This study was carried out to investigate the changes of essential oils in flue-cured leaf tobacco during aging for 21 months. The threshed leaf tobacco(BlO) produced in 2002 crop year was aged for 21 months in the warehouse of Oc-Cheon Leaf Tobacco Processing Factory. The leaf tobacco were sampled at three month intervals for analysis of volatile compounds. Volatile compounds were identified by GC/MS and comparison of gas chromatographic retention time with those of the authentic standard. The total of 75 compounds from the steam volatile concentrate of the flue-cured leaf tobacco were identified; they were 15 hydrocarbons, 12 alcohols, 3 aldehydes, 18 ketones, 7 esters, 10 acids, 3 phenols, 4 furans, 2 pyrrols and 1 pyridine. The major components of essential oil were neophytadiene, solanone, megastigmatrienone and phytol. After a aging period of 21 months, most of volatile compounds showed a gradual increasing tendency.

This study was carried out to investigate the changes of free sugars, non-volatile organic and fatty acids in flue-cured leaf tobacco during aging. The threshed leaf tobacco(B1O and C1L) produced in 2002 crop year was aged for 21 month in warehouse of Oc-Cheon Leaf Tobacco Processing Factory. The leaf tobacco were sampled at three month intervals for analysis of free sugars, non-volatile organic and fatty acids. The major free sugars of flue-cured were glucose and fructose regardless of tobacco grades. After aging period of 21 months, the contents of glucose and fructose showed a tendency to slightly decrease, and there was no appreciable change in the contents of sucrose in B1O and C1L grades. The major compound of non-volatile organic acid and fatty acid were malic, oxalic and citric, linolenic, linoleic and palmitic acid regardless of tobacco grade. After aging period of 21 months, the contents of malic, malonic, oxalic and fumaric acid showed a tendency to slightly decrease, whereas succinic acid was slightly increased, and maleic acid was not changed in B1O and C1L grades. The decreases in fatty acid contents in B1O and C1L grade tobacco loaves after 21 month aging were 16.5% and 9.8%, respectively. The decreases in linolenic acid contents in two grades were the highest, showing 22.1% and 12.0% reduction after 21 month aging.

The aim of this study is to develop a simple and rapid field method for the determination of chloride content in tobacco green or cured leaves. To set up a practical process, several factors such as the proper sampling position of each leaf, the size of leaf lamina pieces for blending, the substitution of deionized water fur the extracting solution, 0.1N $HNO_3$, the submerging durations before blending, and the standing times before measuring were studied using flue-cured and burley tobacco with a stable and reproducible Chloride Meter, KRK CL-2Z. From the results, chloride contents of tobacco leaves could be analyzed with the Chloride Meter(CL-2Z) using the suspension of 14 mm diameter leaf disks after soaking in distilled water for 30 min., blending with a mini-blender for 30 seconds. The chloride contents measured in this way showed highly significant correlations with ones by the conventional potentiometric method using an ion meter-chloride electrode and $AgNO_3$ titration. In our method, the procedures such as drying, powdering and weighing samples, and using chemical reagents for extraction (0.1N $HNO_3$) and titration(0.028N $AgNO_3$) could be eliminated. These results suggest that the newly constructed method using the Chloride Meter is more efficient to determine the chloride content of tobacco leave in comparison with the one by the Ion $Meter:AgNO_3$ titration.

AFLP(Amplified Fragment Length Polymorphism) analysis was conducted to cultivars of tobacco, Nicotiana tabacum in order to select the cultivar-specific markers. AFLP results using 12 primer sets revealed genetic diversity among 12 field grown tobacco cultivars. Polymorphic fragments amplified by PCR was purified and cloned to identify their nucleotide sequences. From the sequences of them, 40 primer sets were designed to select cultivar-specific markers. When genomic DNA isolated from tobacco were used as PCR template, a set of primers, BrSF/BrSR showed Burley-specific band patterns. The results indicate that AFLP technique used in this experiments is useful for identifying tobacco cultivars in a rapid manner.

