• Title/Summary/Keyword: Leaf Extract

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Induction of apoptotic cell death in human bladder cancer cells by ethanol extract of Zanthoxylum schinifolium leaf, through ROS-dependent inactivation of the PI3K/Akt signaling pathway

  • Park, Cheol;Choi, Eun Ok;Hwangbo, Hyun;Lee, Hyesook;Jeong, Jin-Woo;Han, Min Ho;Moon, Sung-Kwon;Yun, Seok Joong;Kim, Wun-Jae;Kim, Gi-Young;Hwang, Hye-Jin;Choi, Yung Hyun
    • Nutrition Research and Practice
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    • v.16 no.3
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    • pp.330-343
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    • 2022
  • BACKGROUND/OBJECTIVES: Zanthoxylum schinifolium is traditionally used as a spice for cooking in East Asian countries. This study was undertaken to evaluate the anti-proliferative potential of ethanol extracts of Z. schinifolium leaves (EEZS) against human bladder cancer T24 cells. MATERIALS/METHODS: Subsequent to measuring the cytotoxicity of EEZS, the anti-cancer activity was measured by assessing apoptosis induction, reactive oxygen species (ROS) generation, and mitochondrial membrane potential (MMP). In addition, we determined the underlying mechanism of EEZS-induced apoptosis through various assays, including Western blot analysis. RESULTS: EEZS treatment concentration-dependently inhibited T24 cell survival, which is associated with apoptosis induction. Exposure to EEZS induced the expression of Fas and Fas-ligand, activated caspases, and subsequently resulted to cleavage of poly (ADP-ribose) polymerase. EEZS also enhanced the expression of cytochrome c in the cytoplasm by suppressing MMP, following increase in the ratio of Bax:Bcl-2 expression and truncation of Bid. However, EEZS-mediated growth inhibition and apoptosis were significantly diminished by a pan-caspase inhibitor. Moreover, EEZS inhibited activation of the phosphoinositide 3-kinase (PI3K)/Akt pathway, and the apoptosis-inducing potential of EEZS was promoted in the presence of PI3K/Akt inhibitor. In addition, EEZS enhanced the production of ROS, whereas N-acetyl cysteine (NAC), a ROS scavenger, markedly suppressed growth inhibition and inactivation of the PI3K/Akt signaling pathway induced by EEZS. Furthermore, NAC significantly attenuated the EEZS-induced apoptosis and reduction of cell viability. CONCLUSIONS: Taken together, our results indicate that exposure to EEZS exhibits anti-cancer activity in T24 bladder cancer cells through ROS-dependent induction of apoptosis and inactivation of the PI3K/Akt signaling pathway.

A Mixture of Morus alba and Angelica keiskei Leaf Extracts Improves Muscle Atrophy by Activating the PI3K/Akt/mTOR Signaling Pathway and Inhibiting FoxO3a In Vitro and In Vivo

