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Effect of Nardostachyos Rhizoma on Apoptosis, Differentiation and Proliferation in HL-60 cells

  • Ju Sung-Min;Lee Jun;Choi Ho-Seung;Yoon Sang-Hak;Kim Sung-Hoon;Jeon Byung-Hun
    • Journal of Physiology & Pathology in Korean Medicine
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    • v.20 no.1
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    • pp.163-170
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    • 2006
  • Nardostachyos Rhizoma (N. Rhizoma) belonging to the family Valerianaceae has been anti-arrhythmic effect, and sedation to the central nerve and a smooth muscle. We reported that the water extract of N. Rhizoma induced apoptotic cell death and differentiation in human promyelocytic leukemia (HL-60) cells. Cytotoxicity of N. Rhizoma was detected only in HL-60 cells (IC50 is about 200 ${\mu}g/ml$). The cytotoxic activity of N. Rhizoma in HL-60 cells was increased in a dose-dependent manner. We used several measures of apoptosis to determine whether these processes were involved in N. Rhizoma-induced apoptotic cell death. The high-dose (200 ${\mu}g/ml$) treatment of N. Rhizoma to HL-60 cells showed cell shrinkage, cell membrane blobbing, apoptotic bodies, and the fragmentation of DNA, suggesting that these cells underwent apoptosis. Treatment of HL-60 cells with N. Rhizoma time-dependently induced activation of caspase-3, caspase-8, and caspase-9 and proteolytic cleavage of poly(ADP-ribose) polymerase. Also, we investigated the effect of N. Rhizoma on cellular differentiation and proliferation in HL-60 cells. Differentiation and proliferation of HL-60 cells was determined through expression of CD11b and CD14 surface antigens using flow cytometry and nitroblue tetrazolium (NBT) assay, and through analysis of cell cycle using propidium iodide assay, respectively. N. Rhizoma induced the differentiation of HL-60 at the low-dose (100 ${\mu}g/ml$) treatment, as shown by increased expression of differentiation surface antigen CD11b, but not CDl4 and increased reducing activity of NBT. When HL-60 cells were treated with N. Rhizoma at concentration of $50{\mu}g/ml\;and\;100{\mu}g/ml$, NBT-reducing activities induced approximately 1.5-fold and 20.0-fold as compared with the control. In contrast, HL-60 cells treated with the N. Rhizoma-ATRA combination showed markedly elevated levels of 26.3-fold at $50{\mu}g/ml$ N. Rhizoma-0.1 ${\mu}M$ ATRA combination and 27.5-fold at 50 ${\mu}g/ml$ N. Rhizoma-0.2 ${\mu}M$ ATRA combination than when treated with N. Rhizoma alone or ATRA alone. It may be that N. Rhizoma plays important roles in synergy with ATRA during differentiation of HL-60 cells. DNA flow-cytometry indicated that N. Rhizoma markedly induced a G1 phase arrest of HL-60 cells. N. Rhizoma-treated HL-60 cells increased the cell population in G1 phase from 32.71% to 42.26%, whereas cell population in G2/M and S phases decreased from 23.61% to 10.33% and from 37.78% to 33.98%, respectively. We examined the change in the $p21^{WAF1/Cip1}\;and\;p27^{Kip1}$ proteins, which are the CKIs related with the G1 phase arrest. The expression of the CDK inhibitor $p27^{Kip1},\;but\;not\;p21^{WAF1/Cip1}$ were markedly increased by N. Rhizoma. Taken together, these results demonstrated that N. Rhizoma induces apoptotic cell death through activation of caspase-3, and potently inhibits the proliferation of HL-60 cells via the G1 phase cell cycle arrest in association with $p27^{Kip1}$ and granulocytic differentiation induction .