A mutant of Potato vims Y (PVY) was occurred in PVY resistance flue-cured tobacco breeding line KF0402 $(TC1146{\times}KF117)$ showing vein necrosis at Suwon in Korea. This isolate, PVY-SWM, was differentiated from other PVY based on biological properties and nucleotide sequence analyses of coat protein gene. PVY-SWM caused typical symptoms on 21 indicator plants as compared to the PVY-TOJC37. Remarkably, the PVY-SWM induced distinctly different symptom of systemic vein necrosis on tobacco cultivars V.SCR, PBD6, TN86, TN90, Virgin A Mutant (VAM), Wislica, NC744, KB108 and KB111, which were reported to have the recessive potyvirus resistance gene va. In RT-PCR assays with specific primers for detection of PVY, a single band of about 800bp in length was produced. The amplified DNA was cloned and the nucleotide sequence was determined. The coat protein gene of PVY-SWM showed 88.4%-99.0% and 92.5%-98.5% identities to the 12 different PVY isolates of Genbank Database at the nucleotide and amino acidi respectively. Multiple alignments as well as cluster dendrograms of PVY-SWM isolate revealed close phylogenetic relationship to the $PVY^{NTN}$ subgroup.

It is very important to add uniformly casing materials on tobacco for taste and flavor. However, analysis of casing materials was spent much time, effort and money. The object of this study was the development of a rapid method for the determination of glycerine, propylene glycol(PG), sucrose, glucose, fructose and water in the casing materials using the NIR transmittance method. Hundreds of calibration samples, with extended ranges (50%, 75%, 100%, 125%, and 150% of standard addition) in each constituent, were prepared in the casing materials at the various temperatures $(25^{\circ}C\;and\;30^{\circ}C)$. Calibration equation was developed by modified partial least square (MPLS) method using second derivative. The standard error of calibration and $R^2$ between added value and NIR estimated value results were $0.007{\sim}0.034\;and\;0.996{\sim}1.000$ for the casing sample set, respectively. The standard error of prediction and R2 between added value and NIR estimated value results were $0.010{\sim}0.034\;and\;0.997{\sim}1.000$ for the casing sample set, respectively. The analysis result was not different significantly between the NIR and added value. These results show that the NIR measurement system is an effective tool to ensure quality on the casing materials.

The pulping process in a papermaking process for a reconstituted tobacco sheet (RECON) was evaluated. For the practical information, the experiments were conducted in a real mill. Two pulping conditions were chosen and changed depending on the feasibility, such as the pulping temperature and the properties of pulping water. The higher pulping temperature resulted in the easier dissolution of solubles from the raw materials. However, the efficiency of screw press process was decreased by the higher pulping temperature, which resulted in the reduction of the HWS and the concentration in the #1 screw press filtrate. The addition of SEL(Strong extracted liquor) to pulper affected the efficiency of dissolution and the final concentration of #1 screw press filtrate. Although the higher SEL resulted in the less dissolution of the solubles from the raw materials, the concentration of the #1 screw press filtrate was greatly increased by the higher addition of SEL, which could provide much better efficiency to the following evaporation process of the filtrate and could increase the whole productivity.

The grades of burley tobacco stems and scraps were classified followed by the nicotine and nitrate contents by using the cluster analysis. The chemical components of the burley tobacco stems and scraps gathered from 2002 to 2005 were analyzed with auto analyzer. According to the nicotine contents and the nitrate contents, the burley stems and scraps could be classified three groups, respectively. In case of the burley scraps, the AB3T, AB4TR, B1T and B2T grades belonged to the $1^{st}$ a group. The C1W and C2W grades belonged to the $2^{nd}$ group and the CD3W and CD4TR belonged to the $3^{rd}$. In case of the burley stems, the AB3T and AB4TR grades belonged to the $1^{st}$ group. The B1T, B2T, C1W and C2W grades belonged to the $2^{nd}$ group and the CD3W and CD4TR belonged to the $3^{rd}$ group. This classification of raw materials depending on the similarity in the chemical components might be helpful to control the properties of the Reconstituted Tobacco sheet.

