• Title/Summary/Keyword: HPLC Chromatography

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Preparation and Release Behavior of Atorvastatin Calcuim - Encapsulated Polyoxalate Microspheres (아토르바스타틴 칼슘을 함유한 폴리옥살레이트 미립구의 제조 및 방출거동)

  • Lee, Cheon Jung;Kim, Su Young;Lee, Hyun Gu;Yang, Jaewon;Park, Jin Young;Cha, Se Rom;Lim, Dong-Kwon;Lee, Dongwon;Khang, Gilson
    • Polymer(Korea)
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    • v.38 no.5
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    • pp.656-663
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    • 2014
  • Atorvastatin calcium-loaded polyoxalate (POX) microspheres were prepared by an emulsion solvent-evaporation/ extraction method of oil-in-oil-in-water ($O_1/O_2/W$) for sustained release. We investigated the release behavior according to initial drug ratio, molecular weight ($M_w$) and concentration of POX and concentration of emulsifier. The microsphere was characterized on the surface, the cross-section morphology and the behavior of atorvastatin calcium release for 10 days by scanning electron microscopy (SEM) and high performance liquid chromatography (HPLC). The analysis of crystallization was analyzed to use X-ray diffraction (XRD), differential scanning calorimeter (DSC) and Fourier transform infrared (FTIR). These results showed that the release behaviors can be controlled by preparation conditions.

Identification of a Neolignan Glycoside from the Pine Tree, Pinus densiflora Showed Antithrombotic Activity (Pinus densiflora 유래의 항트롬빈 활성을 나타내는 Neolignan Glycoside의 동정)

  • Seo, Min-Jeong;Kang, Byoung-Won;Jeong, Yong-Kee
    • Journal of Life Science
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    • v.24 no.8
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    • pp.873-879
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    • 2014
  • The constituents from the needles of the pine tree, Pinus densiflora, were purified and investigated for antithrombotic activity. The needles were initially extracted three times with 70% ethanol, and the extract was sequentially fractionated with chloroform and n-butanol. The aqueous layer formed after n-butanol fractionation was subjected to purification by medium pressure and high pressure liquid chromatography. The two neolignans, 2, 3-dihydro-7-hydroxyl-3-hydroxymethyl-2-(4'-hydroxyl -3-methoxyphenyl)-5-benzofuranpropanol-3-O-${\alpha}$-rhamnopyranoside (a neolignan glycoside) and 2, 3-dihyro-3-hydroxymethyl-7-methoxy-2-(4'-hydroxyphenyl-3'-methoxy)-5-benxofuran propanol 4'-O-${\alpha}$-rhamnopyranoside (icariside $E_4$) were identified by $^1H$ and $^{13}C$ NMR spectra. The effect of the purified compounds, the neolignan glycoside and icariside $E_4$ on thrombin inhibition were investigated by measuring thrombin clotting time in plasma. As a result, the clotting of the neolignan glycoside was delayed four times compared to that of icariside $E_4$. In addition, an analysis of the inhibition effect by changing the concentration showed that the clotting time was delayed in accordance with an increase in the concentration of the neolignan glycoside. Furthermore, we examined the interaction of thrombin and fibrinogen to clarify the action mechanism. As a result, the delay of clotting time in the response of thrombin and pure fibrinogen may indicate that neolignan glycosides inhibit the thrombin action in a direct manner, leading to the suppression of fibrin generation.

