals of mainstream smoke (MS) by ESR. Also, we analyzed Hoffmann's analytes andscavenger activity according to the storage time and in vitro cytotoxicity. The analysis of spin number of vapor and particulate phase free radicals in MS are decreased to $14{\sim}24\;%$ and $16{\sim}40\;%$, respectively. As aresult of antioxidant potential for inactivity of vapor and particulate phase free radicals, natural antioxidants were more effective than ascorbic acid. Based on the result of the analysis of Hoffmann's analytes for various antioxidantstreated cigarette filters during the smoking, cigarette filter treated with ascorbic acid showed the lower amount of the deliveries of hydroquinone, isoprene and quinoline in MS than those treated with the other antioxidants. In the significant t-test on the difference of the cytotoxicity among the various antioxidants treated-cigarette filters, there are no significant differences at the 95 % confidence level. Those results indicated that the antioxidants were useful for reducing free radicals in MS because of the fast reaction between antioxidant and free radicals.

This study compares menthol migration from fillers to filters and mainstream smoke in a type of leaf tobacco and according to the moisture content differences at the range of $11{\sim}15\;%$. The leaf tobacco used in this study consisted of Korea flue cured upper leaves B1O (KFUB1), A40R (KFUA4), lower leaves C1L (KFLC1), CD4L (KFLCD4), burley upper leaves A3T (KBUA3), lower leaves D3W (KBLD3), Orient Basma I/III (OB), Orient Izmir BIG (OI), expanded tobacco (KET), and reconstituted sheet (KRC). Menthol migration to the filter and mainstream was measured under constant conditions for 80 days with intervals of 20 days. In the comparison between flue cured types, there were significant differences in the filter parts, as follows. KFUB1(34.4 %) KFUA4(37.4 %), KFLC1(43 %) and KFLCD4 were 55.7 %. In the comparision between other types of leaf tobaccos, KFUB1 was 34.4 % and KET was 52.6 % at filter parts. In the methol transfer to mainstream smoke was $16.5{\sim}24.2\;%$. The menthol migration to filters was measured based on the moisture content of $11{\sim}15\;%$ for the KFUB1 after storing it for 80 days. The menthol migrations were $36{\sim}40\;%$ at the moisture content of $11{\sim}15\;%$, respectively. The transfers to mainstream smoke were $12.8{\sim}15.8\;%$.

The amount of nitrate in the tobacco leaf has been shown to be correlated with the levels of alkaloids and nitrosamines. Also the nitrate content of the tobacco correlated closely with the smoke delivery of nitric oxide and tobacco-specific nitrosamines (TSNAs). These are related with the effect of the reconstituted tobacco leaf(Recon) using the tobacco stems. Adsorption process is gaining interest as one of the effective processes of advanced liquid treatment for liquid containing unnecessary materials. This study is focused on the evaluation of four anion exchangers, a cation exchanger and an activated carbon, as adsorbents for reduction of nitrate anion from Recon extract. In order to analyze the nitrate anion, the IC method used in this work was carried out with a Dionex ICS-2000 system. The effects of dosages of adsorbents and concentration of extract on the removal of nitrate anion were examined. Experimental results showed that for nitrate-anion exchanger, nitrate-cation exchanger and nitrate-activated carbon adsorption system, approximately 70 %, 10 %, and 4 % removal efficiencies were achieved at the Brix 10 and the 20 % addition. Although the activated carbon was little efficient for removal of nitrate ion, the removal of nicotine was very efficient at given conditions.

In vitro toxicity tests such as cytotoxicity, mutagenicity and genotoxicity assay are useful for evaluating the relative toxicity of smoke or smoke condensates obtained from different cigarette configurations. A major disadvantage of these tests is relatively time-consuming, complicated and expensive. Recently, a cell-free glutathione consumption assay (GCA) as a rapid and simple screening method for the toxicity assessment of smoke has been reported by Cahours et al. (CORESTA, 2006). This study was carried out to assess the GCA application capable of predicting the toxicity of gas/vapor phase (GVP) of cigarette smoke and to identify individual compounds responsible for the glutathione (GSH) consumption in smoke. Each GVPs from 2R4F, standard cigarette, carbon filter cigarette (ExC) and new carbon filter cigarette (ExN), test cigarettes were collected by automatic smoking machine and evaluated the relative toxicity by GCA and neutral red uptake (NRU) assay. Toxic compounds existed in smoke were also chosen, relative toxicities of these compounds were screened by using two methods and compared individually. The overall order of toxicity by GCA was 2R4F > ExC > ExN, which was consistent with the result of Neutral Red Uptake assay. The levels of carbonyl compounds of ExN were lower than those of 2R4F and ExC, indicating that GSH consumption was associated with carbonyl compound yields. A major toxicant under current study is acrolein, which contributed to more than half of the GSH consumption. Collectively, the toxicity of GVP determined by GCA method may be mainly attributed to acrolein.