  • Hyun Hwangbo;Min Yeong Kim;Seon Yeong Ji;Da Hye Kim;Beom Su Park;Seong Un Jeong;Jae Hyun Yoon;Tae Hee Kim;Gi-Young Kim;Yung Hyun Choi
    • Journal of Microbiology and Biotechnology
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    • v.33 no.12
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    • pp.1635-1647
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    • 2023
  • Muscle atrophy, which is defined as a decrease in muscle mass and strength, is caused by an imbalance between the anabolism and catabolism of muscle proteins. Thus, modulating the homeostasis between muscle protein synthesis and degradation represents an efficient treatment approach for this condition. In the present study, the protective effects against muscle atrophy of ethanol extracts of Morus alba L. (MA) and Angelica keiskei Koidz. (AK) leaves and their mixtures (MIX) were evaluated in vitro and in vivo. Our results showed that MIX increased 5-aminoimidazole-4-carboxamide ribonucleotide-induced C2C12 myotube thinning, and enhanced soleus and gastrocnemius muscle thickness compared to each extract alone in dexamethasone-induced muscle atrophy Sprague Dawley rats. In addition, although MA and AK substantially improved grip strength and histological changes for dexamethasone-induced muscle atrophy in vivo, the efficacy was superior in the MIX-treated group. Moreover, MIX further increased the expression levels of myogenic factors (MyoD and myogenin) and decreased the expression levels of E3 ubiquitin ligases (atrogin-1 and muscle-specific RING finger protein-1) in vitro and in vivo compared to the MA- and AK-alone treatment groups. Furthermore, MIX increased the levels of phosphorylated phosphoinositide 3-kinase (PI3K), protein kinase B (Akt), and mammalian target of rapamycin (mTOR) that were reduced by dexamethasone, and downregulated the expression of forkhead box O3 (FoxO3a) induced by dexamethasone. These results suggest that MIX has a protective effect against muscle atrophy by enhancing muscle protein anabolism through the activation of the PI3K/Akt/mTOR signaling pathway and attenuating catabolism through the inhibition of FoxO3a.

Determination of Fomesafen Residue in Agricultural Commodities Using HPLC-UVD/MS (HPLC-UVD/MS를 이용한 농산물 중 fomesafen의 분석)

  • Lee, Su-Jin;Hwang, Young-Sun;Kim, Young-Hak;Nam, Mi-Young;Hong, Seung-Beom;Yun, Won-Kap;Kwon, Chan-Hyeok;Do, Jung-A;Im, Moo-Hyeog;Lee, Young-Deuk;Choung, Myoung-Gun
    • The Korean Journal of Pesticide Science
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    • v.14 no.2
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    • pp.95-103
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    • 2010
  • Fomesafen is a selective herbicide, and used to control annual and perennial broad-leaf grass on soybean and fruit fields in USA and China, but not introduced in Korea yet. So, MRL (Maximum Residue Level), and analytical method of fomesafen were not establishment in Korea. Therefore, this experiment was conducted to establish a determination method for fomesafen residue in crops using HPLC-UVD/MS. Fomesafen residue was extracted with acetone from representative samples of five raw products which comprised hulled rice, soybean, apple, green pepper, and Chinese cabbage. The extract was diluted with saline water, and dichloromethane partition was followed to recover fomesafen from the aqueous phase. Florisil column chromatography was additionally employed for final clean up of the extract. The fomesafen was quantitated by HPLC with UVD, using a Shiseido CAPCELL-PAK UG C18 column. The crops were fortified with fomesafen at 3 levels per crop. Mean recovery ratio were ranged from 87.5% for a 0.4 ppm in hulled rice to 102.5% for a 0.4 ppm in apple. The coefficients of variation were ranged from 0.6% for a 2.0 ppm in hulled rice to 7.7% for a 0.04 ppm in green pepper. Quantitative limit of fomesafen was 0.04 mg/kg in representative 5 crop samples. A LC/MS with selected-ion monitoring was also provided to confirm the suspected residue. Therefore, this analytical method was reproducible and sensitive enough to determine the residue of fomesafen in agricultural commodities.

Effects of Level of Dietary Protein or Carbohydrate on the Economic Characters and Amylase Activity of the Silkworm, Bombyx mori (인공사료의 단백질 및 탄수화물 수준이 가잠의 실용형질과 Amylase활성에 미치는 영향)