Metabolic engineering of Vit C: Biofortification of potato

  • Upadhyaya, Chandrama P.;Park, Se-Won
    • Proceedings of the Plant Resources Society of Korea Conference
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    • 2010.10a
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    • pp.14-14
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    • 2010
  • Vitamin C (ascorbic acid) is an essential component for collagen biosynthesis and also for the proper functioning of the cardiovascular system in humans. Unlike most of the animals, humans lack the ability to synthesize ascorbic acid on their own due to a mutation in the gene encoding the last enzyme of ascorbate biosynthesis. As a result, vitamin C must be obtained from dietary sources like plants. In this study, we have developed two different kinds of transgenic potato plants (Solanumtuberosum L. cv. Taedong Valley) overexpressing strawberry GalUR and mouse GLoase gene under the control of CaMV 35S promoter with increased ascorbic acid levels. Integration of the these genes in the plant genome was confirmed by PCR and Southern blotting. Ascorbic acid(AsA) levels in transgenic tubers were determined by high-performance liquid chromatography(HPLC). The over-expression of these genes resulted in 2-4 folds increase in AsA intransgenic potato and the levels of AsA were positively correlated with increased geneactivity. The transgenic lines with enhanced vitamin C content showed enhanced tolerance to abiotic stresses induced by methyl viologen(MV), NaCl or mannitol as compared to untransformed control plants. The leaf disc senescence assay showed better tolerance in transgenic lines by retaining higher chlorophyll as compared to the untransformed control plants. Present study demonstrated that the over-expression of these gene enhanced the level of AsA in potato tubers and these transgenics performed better under different abiotic stresses as compared to untransformed control. We have also investigated the mechanism of the abiotic stress tolerance upon enhancing the level of the ascorbate in transgenic potato. The transgenic potato plants overexpressing GalUR gene with enhanced accumulation of ascorbate were investigated to analyze the antioxidants activity of enzymes involved in the ascorbate-glutathione cycle and their tolerance mechanism against different abiotic stresses under invitro conditions. Transformed potato tubers subjected to various abiotic stresses induced by methyl viologen, sodium chloride and zinc chloride showed significant increase in the activities of superoxide dismutase(SOD, EC 1.15.1.1), catalase, enzymes of ascorbate-glutathione cycle enzymes such as ascorbate peroxidase(APX, EC 1.11.1.11), dehydroascorbate reductase(DHAR, EC 1.8.5.1), and glutathione reductase(GR, EC 1.8.1.7) as well as the levels of ascorbate, GSH and proline when compared to the untransformed tubers. The increased enzyme activities correlated with their mRNA transcript accumulation in the stressed transgenic tubers. Pronounced differences in redox status were also observed in stressed transgenic potato tubers that showed more tolerance to abiotic stresses when compared to untransformed tubers. From the present study, it is evident that improved to lerance against abiotic stresses in transgenic tubers is due to the increased activity of enzymes involved in the antioxidant system together with enhanced ascorbate accumulated in transformed tubers when compared to untransformed tubers. At moment we also investigating the role of enhanced reduced glutathione level for the maintenance of the methylglyoxal level as it is evident that methylglyoxal is a potent cytotoxic compound produced under the abiotic stress and the maintenance of the methylglyoxal level is important to survive the plant under stress conditions.

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Real-Time Monitoring of Mitochondrial ATP Synthesis and Hydrolysis by Surface Infrared Spectroscopy