Analysis of Clonorchis sinensis antigens and diagnosis of clonorchiasis using monoclonal antibodies (단세포군 항체를 이용한 간흡충 항원의 분석 및 간흡충증의 진단)

  • Yong, Tae-Sun;Im, Gyeong-Il;Jeong, Pyeong-Rim
    • Parasites, Hosts and Diseases
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    • v.29 no.3
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    • pp.293-310
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    • 1991
  • Clonorchis sinensis is a common parasite of man in Korea. Researches on the specific antigens of C. sinensis would be valuable not only because those elucidate the molecular characteristics of this fluke but also because it is applicable to immunodiagnosis. Although many monoclonal antibodies have been used in the field of parasite immunology, few articles on monoclonal antibodies against C. sinensis have been published so far. The aim of this study was to analyse C. sinensis antigens recognized by monoclonal antibodies, and to set up ELISA-inhibition test using C. sinensis specific monoclonal antibodies for improved specificity of immunodiagnostic tests. By fusion between spleen cells of the mice immunized with C. sinensis water-soluble crude adult worm antigens and plasmacytoma cells of mouse origin, 29 hybridoma clones secreting anti-C. sinensis monoclonal antibodies were made, and 8 clones among those were found specific. After cell cloning, isotypes of 6 selected specific monoclonal anti- bodies were determined to be IgGl, IgG2b and IgA. Four exposed antigenic determinants of natural infection were recognized by different specific monoclonal antibodies. By enzyme-immunoelectrotransfer blot, 10 KD, 34 KD antigenic determinants were found to be reacted with CsHyb 0714-20, CsHyb 0605-10 monoclonal antibodies, respectively, The antigenic determinant recognized by CsHyb 0714-20 monoclonal antibody was revealed to be located at the surface and parenchyme of a parasite by indirect immunoauorescent antibody technique, and those reacted with CsHyb 0605-10, CsHyb 0714-25 monoclonal antibodies were found at the parenchyme and intestine. The antigenic determinant reacted with CsHyb 0605-23 monoclonal antibody was found mainly around the uterine eggs. Four antigenic determinants recognized by specific monoclonal antibodies were all found to be present in the early eluted fractions of C. sinensis antigens separated by Sephadex G-200 gel filtration. By conventional ELISA, 75% of clonorchiasis cases were found positive, but 7.1% of normal controls and 37.5% of paragonimiasis cases showed false positives. However, by ELISA-inhibition test using C. sinensis specific monoclonal antibody (CsHyb 0605-23), 77.1% of clonorchiasis cases were found positive, and there were no false positives in normal controls or paragonimiasis cases, indicating 100% specificity. The ELISA- inhibition test using monoclonal antibodies was found to have same sensitivity and definitely high specificity in comparison with conventional ELISA for serodiagnosis of human clonorchiasis.

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Hair Heavy Metal Contents in Mentally Retarded Children III - In Association with Mercury - (정신지체아 두발 중 중금속 함량 III - 수은과의 관련성 -)

  • Han, Ki-Hwan;Jang, Bong-Ki;Park, Soon-Woo;Kim, Doo-Hie
    • Journal of Preventive Medicine and Public Health
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    • v.22 no.3 s.27
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    • pp.368-379
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    • 1989
  • The relationship between mercury level of hair and mental retardation was investigated. The 297 subjects with mental retardation were drawn from two schools providing special educational services, one, consisted of children living in an orphan home, another, children with parents. The 117 centre] subjects were drawn from whom had got average or above average academic achivement in a regular elementary school. Hair sample were taken from the nape of the neck and the mercury analysis was carried out on an atomic absorption spectrophotometer(IL 551). There was no relationship between mercury contents and age, and there was a statistically significant difference in mercury contents between male and female in the mentally retarded children living with parents. Children in the retarded group had significantly higher mercury contents compared with control group except the female group with parents. Also, the mercury levels in the retarded group living in an orphan home were significantly higher than that of the retarded group with parents. The concomitant diseases were Down's syndrome, epilepsy, cerebral palsy and autism. There were statistically significant differences in hair mercury levels in the cases of accompanying Down's syndrome and cerebral palsy in male and Down's syndrome and autism in female compared with the control group of the same sex. The most accompanying handicap was speech disturbance(40.7%) and the others were crippled, emotional disturbance etc. The percentages of double handicap were 66.7% among 6 persons exceeding 6ppm of their hair mercury contents. 10.4% among $3{\sim}6$ ppm and 15.7% among the group of 3ppm or less. The findings of this study suggest that the more opportunities of exposure to mercury in mentally retarded children may have occurred, so it can not be excluded the possibility of mercury as a contributing factor to mental retardation. Therefore, the causal relationship between mercury levels and mental retardation should be established through the examinations about their living environments, dietary pattern, eating habit etc.