GRLs(Guidance Residue Levels) of agricultural chemicals for tobacco are recommended by the CORESTA Agro-Chemical Advisory Committee guide. In the GRLs list, organochlorine group is one of pesticides commonly used on tobacco cultivation. In this model study, the quantitative correlation in the transfer rate of pesticide residue into tobacco smoke by spiking of organochlorine pesticides to cigarette and pyrolysates were investigated. The spiking concentration referred to the range of GRLs list and the organochlorine pesticides in mainstream smoke were analyzed by GC-MS. For the understanding of the composition variation versus temperature, the behavior of pesticides was investigated by pyrolysis-gas chromatography-mass spectrometry(Py-GC-MS). In this study, the transfer rate of pesticide residue into tobacco smoke at four different spiking concentration and the composition of pyrolysates were analyzed differently. At $10\;{\mu}g/cig$ spiking concentrations, the organochlorine pesticides were transferred into tobacco smoke in $0.02\;{\sim}\;10.19\;%$ each of component and the most of pesticides were pyrolyzed during smoking. It was found that the decomposition compounds from organochlorine pesticides were mainly composed of oxygenated and nitrogenous compounds. This study could estimate that the transfer rate of pesticides into tobacco smoke is very small amount.

Hydrogen cyanide(HCN), formed from pyrolysis of various nitrogenous compounds such as protein, amino acids and nitrate in tobacco, is present in both the particulate phase and vapor phase of cigarette smoke. Typically the determination of HCN in cigarette smoke has been done through colorimetric and electrochemical techniques, such as fluorescence spectrometry, UV-spectrophotometry (UV), continuous flow analyzer (CFA), capillary GC-ECD and ion chromatography (IC). Most of these techniques are known to be time-consuming and some of them lack specificity or sensitivity. The available results from both our laboratory and reported literatures for 2R4F Kentucky reference cigarette, smoked under ISO condition, show a relatively wide variation ranging from 100 to 120 ug/cig of HCN. Especially, the precision and accuracy of the analytical results of HCN tend to get worse in low tar cigarettes and under intense smoking condition. In this paper, a more optimized analytical methods than previous ones are suggested. This method shows lower detection limit and has improved precision and accuracy, so it is applicable for wide tar level cigarettes under intense smoking condition as well as under ISO smoking condition. Important features of this method are improved sample collection and quantification systems such as the number of trapping units, volume, temperature and type of trapping solution. To avoid volatilization loss of HCN in analyzing mainstream smoke, it is highly recommended that pH values of trapping solutions should be maintained over 11 and cold traps should be used in collecting mainstream smoke.

This study was performed to apply the more rapid and accurate sample preparation, and the high selectivity and sensitivity of the analyte detection by gas chromatography-triple quadrupole mass spectrometry (GC-MS/MS). QuEChERS (quick, easy, cheap, effective, rugged and safe) method was validated for 49 agrochemicals in the CORESTA Agrochemical Advisory Committee guide and amenable to GC-MS/MS determination. In QuEChERS method, the effects of sorbents (PSA, $C_{18}$ and GCB) and matrix of the analytes in tobacco types (flue-cured, burley and oriental) were investigated. MS/MS acquisition provided high specificity and selectivity for agrochemicals and low limit of quantification. QuEChERS by using PSA alone and the matrix-matched standards gave good recoveries and RSD values in three types of tobaccos. QuEChERS method was no needed to be complex clean-up procedure and would be used as the fast and easy method for agrochemical residue analysis in tobacco.

Acrylamide has been found in many foods. Acrylamide in foodstuffs were analyzed by a GC/MS after bromination of acrylamide or by a LC/MS for underivatized acylamide. Time consuming and laborious clean up procedures is applied for the purification of the extract, in these methods. In this study, a simple and fast method without clean up step for the analysis of acrylamide in mainstream cigarette smoke was developed by using liquid chromatography-tandem mass spectrometry (LC/MS/MS) and the effects of tobacco leaf constituents on acrylamide content was observed. The analysis of acrylamide in mainstream cigarette smoke started to collect TPM (total particulate matter) from smoking and to extract by 0.1 % acetic acid solution and then to detect by liquid chromatography tandem mass spectrometry using electrospray in the positive mode. The recovery of acrylamide in 2R4F reference cigarette was 98 % and the reproducibility was 2.5 % and the limit of detection was 1.6 ng/mL. Reducing sugars and amino acids are considered to be main precursors of acrylamide in foodstuffs. Cut tobacco contain substantial amounts of reducing sugars and amino acid which may be explained the occurrence of acrylamide in mainstream cigarette smoke. The effects of reducing sugars and total nitrogen studied in an experiment with a various tobacco types. This result indicated that reducing sugars are not limiting factor for acrylamide formation, but the level of acrylamide in cigarette smoke was significantly correlated with the total nitrogen contents.

Gas phase and particulate phase radicals in mainstream cigarette smoke were determined Electron Spin Resonance(ESR) spectroscopy. The free radicals in particulate phase have been investigated by benzene extract of Cambridge Filter Pad containing the smoke condensate. Spin trapping method in conjunction with ESR was used to investigate free radicals in the gas phase of cigarette smoke. Several analytical experiments were conducted in order to determine the optimal conditions for maximum signal intensities and reproducibility of results. All the tests were optimized and normalized using the University of Kentucky 2R4F reference cigarette. The optimal conditions were 0.6 mL for analysis volume of ESR, $4{\sim}5\;mL$ for collection volume of spin-adducts, and PBN for quantification of free radicals in gas phase. The radical levels of Kentucky 2R4F cigarettes were found $2.18{\times}10^{14}\;spins/cig.$ and $2.10{\times}10^{15}\;spins/cig.$ in gas phase.