  • Yamaguchi, Ryo-Taro;Hirano-Iwata, Ayumi;Aonuma, Yuki;Yoshimura, Yuya;Shinohara, Yasuo;Kimura, Yasuo;Niwano, Michio
    • Proceedings of the Korean Vacuum Society Conference
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    • 2013.02a
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    • pp.108-109
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    • 2013
  • Mitochondria play key roles in the production of cell's energy. Their dominant function is the synthesis of adenosine 5'-triphosphate (ATP) from adenosine diphosphate (ADP) and phosphate (Pi) through the oxidative phosphorylation. Evaluation of drug-induced mitochondrial toxicity has become increasingly important since mitochondrial dysfunction has recently been implicated in numerous diseases including cancer and diabetes mellitus. Mitochondrial functions have been monitored via oxygen consumption, mitochondrial membrane potential, and more importantly via ATP synthesis since ATP synthesis is the most essential function of mitochondria. Various analytical methods have been employed to investigate ATP synthesis in mitochondria, including high performance liquid chromatography (HPLC), bioluminescence technique, and pH measurement. However, most of these methods are based on destructive analysis or indirect monitoring through the enzymatic reaction. Infrared absorption spectroscopy (IRAS) is one of the useful techniques for real-time, label-free, and direct monitoring of biological reactions [1,2]. However, the strong water absorption requires very short path length in the order of several micrometers. Transmission measurements with thin path length are not suitable for mitochondrial assays because solution handlings necessary for evaluating mitochondrial toxicity, such as rapid mixing of drugs and oxygen supply, are difficult in such a narrow space. On the other hand, IRAS in the multiple internal reflection (MIR) geometry provides an ideal optical configuration to combine solution handling and aqueous-phase measurement. We have recently reportedon a real-time monitoring of drug-induced necrotic and apoptotic cell death using MIR-IRAS [3,4]. Clear discrimination between viable and damaged cells has been demonstrated, showing a promise as a label-free and real-time detection for cell-based assays. In the present study, we have applied our MIR-IRAS system to mitochondria-based assays by monitoring ATP synthesis in isolated mitochondria from rat livers. Mitochondrial ATP synthesis and hydrolysis were in situ monitored with MIR-IRAS, while dissolved oxygen level and solution pH were simultaneously monitored with O2 and pH electrodes, respectively. It is demonstrated that ATP synthesis and hydrolysis can be monitored by the IR spectral changes in phosphate groups in adenine nucleotides and MIR-IRAS is useful for evaluating time-dependent drug effects of mitochondrial toxicants.

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The Effects of Puerariae Radix Catechins Administration on Liver Function in Carbon Tetrachloride-Treated Rats (사염화탄소를 투여한 흰쥐에 있어서의 간 기능에 미치는 칡 카테킨의 효과)

  • 이치호;한석현;김종배;민상기
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.24 no.5
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    • pp.713-719
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    • 1995
  • These studies were carried out to investigate the effects of Puerariae radix catechins(PRC) administration on the biochemical parameters of liver function in liver of carbon tetrachloride(CCl4)-treated rats. Thirty six healthy Sprague-Dawley rats weighing about 120g were used for this experiment and divided intot he following 3 groups : normal control group(NCON), $CCl_4$ control group(CCON), PRC treated group(PRC). Fifty percent $CCl_4$ in oil was administered(I.P.) by 2ml per kg body weight two times a week for 3 weeks. PRC treated groups were administered orally at the leaves of 1% per day in distilled water for 8 weeks. Lipid hydroperoxides were analyzed by using chemiluminescence-high performance liquid chromatography(CL-HPLC) method as a phosphatidylcholine hydroperoxide value(PCOOH) in liver tissues. $CCl_4$ treatment significantly(p<0.05) resulted in an increase in GPT & GOT activities and liver hydroperoxide values comparing with those of the untreated control, while administration of PRC to the $CCl_4-treated$ rats significantly(p<0.001) decreased GPT & GOT activities and liver hydroperoxide value. Their ultrastructual changes of hepatocellular organelles were shown to clarify the morphologic nature of protective effects of PRC on hepatocytic injuries. $CCl_4$ treatment observed to change the ultrastructual nature of outer membrane of hepatocytes. However, the hepatic changes on PRC treatment to $CCl_4$ group was not found. PRC administration may inhibit the formatiion of liver lipid hydroperoxides in vivo and were very effective in recovering the liver function in $CCl_4-treated$ rats.