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Extract from Eucheuma cottonii Induces Apoptotic Cell Death on Human Osteosarcoma Saos-2 Cells via Caspase Cascade Apoptosis Pathway (Eucheuma cottonii 추출물에 의한 인체 골육종암 Saos-2 세포의 자가사멸 유도)

  • Kang, Chang-Won;Kang, Min-Jae;Kim, Kyong Rok;Kim, Nan-Hee;Seo, Yong Bae;Kang, Keon-Hee;Kim, Sang-Ho;Kim, Gun-Do
    • Journal of Life Science
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    • v.26 no.2
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    • pp.147-154
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    • 2016
  • Osteosarcoma (OS) is the most common and malignant bone tumors. Although many types of resection surgery and experimental agents were developed, median survival and clinical prognosis are poorly investigated. Recently, several researches have reported that Eucheuma cottonii has potent as protective effects of coal dust-induced lung damage via inhibition of malondialdehyde (MDA) and oxidative stress in bronchoalveolar lavage fluids (BALF). However, anti-cancer effects and specific molecular mechanism of extract from Eucheuma cottonii (EE) has not been clearly studied yet. This study evaluated that anti-cancer potential of EE in human osteosarcoma Saos-2 cells. EE indicated cytotoxicity on Saos-2 cells in a dose-dependent manner. Morphological degradation and nucleic condensation were also observed under the EE treatment. However, it did not significantly affect on non-cancerous kidney HEK-293 cells under the same concentration which is shown cytotoxicity on Saos-2 cells. The phosphorylation of Fas-Associated Death Domain (FADD) and expression of cleaved caspase-8, -7 and -3 were upregulated in a dose-dependent manner. In immunofluorescence staining, expression level of Fas and cleaved PARP were upregulated by EE treatment. Furthermore, treatment of EE induces upregulation of sub G1 phase by flow cytometry analysis. The results demonstrated that EE has a therapeutic potential against osteosarcoma via FADD mediated caspase cascade apoptosis signal pathway.

Decrease of Aflatoxin M1 Level in Raw Cow’s Milk using the Hazard Analysis and Critical Control Points (HACCP) System (HACCP 제도에 의한 우유의 아플라톡신 M1의 저감화)

  • Kim, Ki-Hwan;Nam, Myoung Soo
    • Journal of Life Science
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    • v.26 no.2
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    • pp.190-197
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    • 2016
  • Aflatoxin M1 can be produced in cow’s milk when cows eat contaminated produce. Milk is a major source of food for infants and for children who have a weak level of immunity, and the detection of Aflatoxin M1 for risk assessment is necessary in order to reduce the amount of it in milk. In this study, the Aflatoxin M1 level was monitored for one year in raw milk samples obtained from Chungnam Province, Korea. The milk samples were divided into three categories: 1. milk samples from a standard general farm, 2. milk samples from a HACCP controlled farm, and 3. milk samples from the supply of Aflatoxin M1 reduced fodder. The average concentrations of Aflatoxin M1 in milk were 0.023±0.005 ug/l for the standard general farm, 0.017±0.004 ug/l for the HACCP controlled farm, and 0.013±0.003 ug/l for the supply of Aflatoxin M1 reduction fodder. Milk collected from the supply of Aflatoxin M1 reduction fodder had the lowest level of Aflatoxin M1. However, when efficiency and economic aspects are considered the most effective way of reducting Aflatoxin M1, could be taking milk from the HACCP controlled farm and implementing good feed management. Institutional support from the government, careful management of dairy farming, and a strict farm sanitation program are required in order to lower the level of Aflatoxin M1 in milk.