  • 문재유
    • Journal of Sericultural and Entomological Science
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    • v.16 no.2
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    • pp.35-53
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    • 1974
  • A series of experiment was conducted to investigate the effects of the level of dietary Protein (soybean meal) and carbohydrate (sucrose) on the growth rate, feed efficiency, nutrients digestibility and amylase activity of larvae at 5th instar. The results obtained from the experimentation are summarized as follow: (1) It was found that the body weight gain, cocoon quality and feed efficiency of the 5th instar larvae were significantly affected by the level of protein and carbohydrate in the artificial diet. Present data revealed that the proper level of dietary protein and carbohydrate would be the most important factor for the optimum growth of larvae. (2) In the amount of diet consumption, the group of larvae gained more body weight consumed more diet than the group gained less amount of body weight. (3) Digestibility of nutrients was improved as the level of protein in the diet increased, although the level of dietary carbohydrate did not exert any positive results. (4) It was observed that the amylase activity in digestive juice was stimulated by the increasement of dietary carbohydrate. However, no correlation between the level of dietary protein and carbohydrate was detected in the amylase activity. (5) Analytical data indicated that the amount of protein or carbohydrate excreted through feces and urine was proportional to the content of protein or carbohydrate in artificial diet. The group o( larvae consumed the diet contained high protein excreted more protein and less nitrogen free extract than did the group received low protein diet. Likewise, the group of larvae fed high dietary carbohydrate excreted more nitrogen free extract and less protein through excreta than the group consumed low carbohydrate diet group. (6) Although the amylase activity of the larvae received the artificial diet was higher than the larvae received natural diet (mulberry leaf), the rate of body weight gam and the quality of cocoon produced from the natural diet group was better than the artificial diet group. It is, thus, concluded that feeding the larvae by the artificial diet may net be recommendable. (7) It was determined that the best level of protein (soybean meal) and carbohydrate (sucrose) in artificial diet was found to be 40g and 0g, respectively when it was fed to the 5th instar larvae. (8) It may be concluded from the results obtained that the artificial diet that could stimulate the activity of amylase nay not be recommendable for the practical larvae feeding Purpose due simply to the Poor economic return from this diet than natural diet.

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Nutritional Components, Texture, and Antioxidant Properties of Lactic Acid Bacteria-Fermented Yakchobugak with Addition of Agro-food Products (유산균을 이용한 발효약초부각의 영양성분, 조직감 및 항산화)

  • Ko, Young-Ran;Shon, Mi-Yae;Wang, Su-Bin;Lee, Kang-Soo;Kang, Seong-Koo;Park, Seok-Kyu
    • Food Science and Preservation
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    • v.16 no.3
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    • pp.405-411
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    • 2009
  • The manufacturing process and quality properties of Lactococcus lactis-fermented yakchobugak (LFY) containing some colored agro-food products were investigated to develop a good organoleptic bugak from the leaf of the medicinal herb. The de-oiling rate of LFY by centrifugation was 37.6%, which was significantly different to the 3.5% value obtained after standing and the 8.9% value obtained with the beating method (p<0.05). Reducing power (RS) increased with addition of increasing levels of ethanol extracts from agro-food powders. The RS of green tea extract-supplemented LFY had the highest value of 0.97 at $500{\mu}g/mL$, and this was significantly different to values obtained using other extracts (p<0.05). At 83.84%, the DPPH (2,2-diphenyl-1-picrylhydrazyl) free-radical scavenging activity of green tea extract-supplemented LFY was the greatest of all samples tested, followed by LFY with woolgeom at 39.48%, LFY with black rice at 28.45%, and LFY with bokbunja at 22.24% all ethanol extracts were added at $50{\mu}g/mL$ (p<0.05). Acid values of green tea and bokbunja LFYs stored in transparent PE bags at $60^{\circ}C$ for 7 days were 1.82% and 2.03%, respectively. Textural hardness values of LFYs were less than 250.62, except for LFYs with black rice and woolgeom, but these values increased $2{\sim}3$-fold after lactic acid fermentation. Carbohydrate and protein content of LFYs were in the ranges $58.95{\sim}64.63%$ and $7.50{\sim}8.68%$, respectively. Lipid and calorie contents of woolgeom LFY had the lowest values of 22.02% and 490 Kcal, respectively.