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Bioequivalence of RoxithrinTM Tablet to RulidTM Tablet (Roxithromycin 150 mg) (루리드 정(록시스로마이신 150 mg)에 대한 록시스린 정의 생물학적동등성)

  • Joung, Sun-Koung;Lee, Yun-Young;Cho, Tae-Seob;Kim, Ho-Hyun;Lee, Ye-Rie;Lee, Kyung-Ryul;Lee, Hee-Joo
    • Journal of Pharmaceutical Investigation
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    • v.34 no.3
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    • pp.209-214
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    • 2004
  • A bioequivalence study of $Roxithrin^{TM}$ tablet (Kukje Pharma. Ind. Co., Ltd.) to $Rulid^{TM}$ tablet (Han Dok Pharma. Ind. Co., Ltd.) was conducted according to the guidelines of Korea Food and Drug Administration (KFDA). Twenty four healthy male Korean volunteers received each medicine at the roxithromycin dose of 300 mg in a $2{\times}2$ crossover study. There was a one-week wash-out period between the doses. Plasma concentrations of roxithromycin were monitored by a high-performance liquid chromatography for over a period of 36 hours after drug administration. $AUC_t$ (the area under the plasma concentration-time curve from time zero to 36 hr) was calculated by the linear trapezoidal rule method. $C_{max}$ (maximum plasma drug concentration) and $T_{max}$ (time to reach $C_{max}$) were compiled from the plasma concentration-time data. Analysis of variance was carried out using logarithmically transformed $AUC_t$ and $C_{max}$. No significant sequence effect was found for all of the bioavailability parameters indicating that the cross-over design was properly performed. The 90% confidence intervals of the $AUC_t$ ratio and the $C_{max}$ ratio for $Roxithrin^{TM}/Rulid^{TM}$ were 1.00 - 1.13 and 0.98 - 1.10, respectively. These values were within the acceptable bioequivalence intervals of 0.80 - 1.25. Thus, our study demonstrated the bioequivalence of $Roxithrin^{TM}$ and $Rulid^{TM}$ with respect to the rate and extent of absorption.

Establishment of analytical method of methiozolin and dissipation in golf course's green (Methiozolin의 분석법 개발 및 골프장 그린에서의 잔류소실특성)

  • Jo, Hyeong-Wook;Hwang, Kyu-won;Hwang, Ki-Hwan;Moon, Joon-Kwan
    • Journal of Applied Biological Chemistry
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    • v.59 no.4
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    • pp.331-336
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    • 2016
  • Analytical methods for methiozolin in soil, water and turfgrass were established and dissipation patterns of methiozolin in soil and turfgrass were elucidated. Analysis was done using a high performance liquid chromatography with an ultra violet detector at the wavelength of 280 nm after extraction with acetone, liquid-liquid partition with dichloromethane, and a solid phase extraction purification. Limit of determination and Limit of quantitation were 1.0, 0.5, 1.0 ng, and 0.001, 0.1, 0.01 mg/kg for water, turfgrass, and soil, respectively. Recovery rates of methiozolin from soil, water, and turfgrass were ranged 87.5~111.3, 92.8~97.4, and 78.2~98.5 %, respectively. The turfgrass and soil samples were collected at 0, 1, 4, 7, 14, 30, 45, and 60 after spray on green area in golf course. Residues of methiozlolin were not translocated to lower soil layer but detected only in turfgrasses and root area of turfgrass. Half-lives of methiozolin in turfgrass were 10.7 days and 8.8 days in soil from root area.

Dissipation characteristics of mandipropamid and thiamethoxam for establishment of pre-harvest residue limits in lettuce (상추의 생산단계 잔류허용기준 설정을 위한 농약 Mandipropamid 및 Thiamethoxam의 잔류소실특성 연구)

