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Quantitative Analysis of Atractylenolides I and III in Atractylodes japonica (백출의 Atractylenolide I과 Atractylenolide III의 함량분석)

  • Yun, Bo-Ra;Weon, Jin Bae;Lee, Bohyoung;Lee, Jiwoo;Eom, Min Rye;Ma, Choong Je
    • Korean Journal of Pharmacognosy
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    • v.44 no.1
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    • pp.53-59
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    • 2013
  • This study was conducted to determine the content of atractylenolides I and III in Atractylodes japonica. An established HPLC-DAD method was used to monitor contents of atractylenolides I and III in A. japonica obtained from Korea and China and compared with contents of A. macrocephala, A. chinensis, and A. lancea. Quantitative analysis of atractylenolides I and III was carried out on a Shiseido C18 column (S-5 ${\mu}m$, 4.6 mm I.D. ${\times}$ 250 mm) with gradient elution composed of acetonitile-water. The results show that the average content of atractylenolides I and III in A. japonica were 0.0954 and 0.1963%, respectively, and contents of A. lancea were higher than A. macrocephala, A. chinensis. In this study, we identified the differentiation of the quality of A. japonica from different species and collected locations and established content standard of atractylenolides I and III in A. japonica and this content standard was helpful to quality control of A. japonica.

Antifungal Activity of Eucalyptus-Derived Phenolics Against Postharvest Pathogens of Kiwifruits

  • Oh, Soon-Ok;Kim, Jung-A;Jeon, Hae-Sook;Park, Jong-Cheol;Koh, Young-Jin;Hur, Hyun;Hur, Jae-Seoun
    • The Plant Pathology Journal
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    • v.24 no.3
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    • pp.322-327
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    • 2008
  • Antifungal activities of natural substrances from Eucalyptus darlympleana, E. globules, E. gunnii and E. unigera were evaluated against postharvest pathogens of kiwifruits, Botrytis cinerea, Botryosphaeria dothidea, and Diaporthe actinidiae, to screen effective natural substances as an alternative to chemical fungicides. Methanol extract of the Eucalyptus trees showed strong antagonistic activity against the pathogenic fungi. Among them, E. unigera and E. darlympleana effectively inhibited mycelial growth of the pathogens. For chemical identification of the antifungal substances, the methanol extract of E. darlympleana leaves was successively partitioned with $CH_2Cl_2$, EtOAc, n-BuOH and $H_2O$. Among the fractions, $CH_2Cl_2$ and n-BuOH showed strong inhibitory activity of mycelial growth of the fungi. Five compounds were isolated from EtOAc and n-BuOH fractions subjected to $SiO_2$ column chromatography. Two phenolic compounds(gallic acid and 3,4-dihydroxybenzoic acid) and three flavonoid compounds(quercetin, quercetin-3-O-$\alpha$-L-rhamnoside, quercetin-3-O-$\beta$-glucoside) were identified by $^1H$-NMR and $^{13}C$-NMR spectroscopy. Among them, only gallic acid was found to be effective in mycelial growth and spore germination of B. cinerea at relatively high concentrations. The results suggest that gallic acid can be a safer and more acceptable alternative to current synthetic fungicides controlling soft rot decay of kiwifruit during postharvest storage.

Spalling Reduction Method of High Strength Reinforced Concrete Columns Using Fibers (섬유를 활용한 고강도 콘크리트기둥의 폭렬제어방안)

  • Yoo, Suk-Hyeong
    • Fire Science and Engineering
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    • v.23 no.4
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    • pp.7-12
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    • 2009
  • As the concrete strength increases the degree of damage caused by the spalling becomes more serious because of the permeability. It is reported that the polypropylene (PP) fiber has an important role in protecting concrete from spalling. However, the excessive usage of PP fiber would not useful in spalling control and would decrease the workability of ultra high strength concrete. The high-temperature behaviors of high-strength reinforced concrete columns with various dosage of PP fibers and three types of fire endurance fibers were observed this study. In results, the ratio of unstressed residual strength of columns, in case of concrete strength 60MPa, increases as the dosage of PP fiber increases from 0% to 0.2%, however, the effect of fiber dosage on residual strength of column barely changes above 0.2% and in case of concrete strength 120MPa, PVA fiber is the most suitable fire endurance fiber in accounting fire endurance performance and workability.