Determination of plasma C16-C24 globotriaosylceramide (Gb3) isoforms by tandem mass spectrometry for diagnosis of Fabry disease (패브리병(Fabry) 진단을 위한 혈장 중 Globotriaosylceramide (Gb3)의 탠덤매스 분석법 개발과 임상 응용)

  • Yoon, Hye-Ran;Cho, Kyung-Hee;Yoo, Han-Wook;Choi, Jin-Ho;Lee, Dong-Hwan;Zhang, Kate;Keutzer, Joan
    • Journal of Genetic Medicine
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    • v.4 no.1
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    • pp.45-52
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    • 2007
  • Purpose : A simple, rapid, and highly sensitive analytical method for Gb3 in plasma was developed without labor-ex tensive pre-treatment by electrospray ionization MS/ MS (ESI-MS/MS). Measurement of globotriaosy lceramide (Gb3, ceramide trihex oside) in plasma has clinical importance for monitoring after enzyme replacement therapy in Fabry disease patients. The disease is an X-linked lipid storage disorder that results from a deficiency of the enzyme ${\alpha}$-galactosidase A (${\alpha}$-Gal A). The lack of ${\alpha}$-Gal A causes an intracellular accumulation of glycosphingolipids, mainly Gb3. Methods : Only simple 50-fold dilution of plasma is necessary for the extraction and isolation of Gb3 in plasma. Gb3 in diluted plasma was dissolved in dioxane containing C17:0 Gb3 as an internal standard. After centrifugation it was directly injected and analyzed through guard column by in combination with multiple reaction monitoring mode of ESI-MS/MS. Results : Eight isoforms of Gb3 were completely resolved from plasma matrix. C16:0 Gb3 occupied 50% of total Gb3 as a major component in plasma. Linear relationship for Gb3 isoforms w as found in the range of 0.001-1.0 ${\mu}g$/mL. The limit of detection (S/N=3) was 0.001 ${\mu}g$/mL and limit of quantification was 0.01 ${\mu}g$/mL for C16:0 Gb3 with acceptable precision and accuracy. Correlation coefficient of calibration curves for 8 Gb3 isoforms ranged from 0.9678 to 0.9982. Conclusion : This quantitative method developed could be useful for rapid and sensitive 1st line Fabry disease screening, monitoring and/or diagnostic tool for Fabry disease.

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A Review of Service Innovation Research: A Comparison of Domestic and International Research Papers (서비스혁신 연구 동향: 국내 및 해외 주요 학술지를 중심으로)