  • Yang, Seung-Hyun;Lee, Jae-In;Choi, Hoon
    • Journal of Applied Biological Chemistry
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    • v.63 no.3
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    • pp.267-274
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    • 2020
  • The dissipation characteristics and kinetics of fungicide mandipropamid and insecticide thiamethoxam in lettuce under greenhouse conditions were investigated at three different lettuce-growing fields for estimating the pre-harvest residue limits (PHRLs). The analytical methods were fully validated for the quantitation of pesticide residues using High-Performance Liquid Chromatography-Photo Diode Array detector or Ultraviolet-Visible Detector and applied to real samples. The lettuces suitable for shipment were harvested during 10 days including pre-harvest interval after treatment at the recommended dose by safe-use guidelines. The initial mean residues in different fields were 6.68-17.87 and 4.96-8.31 mg/kg for mandipropamid and thiamethoxam, respectively, which decreased to 16-54 and 14-44% in 10 days. The clothianidin, a metabolite of thiamethoxam, was detected in <0.02 to 0.37 mg/kg. The dissipation of both pesticides followed first-order kinetics over a period of 10 days after application. Based on the residue data, the mean dissipation rate constant (λ) and biological half-lives (T1/2) were estimated to be -0.1060 and 6.5 days of mandipropamid and -0.1236 and 5.6 days of thiamethoxam. The PHRLs for lettuce on the 10th and 5th day before harvesting were calculated to be 63.24 and 43.56 mg/kg for mandipropamid, and 44.66 and 25.88 mg/kg for thiamethoxam, with -0.0746 and -0.1091 of the upper 95% confidence intervals of dissipation rate constant, respectively. This work would be useful as guidance for adjusting the shipment date and contribute to stabilizing the income of farmers in Korea.

온라인 협동조합의 공생마케팅 전략-웹기반 사진앨범협동조합 (주)와이드스쿨 사례-

  • 김창호
    • Distribution Business Review
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    • no.3
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    • pp.155-170
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    • 2003
  • 본 연구는 기본적으로 온라인과 오프라인의 통합마케팅을 절명하고 이에 관한 경험적 사례를 개발하기 위한 목적으로 진행되었다. 앨범서비스 영역의 공생적 기반 위에 전개되는 온 -오프라인의 경험적 사례를 개발하고 바람직한 마케팅방향 방향을 제시하였다. 본 연구는 문헌연구와 사례연구를 병행하여 연구를 진행하였다. 사례는 인터넷 기반의 앨범서비스를 제공하기 위한 (주)와이드스쿨이다. 온-오프라인의 협력적 통합마케팅의 전략을 전개하기 위해서는 무엇보다도 온 -오프라인의 뚜렷한 목표를 절정하고 성장방향에 대한 비전을 공유하고 나아가 온 -오프라인의 사명을 감당하는 것이다. 즉, 실천적으로는 \circled1 항상 고객 (customer)기반의 의사결정을 이루며 \circled2 철저한 협력적 돕는 경쟁(competition) 의식과 \circled3 구성원 자신의 일에 대한 자신감(confidence)을 지니고 \circled4 실천을 위한 용기(courage)를 가지고 \circled4 혁신하여 변화(change)를 선도하는 것이다. 온라인(on line)으로 표현되는 인터넷환경은 모든 영역에 변화를 요구하고 있다. 온라인에 관한 연구는 크게 온라인시장의 경쟁(competition)에 관한 연구와, 온라인 소비자(consumer)에 관한 연구 그리고 온라인 시장 참가기업(company)에 관한 연구로 구분된다(이석규 ; 2001). 이중 기업에 관한 연구의 중심에는 e-biz의 수익모텔에 관한 연구가 주류를 이루고 았다(David et al, 1999) 특히 오프라인기업의 경우 어떠한 형태방법으로 온라인 환경에 부응하며 기존의 마케팅활동과 연계할 것인가는 매우 중요한 문제다. 즉 기존의 오프라인기업이 온라인도구변화에 적응하고 이를 전략적으로 활용하기 위해서는 무엇보다도 오프라인과 온라인의 통합에 관한 형태와 전략 등을 명확히 이해하고 적용하는 것이 중요하다. 개수가 감소하는 것과는 상당히 다른 분포이다. 따라서 우리의 관측 결과는 2001년 사자자리 유성우의 극대 시간 전후 2시간에 적어도 0등급 이하의 밝은 유성이 상대적으로 많이 발생하였을 것으로 해석된다. 이런 밝은 유성의 빈도는 유성우 특성 연구에 중요한 의미를 가진다. 그러나 표준성만을 이용해 결정된 유성 등급은 유성의 지속 시간에 대한 불확실성과 전천 카메라 감응도의 비선형성에 의한 불확실성을 내포하고 있음을 지적해 둔다.umn chromatography)를 사용하였고 일련의 정제 과정을 통하여 배양액 중의 L-lactic acid 정제 수율은 약 85% 정도로 나타났으며 HPLC로 분석한 결과 99.7%의 순도를 확인할 수 있었다.경향을 나타내며 유입휫수와 $Dst_{min}$ 사이에는 높은 상관관계(0.83)가 있었다. 둘째, 주상기간 중 자기폭풍의 크기가 클수록 플럭스 비 ($f_{max}/f_{ave}$는 대체로 증가하는 경향을 나타냈다. 그리고 75~113keV 에너지 채널에서의 $Dst_{min}$ 값과 플럭스 비의 상관계수는 0.74로서 가장 높았으며 나머지 에너지 채널 역시 비교적 높은 상관관계를 나타냈다. 셋째, 주상기간 중 총 에너지 유입률 지수와 $Dst_{min}$ 사이에 높은 상관관계가 확인되었다. 특히 환전류를 구성하는 주요 입자의 에너지 영역(75~l13keV)에서 가장 높은(0.80) 상관계수를 기록했다. 넷째, 회복기 중에 일어나는 입자들의 유입은 자기폭풍의 지속시간을 연장시키는 경향을 보이며 큰 자기폭풍일수록 현저했다. 주상에서 관측된 이러한 특성은 서브스톰 확장기 활동이 자기폭풍의 발달과 밀접한 관계가 있음을 시사한다.se that were all low in two aspects, named "the Nonsignificant group".