Antioxidative Activity of Browning Products Fractionated from Fermented Soybean Sauce (양조간장에서 분리한 갈색물질의 항산화성)

  • 최홍식;이정수;문갑순;박건영
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.22 no.5
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    • pp.565-569
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    • 1993
  • Antioxidative activity of browning product(BP) fractionated from fermented soybean sauce(SS) was studied during the oxidation process of linoleic acid mixture system. SSBP was a powder type product prepared from fermented soybean sauce by the fractionation through the Sephadex G-10 column and freeze drying of collected fraction. The aqueous model systems were used for the evaluation of antioxidative activity of SSBP during the oxidative reaction at $50^{\circ}C$ by the determination of peroxider and conjugated dienoic acid compounds. The linoleic acid mixture for the aqueous model systems was consisted of linoleic acid(64.6%), oleic acid(27.4%), and other acids in ethanolic phosphate buffer solution(pH 7.0). SSBP had a considerable antioxidative activity with the inhibition of formation of peroxides and conjugated dienoic acids during the autoxidation of linoleic acid mixtures in aqueous model systems. Antioxidative activity of SSBP was relatively higher than SS, however, lower than ${\alpha}-tocopherol$ and butylated hydroxyanisol. The antioxidative effect of SSBP was increased by the its concentrations from 0.05% to 0.5% in the oxidation reactions of aqueous model systems. Therefore, SSBP was considered as one of the potential natural antioxidants for the use of food products.

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Production of Mycelia and Water Soluble Polysaccharides from Submerged Culture of Lentinus lepideus in Bioreactor (생물반응기를 이용한 잣버섯(Lentinus lepideus)의 균사체 및 수용성 다당체 생산특성)

  • Ahn, Jin-Kwon;Ka, Kang-Hyeon;Lee, Wi-Young
    • The Korean Journal of Mycology
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    • v.35 no.1
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    • pp.37-42
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    • 2007
  • The mushroom Lentinus lepideus was used to produce mycelial as well as soluble polysaccharides in bioreactor cultures. To determine optimal submerged culture conditions, both growth characteristics and water soluble polysaccharides production were compared among four different types of bioreactor and culture conditions. For the production of mycelial biomass, the following bioreactors were proven to be effective in decreasing order: an external-loop type air-lift bioreactor (ETAB; 7g/l), a balloon type air bubble bioreactor (BTBB; 6.2g/l), a stirrer type bioreactor (STB; 6g/l), and a column type air bubble bioreactor (CTBB; 5g/l). Maxiaml production of water soluble exopolysaccharides (EPS; 0.62g/l) and endopolysaccharides (PPS; 7.7%) could also be obtained from BTBB. The mycelial biomass increased with increase in glucose concentration from 15g/l to 75g/l in the media. In contrast, PPS contents in the cells decreased with increase in glucose concentration in the media, showing the highest PPS content (7%) at 15g/l. Among different medium feeding types, fed-batch culture based on concentration control in media (10g/l) produced higher mycelia than fed-batch culture based on volume control of media (5.8g/l) or batch culture (3.4g/l). EPS production was also higher in fed-batch culture based on medium concentration control than that in other feeding types.

Fingerprint of Marker Substances in Gami-Honghwa-Tang(KH-19) by HPLC-DAD (High Performance Liquid Chromatography-Diode Array Detector(HPLC-DAD)에 의한 가미홍화탕 (KH-19)의 지문 분석)