Ethanol Extract of Mori Folium Inhibits AICAR-induced Muscle Atrophy Through Inactivation of AMPK in C2C12 Myotubes (C2C12 근관세포에서 상엽에 의한 AMPK의 불활성화와 AICAR로 유도된 근위축 억제의 연관성에 관한 연구)

  • Lee, Yu Sung;Kim, Hong Jae;Jeong, Jin-Woo;Han, Min-Ho;Hong, Su Hyun;Choi, Yung Hyun;Park, Cheol
    • Journal of Life Science
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    • v.28 no.4
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    • pp.435-443
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    • 2018
  • AMP-activated protein kinase (AMPK) functions as a metabolic master through regulating and restoring cellular energy balance. In skeletal muscle, AMPK increases myofibril protein degradation through the expression of muscle-specific ubiquitin ligases. Mori Folium, the leaf of Morus alba, is a traditional medicinal herb with various pharmacological functions; however, the effects associated with muscle atrophy have not been fully identified. In this study, we confirmed the effects of AMPK activation by examining the effects of 5-aminoimidazole-4-carboxamide ribonucleotide (AICAR), an activator of AMPK, on the induction of atrophy and expression of atrophy-related genes in C2C12 myotubes. We also investigated the effects of the ethanol extract of Mori Folium (EEMF) on the recovery of AICAR-induced muscle atrophy in C2C12 myotubes. It was found that exposure to AICAR resulted in the stimulation of Forkhead box O3a (FOXO3a); an up-regulation of muscle-specific ubiquitin ligases such as Muscle Atrophy F-box (MAFbx)/atrogin-1 and muscle RING finger-1 (MuRF1), and a down-regulation of muscle-specific transcription factors, such as MyoD and myogenin; with the activation of AMPK. In addition, AICAR without cytotoxicity indicated a decrease in diameter of C2C12 myotubes. However, treatment with EEMF significantly suppressed AICAR-induced muscle atrophy of C2C12 myotubes in a dose-dependent manner as confirmed by a decrease in myotube diameter, which is associated with a reversed stimulation of FOXO3a by the inhibition of AMPK activation. These results indicate that the activation of AMPK by AICAR induces muscle atrophy, and EEMF has preeminent effects on the inhibition of AICAR-induced muscle atrophy through the AMPK signaling pathway.

Anti-oxidant and Anti-inflammatory Properties of Clerodendrum trichotomum Leaf Extracts (누리장나무 잎 추출물의 항산화 및 항염증 활성)

  • Kim, Ji Hye;Song, Hana;Ko, Hee Chul;Lee, Ju Yeop;Jang, Mi Gyeong;Kim, Se Jae
    • Journal of Life Science
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    • v.27 no.6
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    • pp.640-645
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    • 2017
  • Clerodendrum trichotomum (CT) leaves and stems have been used in folk medicine for their anti-hypertension, arthritis, rheumatism, and anti-inflammatory properties. This study was performed to evaluate the potential of CT as an anti-oxidant and anti-inflammatory agent. CT leaves were extracted using 70% ethanol (EtOH). Then, using this extract, a hexane, chloroform ($CHCl_3$), ethyl acetate (EtOAc), and n-butanol (BuOH) fraction was prepared. The polyphenol contents were higher in the EtOAc fraction ($78.08{\mu}g/mg$) and BuOH fraction ($77.54{\mu}g/mg$) compared to the other fractions. Also, these two fractions exhibited strong 1,1-diphenyl-2-picrylhydrazyl (DPPH) and 2,2'-azino-bis-(3-ethylbenzothiazoline-6-sulphonic acid) (ABTS) radical scavenging activities. Xanthine oxidase inhibitory activities were higher in the $CHCl_3$ fraction ($IC_{50}=4.43{\mu}g/ml$) and EtOAc fraction ($IC_{50}=5.69{\mu}g/ml$). Moreover, the EtOAc fraction effectively inhibited nitric oxide (NO) production in lipopolysaccharide (LPS)-stimulated RAW 264.7 cells ($IC_{50}=18.87{\mu}g/ml$). Thus, we investigated the effects of the EtOAc fraction on the expression of pro-inflammatory cytokines, inducible nitric oxide synthase (iNOS), and cyclooxygenase-2 (COX-2) in LPS-stimulated RAW 264.7 cells. The treatment of the EtOAc fraction ($100{\mu}g/ml$) effectively decreased the levels of the tumor necrosis factor ${\alpha}$ ($TNF-{\alpha}$) and interleukin-6 (IL-6), and the expression of inducible nitric oxide synthase (iNOS) and cyclooxygenase-2 (COX-2). These results suggest the potential for CT extract and fractions as promising anti-oxidant and anti-inflammatory agents.