  • Ryu, Hyun-Sun
    • Asia pacific journal of information systems
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    • v.24 no.4
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    • pp.577-610
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    • 2014
  • Although service innovation is not a new concept, innovation research in general tends to focus on technological innovation by manufacturing firms. With this view, innovation studies focus on product(e.g., goods) and process(e.g., product systems) innovation, ignoring service innovation and its inherent opportunities. Since major economy has been transformed to service economy, service innovation is considered a new effective way to sustain and gain a competitive advantage. Service innovation is no longer regarded merely as a side activity to product innovation; it has become a main research topic in its own right, accompanied by an increasing focus on services. While the number of service innovation studies has increased dramatically in the past 30 years in international research, few studies have been performed in domestic studies because domestic service innovation research began from the middle of 2000. In addition, there are no comprehensive literature reviews describing the evolution of service innovation research in both international and domestic studies because of the heterogeneities of service industry and multidiscipline characteristics of service innovation studies. To bridge this research gap, the purpose of this paper is to perform an extensive literature review and synthesis to enable a critical review of extant research on service innovation and trace its evolution, which will establish a foundation for further studies. By reviewing 169 articles (136 international papers; 33 domestic papers) published between 2000 and 2014 (in past 15 years), primarily in leading service, innovation and management information systems journals, this study analyzes the progression of service innovation research according to the four aspects such as number of studies, topics, methodologies and target industries. Overall, the view of service innovation has evolved, from a complement of traditional product innovation to a multidimensional, all-encompassing concept that entails several functions, both within and outside the firms. The results showed that domestic research still stays at the formation phase of service innovation studies although international research is in the maturity or multidimensional phase. We found increasing recent activities pertaining to service innovation, resulting from the increasing interest in services innovation across various industries and the links of new topics to the service innovation concept in both international and domestic studies. However, the main focus of service innovation research showed a different propensity between international and domestic studies: the former mainly focuses on a much more diversified pattern, emphasizing the linkages between service innovation and business strategy while the latter mainly focuses on the service innovation process(system) and service design. In addition, there are many case studies in domestic studies while many empirical studies in international studies. Domestic studies should increases the understanding of the interplay between service innovation and product innovation within manufacturing firms. Furthermore, rather than focusing on intrinsic distinctions between service innovation and product innovation, researchers should strive to develop and conceptualize service innovation in domestics studies. The present research also provides useful implications for practitioners. First, this study contributes to expand the current understanding of service innovation research by performing an extensive literature review. Second, tracing and comparing the progression and trends of service innovation research between international and domestic studies, this study showed the similarities and differences between them, which provide practical guidance on future research directions and research agenda. Third, this study performed literature review establishing the analysis system in the initial stage and using them to analyze articles, which is leading to explain the research review of service innovation more systematically and objectively. Finally, this study suggests the domestic researchers their future interests and topics of service innovation research.

Enhancement of Immunostimulation by Fractionation of Active Polysaccharide from Fermented Ginseng with Phellinus linteus Mycelium in Solid Culture (면역활성 증진을 위한 수삼의 상황버섯 균사체 고체배양으로 조제된 수삼발효물로부터 활성 다당류 분획)

  • Kim, Hoon;Yoon, Hyun-Seok;Jeong, Jae-Hyun;Jeong, Heon-Sang;Hwang, Jong-Hyun;Yu, Kwang-Won
    • Korean Journal of Food Science and Technology
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    • v.42 no.2
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    • pp.223-232
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    • 2010
  • Crude polysaccharide (WG-PL-CP) was fractionated from fermented ginseng with Phellinus linteus in solid culture to enhance the immunostimulation of ginseng. WG-PL-CP produce three active polysaccharide-rich fractions (WG-PL-CP-II, III, and IV) on DEAE-Sepharose CL-6B ($Cl^-$ form). WG-PL-CP-III displayed higher mitogenic activity (1.98-fold of the saline control at $100\;{\mu}g/mL$) than did WG-CP-III or PL-CP-III (1.60- or 1.65-fold, respectively), and potent intestinal immune system modulating activity through Peyer's patch was obtained by WG-PL-CP-IV only (1.56-fold). Meanwhile, WG-PL-CP-II and III significantly enhanced macrophage stimulating activity (2.01- and 1.94-fold) compared to WG-CP-II and III (1.73- and 1.66-fold) or PL-CP-II and III (1.79- and 1.72-fold). In addition, WG-PL-CP-III and IV mainly contained neutral sugar (73.5 and 67.3%) and uronic acid (23.2 and 24.6%). Component sugar analysis also showed that WG-PL-CP-III consisted mainly of uronic acid as well as the neutral sugars Glc, Ara, Gal, Rha and Xyl (molar ratio of 0.81:1.00:0.49:0.42:0.28:0.20), whereas WG-PL-CP-IV was mainly comprised of uronic acid, Ara, Rha, Gal, Xyl and Glc (1.00:0.75:0.69:0.63:0.42:0.34). Therefore, it is assumed that these active polysaccharides play an important role in enhancing the immunostimulation of fermented ginseng with P. linteus in solid culture.