  • Yu Young-Beob;Yoon Yoo-Sik;Cho Gi-Ho
    • The Journal of Korean Medicine
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    • v.25 no.3
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    • pp.45-54
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    • 2004
  • Objectives : This study was aimed to evaluate marker substances in Gami-Honghwa-Tang (KH-19) by high performance liquid chromatography-photodiode array detector (HPLC-DAD). Gami-Honghwa-Tang is composed of nine crude herbs, Rehmanniae Radix Preparata, Angelicae Gigantis Radix, Cnidii Rhizoma, Paeoniae Radix, Corni Fructus, Moutan Cortex Radicis, Lycii Fructus, Carthami Flos and Glycyrrhizae Radix. Methods : The separation was performed on an Aquasil C18 (4.6×250mm) column by gradient elution with 0.05% TFA in H2O - 0.05% TFA in acetonitrile (0 min 100:0, 20 min 90:10, 40 min 70:30, 60 min 50:50, 80 min 0:100, 90 min 100:0) as the mobile phase at a flow-rate of 1.0 ml/min with detection at 190-800nm. Also we examined the contents for bacteria, pesticide residue and harmful heavy metals. Results : HPLC-DAD was employed to determine the quantities and the qualities of several marker substances such as 5­hydroxymethyl-2-furaldehyde (5-HMF), paeonol, loganin, paeoniflorin, glycyrrhizin, and decursin in the KH-19. There were no bacterial contents, pesticide residues, or harmful heavy metals. Conclusions : We suggest these results could be a useful evidence for quality control of KH-19. This method permits fingerprints of selected individual marker substances from herbal prescriptions without derivatization, multiple purification steps, or lengthy separation times.

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Isolation of an Acinetobacter junii SY-01 Strain Producing an Extracellular Lipase Enantioselectively Hydrolyzing Itraconazole Precursor, and Some Properties of the Lipase

  • Yoon, Moon-Young;Shin, Pyong-Kyun;Han, Ye-Sun;Lee, So-Ha;Park, Jung-Keug;Cheong, Chan-Seong
    • Journal of Microbiology and Biotechnology
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    • v.14 no.1
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    • pp.97-104
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    • 2004
  • Water-sludge bacteria were screened to find a lipase enantioselectively hydrolyzing itraconazole precursor, which is well known as the starting material of antifungal drug agents. A bacterial strain was isolated and identified as Acinetobacter junii SY-01. After the strain was cultivated, the enzyme was purified 39.4-fold using ultrafiltration and gel filtration through a Sephadex G-100 chromatographic column and the activity yield was 34.9%. The molecular weight of the enzyme was about 40 kDa, as measured by SDS-PAGE, and the optimum pH was 7.0- 9.0 and stable at pH 6.0- 9.0. The optimum temperature was 45- $5^{\circ}C$, and 73% of the enzymes activity remained after incubation at 70% for 1 h. Enzyme activity was enhanced by gall powder, sodium deoxycholate, a cationic detergent Tween 80, and a non-ionic detergent Triton X-100, but was markedly inhibited by metal ions such as $Hg^{2+},Cu^{2+},Ni^{2+}/,Ca^{2+}$, and an anionic-surfactant sodium dodecylsulfate. The $K_{m}$ values for (R)- and (S)-enantiomers of the itraconazole precursor were 0.385 and 21.83 mM, respectively, and the $V_{max} values ($\mu$Mㆍmin^{-1}.)$ were 6.73 and 6.49, respectively. The acetyl group among the different acyl moieties of itraconazole precursor showed the highest enantioselectivity for the hydrolysis by the Acinetobacter junii SY-01 lipase, and the lipase from Acinetobacter junii SY-01 displayed better enantioselectivity than that of commercially available lipases and esterases.

Determination of isoquinoline alkaloids by UPLC-ESI-Q-TOF MS: Application to Chelidonium majus L.