Regeneration of Seedlings under Different Vegetation Types and Effects of Allelopathy on Seedling Establishment of Abies koreana in the Banyabong Peak, Mt. Chiri (지리산(智異山) 구상나무림(林)에서 타감작용(他感作用)이 치수형성(稚樹形成)에 미치는 영향(影響))

  • Kim, Goon Bo;Lee, Kyung Joon;Hyun, Jung Oh
    • Journal of Korean Society of Forest Science
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    • v.87 no.2
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    • pp.230-238
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    • 1998
  • This study was conducted to compare the regeneration of the seedlings under different vegetation types and to identify the presence of allelopathy in Abies koreana(Ak) natural forest in Banyabong Peak(elevation, 1715m), of Mt. Chiri. Twenty quadrats($10m{\times}10m$) were placed in May, 1996 to classify vegetation structure using TWINSPAN. Water-soluble extracts from leaves and soil humus of different vegetation types were collected to test their effects on both seed germination of Ak and mycelial growth of ectomycorrhizal fungi. Phenolic compounds from soil humus were quantified using HPLC. Among the four vegetation types, Sasa borealis(Sb) was found in both Ak-Quercus mongolica(Qm) and Ak-Rhododendron schlippenbachii(Rs) communities. Natural seeding of Ak was $230,000{\pm}90,000seeds/ha$ in 1995 and their germination rate was 25% in an ideal laboratory condition. Density of Ak seedlings less than 5cm in height was 52,000/ha in 1996, while that of seedlings taller than 5cm in height was only 4,000/ha. In the case of Ak-Qm community, density of Ak seedlings with Sb understory was only 7% of the density of seedlings with Rs understary, suggesting the inhibitory effect of Sb. The germination rate of Ak seeds was significantly reduced by leaf extracts of Sb, and Rhododendron mucronulatum var. ciliatum(Rm) and Ak. Soil humus extract of Ak-Qm-Sb subcommunity reduced germination of Ak seeds by 81% and also reduced by 19% the respiration of mycelia of ectomycorrhizal fumgus, Lactarius sp. Among the seven phenolic compounds identified from the soil humus, extract, Ak-Qm-Sb subcommunity contained significantly high content of p-hydroxybenzoic acid, vanillic acid, and syringic acid. Particularly, p-hydroxybenzoic acid was present at 4.2ppm in the Sb roots and at 16.5ppm in the Sb humus, suggesting that it could be the primary allelopathic compound in Abies koreana forests with Sasa borealis understory.

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Control of Cucumber Downy Mildew Using Resistant Cultivars and Organic Materials (저항성 품종과 유기농업자재를 활용한 오이 노균병 방제)