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Antimicrobial Activity of Some Plants Containing Allelochemicals (Allelochemicals 함유 식물의 항균 효과)

  • 이호준;김용옥
    • The Korean Journal of Ecology
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    • v.22 no.1
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    • pp.51-58
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    • 1999
  • Korean native plants and naturalized plants were analyzed for allelochemicals, and their antimicrobial effects were studied. The difference in soluble solid contents between Korean native plants and naturalized species was not significant, and the Korean native plant, Solanum nigrum showed the highest soluble solid content of 90 mg/ml. The ethanol extract of the Korean native plant, Solanum nigrum showed antifungal activity to Aspergillus phoenicis KCTC 1228, with a clear zone of 18 mm, and spore formation was not observed from the treatment. The naturalized plants Ambrosia artemisiifolia var. elatior and Erigeron canadensis showed inhibition of spore formation and the clear zones were at 24 mm and 22 mm, respectively. The clear zones of Aspergillus phoenicis KCTC 1228 treated with ethanol extrats of Phytolacca americana and Rudbeckia bicolor were 22 mm and 19 mm, respectively, and spore formation was observed from the treatment. The Korean native plant, Solanum nigrum and naturalized plants, Phytolacca americana and Ambrosia artemisiifolia var. elatior showed antimicrobial activity against Bacillus sphiaericus 2362, and Bacillus sphiaericus 2297, Bacillus thuringiensis var. subtilis and Baicillus thuringiensis var. cereus. The antimicrobial activity of Ambrosia artemisiifolia var. elatior showed the largest clear zone of 32 mm against Bacillus thuringiensis var. subtilis. In general, the more soluble the solid contents of the extracts, the greater were the antifungal and antimicrobial activities. The phenolic compounds from the Korean native plant, Solanum nigrum and the naturalized species, Phytolacca americana and Ambrosia artemisiifolia var. elatior were analyzed by high performance liquid chromatography. Three phenolic compounds including hydroquinone were identified in Solanum nigrum. In contrast, five and seven phenolic compounds were identified in Phytolacca americana and Ambrosia artemisiifolia var. elatior, respectively. The antifungal activity against Aspergillus phoenicis KCTC 1228 was found to be due to the coumaric and benzoic compounds.

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