Development of Detection Method for Niphon spinosus, Epinephelus bruneus, and Epinephelus septemfasciatus using 16S rRNA Gene (16S rRNA를 이용한 다금바리, 자바리, 능성어 판별법 개발)

  • Park, Yong-Chjun;Jung, Yong-Hyun;Kim, Mi-Ra;Shin, Joon-Ho;Kim, Kyu-Heon;Lee, Jae-Hwang;Cho, Tae-Yong;Lee, Hwa-Jung;Lee, Sang-Jae;Han, Sang-Bae
    • Korean Journal of Food Science and Technology
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    • v.45 no.1
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    • pp.1-7
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    • 2013
  • Niphon spinosus, Epinephelus bruneus, and Epinephelus septemfasciatus are involved in the Perciformes Order and Serranidae Family. When E. bruneus and E. septemfasciatus are fully grown, the striped pattern on the body gradually disappears. Therefore, morphological classification of adult fishes is quite difficult to identify the differences to N. spinosus. In this study, we investigate the method to differentiate those using PCR. To design the primers, 16S rRNA region of N. spinosus, E. bruneus, and E. septemfasciatus registered in the GeneBank (www.ncbi.nlm.nih.gov) have been used and for the analysis, Bio Edit ver. 7.0.9.0 was used. As a result, it was design NS-003-F/NS-005-R (136 bp), EB-001-F/EB-002-R (181 bp), and ES-001-F/ES-001-R (123 bp) primers for the differentiation of each 3 different fishes. Therefore, the species-specific primer sets would be a useful tool for scientific and speedy differentiation against the illegal distribution for consumer protection.

Change of Fragrant Components by Flowering Stages in Lilium Oriental Hybrid 'Casa Blanca' (오리엔탈 나리 '카사블랑카'의 개화단계별 향기성분 변화)

  • Rho, A Ran;Pak, Chun Ho
    • Horticultural Science & Technology
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    • v.19 no.2
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    • pp.163-169
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    • 2001
  • This experiment was conducted to find out suitable extraction time of available fragrant component in Lilium Oriental Hybrida 'Casa Blanca' based on qualitative and quantitative variation of its fragrant component in its flowering stages. The content of essential oil in its flowering stages increased with the progress of flowering stages except flower bud stage and it had the most oil content in its stage after full bloom. The analysis of the essential oil in Lilium Oriental Hybrida 'Casa Blanca' in its flowering stages shows that its main constituents were farnesol (18.96%), benzyl salicylate (13.81%), butyl-hydroxy toluene (12.87%), geranyl linalool isomer (7.46%), isoeugenol (7.17%) in its each stage. Benzenoids had much content at half bloom stage and full bloom stage while fatty acid derivatives had much content at initial flowering stage and after flowering stage. Most benzenoids such as butyl-hydroxy toluene and isoeugenol, which are some of main constituents of Lilium Oriental Hybrida 'Casa Blanca' had strong antioxidant effect. So they were expected to be used as antioxidant agents for food, feed, vegetable oil. The content of monoterpene compounds like geranyl linalool isomer increased at its later stage. Sesquiterpene such as farnesol, which is main component of lily existed only at full bloom stage. Therefore farnesol and geranyl linalool isomer of Lilium Oriental Hybrida 'Casa Blanca' are expected to be used for important spices with good fragrance. The suitable extraction time for the usable main constituents of Lilium Oriental Hybrida 'Casa Blanca' is as follows; farnesol and benzyl salicylate: at full bloom stage, butyl-hydroxy toluene: at half bloom stage, geranyl linalool isomer and isoeugenol: at stage after full bloom. Finally there was variation in essential oil components and contents of Lilium Oriental Hybrida 'Casa Blanca' in its flowering stages and they are expected to be used usefully for flavor industry, food industry, aromatherapy, when they are extracted at their suitable extraction time.

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