  • Kim, Yong-Ki;Park, So-Hyang;Um, Da-Om;Hong, Sung-Jun;Cho, Jung-Lai;Ahn, Nan-Hee;Shim, Chang-Ki;Kim, Min-Jeong;Park, Jong-Ho;Han, Eun-Jeong;Ko, Byeong-Gu
    • Research in Plant Disease
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    • v.24 no.2
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    • pp.153-161
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    • 2018
  • We selected eight resistant cultivars including 'Heukryongsamcheok', 'Heukgeumsolsamcheok' and 'Gangryeoksamcheok' showing high suppressive effect against cucumber downy mildew (CDM) through plastic film house and field trials in spring and autumn season in 2015. Of them, these three varieties, 'Heukryongsamcheok', 'Heukgeumsolsamcheok' and 'Gangryeoksamcheok' were used to evaluated suppressive effect against CDM by comparing disease severity (area under disease development progress curve, AUDPC) with those of three susceptible varieties under plastic film house condition. AUDPC of three resistant cultivars was in the range of 10.9 to 23.6, meanwhile those of three susceptible cultivars was in the range of 286.7 to 290.3. Consequently, we confirmed that cultivation of the selected resistant cultivars can reduce CDM in vivo. When disease severity (diseased leaf area) was investigated on a resistant cultivar ('Heukryongsamcheok') sprayed without any organic materials and susceptible cultivars sprayed single-, two times- and three times with Bordeaux mixture, sulfur-loess mixture and $Kocide^R$ (copper hydroxide), respectively, it was the lowest in the resistant cultivar plot compared to susceptible cultivar plots sprayed with any organic materials. In addition, we evaluated control effect of the selected resistant cultivar by comparing that of organic materials including lime sulfur mixture, Bordeaux mixture and garlic extract. As a result, Bordeaux mixture showed the highest control effect against CDM. AUDPC of Bordeaux mixture, resistant cultivar, lime sulfur, garlic extract and untreated control was 3.9, 10.6, 95.6, 24.9, and 258.7, respectively. Based on the above-mentioned results, we think that the resistant cultivars and Bordeaux mixture can be effectively used to control CDM as one of control measures under the farmhouse condition.

Changes of Nitrogenous Compound According to the Topping Stage and Harvesting Time in Burley Tobacco(Nicotiana tobacum L.) (버어리종 잎담배 순지르기 시기와 수확시기에 따른 질소화합물의 변화)

  • Jang, Soo-Won;Kim, Jae-Hyun;Park, Chang-Jin;Kim, Yoon-Ha;Lee, In-Jung
    • KOREAN JOURNAL OF CROP SCIENCE
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    • v.56 no.2
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    • pp.146-150
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    • 2011
  • A large amount of nitrogenous compounds are found in burley tobacco and are responsible for peculiar aroma during smoking. Excess in nitrogen or its compounds such as total nitrogen and proteins in burley tobacco gives a strong pungency and make its taste bad. The present study has focused on improving quality of Burley tobacco by regulating topping and harvesting time of leaf to reduce nitrogen compounds in Burley tobacco. In addition, the early-stage of flower topping had lower level of nitrogenous compound, such as total nitrogen and protein nitrogen, and total tobacco-specific nitrosamine (TSNA) contents compared to the button-stage and full flower-stage topping. Also, the contents of nicotine got significantly decreased while the ether extract was increased as the period of topping got delayed. Regarding the brightness among the colors of cured leaves, the color at the full flower-stage topping was the brightest, and there was no particular difference in red and yellow colors. The harvesting amount was decreased as the period of topping got delayed and there was no significant difference between the topping treatments in terms of the exterior quality. As the harvesting time delayed, the total nitrogen, protein nitrogen, nitrate nitrogen, and ammonium nitrogen were decreased while the nicotine contents got increased. Also, the total TSNA contents were decreased as the harvesting time delayed. However, ether extract was increased. Among the colors of cured leaves, the brightness and degree of yellow color were significantly increased as the harvesting time delayed, and there was no particular difference in the degree of red color. The harvesting amount was decreased along with the delayed harvesting time; however, the harvest of the matured and mellow leaves was higher in terms of price, compared to that of the immature leaves. For the reduction of nitrogenous compound and TSNA contents in Burley tobacco, the topping was examined to be more effective in terms of improvement in chemical contents and quality regardless of the decreased harvesting amount.