This study was carried out to improve the analytical methods for determination of organic acids in tobacco leaf by HPLC. The samples for HPLC analysis were pre-treated by means of SPE. The calibration curve for each acid was linear and $R^2$ values ranged from 0.9990 to 1.0000. The limit of detection were obtained from the calibration curves and their values were between 1.39 to $4.87{\mu}g/mL$. Recovery rates of organic acids were between 88.6 % to 98.5 %. The concentrations of organic acids among the various tobacco leaves were compared to the concentration of organic acids, were in the order oriental, burley, flue-cured tobacco. In the case of flue-cured and oriental tobacco leaves, the order of concentration of organic acids was malic acid, oxalic acid, citric acid. But in the case of burley tobacco leaves, the order of concentration of organic acids was citric acid, oxalic acid, malic acid.

This study was conducted to determine the amount of unprotonated nicotine in cigarette mainstream smoke and to investigate its relationship to level of filter ventilation and machine smoking conditions. Unprotonated nicotine from TPM trapped on a Cambridge filter pad(CFP) was adsorbed by Carboxen/PDMS SPME fiber, thermally desorbed and determined by GC/MS. 2R4F reference cigarette, twelve commercial brands from the Korean market and five test cigarette samples, which had the same tobacco blend with different levels of filter ventilation, were analyzed for unprotonated nicotine. In commercial brands, the amount of unprotonated nicotine changed slightly depending on the pH values of smoke, and decreased as the tar level increased. filter ventilation in these commercial cigarettes was $28{\sim}80%$ and the higher filter ventilation increased relative unprotonated nicotine levels, but not significantly. However, in five test cigarettes with different filter ventilation$(0{\sim}70%)$, unprotonated nicotine levels increased almost linearly with the level of filter ventilation. Concentrations of unprotonated nicotine in mainstream smoke generally increased in order $HC\;<\;ISO\;{\leq}\;MDPH$ machine smoking conditions. The ratio of unprotonated nicotine to total nicotine among $cigarettes({\alpha}_{fb})$ increased in order RC < MDPB < ISO conditions. Concentrations of unprotonated nicotine varied with three machine smoking conditions.

A measurement uncertainty for analysis of volatile organic compound (benzene) in cigarette mainstream smoke was carried out. In this study one point re-calibration method was used to estimate uncertainty for benzene. The measurement uncertainty was calculated based on the uncertainty sources of each analysis step, quality appraisal sources, drift and repeatability. As a result, the concentration and expanded uncertainty of benzene in cigarette mainstream smoke were measured as $38.08{\pm}4.36{\mu}g/cig$. Relative uncertainty of drift and repeatability obtained were 5% and 3%, respectively.

A new procedure has been developed for the quantitation of aromatic amines in mainstream cigarette smoke. Two solid-phase extraction (SPE) clean up steps, using a different retention mechanisms, are required to process the samples. The first step used a cation-exchange cartridge, followed by a second step that used a cartridge with a hydrophobic retention character. The aromatic amines eluted from the second SPE cartridge are derivatized with pentafluoropropionic anhydride. This new method have advantages over other reported techniques, being sensitive, robust, and easily automated. The detection limits were ranged from 0.12 ng/mL for 1-aminonaphthalene to 0.16 ng/mL for 3-aminobiphenyl and the recoveries were from 97 to 106%. Compared with other reports for analysis of 2R4F reference cigarette, this method shows a close analytical data and good repeatability.

Main tobacco diseases were surveyed on major flue-cured leaf tobacco fields throughout South Korea in 2006. Mosaic caused by potato virus Y and bacterial wilt caused by Ralstonia solanacearum were most severe during harvest season. During last ten years, the damage by tobacco mosaic virus was reduced but the incidence of bacterial wilt increased. These changes of the disease incidences coincide with release time of the tobacco cultivar resistant to the tobacco mosaic virus but susceptible to bacterial wilt pathogen. Wild fire(Pseudomonas syringae pv. tabaci strain ungulate) occurred severely at Kangwon province, though the symptom of wild fire with yellow halo was not observed.