  • Jeong, Won Tae;Lim, Heung Bin
    • Analytical Science and Technology
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    • v.30 no.6
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    • pp.379-389
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    • 2017
  • In this study, we set up an analytical method that can be used for rapid and accurate determination of representative isoquinoline alkaloids in medicinal plants using UPLC-ESI-Q-TOF MS (ultra pressure liquid chromatography-electrospray ionization-quadrupole-time-of-flight mass spectrometry). The compounds were eluted on a C18 column with 0.1 % formic acid and acetonitrile, and separated with good resolution within 13 min. Each of the separated components was characterized by precursor ions (generated by ESI-Q-TOF) and fragment ions (produced by collision-induced dissociation, CID), which were used as a reliable database. We also performed method validation: analytes showed excellent linearity ($R^2$, 0.9971-0.9996), LOD (5-25 ng/mL), LOQ (17-82 ng/mL), accuracy (91.6-97.4 %) as well as intra- and inter-day precisions (RSD, 1.8-3.2 %). In the analysis of Chelidonium majus L., magnoflorine, coptisine, sanguinarine, berberine and palmatine were detected by matching retention times and characteristic fragment ion patterns of reference standards. We also confirmed that, among the quantified components, coptisine was present in the highest quantity. Furthermore, alkaloid profiling was carried out by analyzing the fragment ion patterns corresponding to peaks of unknown components. In this manner, protopine, chelidonine, stylopine, dihydroberberine, canadine, and nitidine were tentatively identified. We also proposed the molecular structure of the fragment ions that appear in the mass spectrum. Therefore, we concluded that our suggested method for the determination of major isoquinoline alkaloids by UPLC-Q-TOF can be useful not only for quality control, but also for rapid and accurate investigation of phytochemical constituents of medicinal plants.

Anti-Helicobacter pylori Compounds from Polygonum cuspidatum

  • Khalil, Atif Ali Khan;Park, Woo Sung;Kim, Hye Jin;Akter, Kazi Marjahan;Ahn, Mi-Jeong
    • Natural Product Sciences
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    • v.22 no.3
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    • pp.220-224
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    • 2016
  • Anti-Helicobacter pylori activity guided fractionation led to the isolation of five anthraquinones, two stilbenes and one naphthoquinone from the EtOAc fraction of Polygonum cuspidatum, using silica gel column chromatography, Sephadex-LH20, MPLC and recrystallization. The chemical structures were identified to be physcion (1), emodin (2), anthraglycoside B (3), trans-resveratrol (4), anthraglycoside A (5), polydatin (6), 2-methoxy-6-acetyl-7-methyljuglone (7) and citreorosein (8) by UV, $^1H$-NMR, $^{13}C$-NMR and mass spectrometry. Anti-Helicobacter pylori activity including MIC values of each compound was evaluated. All of the isolates exhibited anti-H. pylori activity of which MIC values were lower than that of a positive control, quercetin. Compounds 2 and 7 showed potent growth inhibitory activity. Especially, a naphthoquinone, compound 7 displayed most potent antibacterial activity with $MIC_{50}$ value of $0.30{\mu}M$ and $MIC_{90}$ value of $0.39{\mu}M$. Although anti-H. pylori activity of this plant was previously reported, this is the first report on that of compounds isolated from this species. From these findings, P. cuspidatum roots or its isolates may be useful for H. pylori infection and further study is needed to elucidate mechanism of action.

Simultaneous Determination of Isoegomaketone and Perillaketone in Perilla frutescens (L.) Britton Leaves by HPLC-DAD (HPLC-DAD를 이용한 차조기 잎의 Isoegomaketone 및 Perillaketone의 동시분석법 확립)

  • Nam, Bo Mi;Lee, Seung Young;Kim, Jin-Baek;Kang, Si-Yong;Jin, Chang Hyun
    • Korean Journal of Pharmacognosy
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    • v.47 no.1
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    • pp.79-83
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    • 2016
  • This study developed an HPLC analysis method for the determination of isoegomaketone (IK) and perillaketone (PK) in Perilla frutescens (L.) Britton leaves. P. frutescens ethanol extract was optimized through an HPLC analysis using a C18 column ($250{\times}4.6mml$, D, $S-5{\mu}m$, 12 nm) with gradient elution of water and acetonitrile as the mobile phase at a flow rate of 1 mL/min and a UV detection wavelength of 254 nm. The results of this method showed linearity in the calibration curve at a coefficient of correlation ($R^2$) of IK 0.9995, PK 0.9998. The limits of detection (LOD) for IK and PK were $0.234{\mu}g/mL$ and $0.952{\mu}g/mL$. The limits of quantification (LOQ) for IK and PK were $0.017{\mu}g/mL$ and $0.043{\mu}g/mL$. The inter-day precision RSDs of IK and PK in the P. frutescens were 1.25 to 2.69% and 0.36 to 1.10%, respectively, and the intra-day precision RSDs of IK and PK were 0.96 to 2.51% and 0.90 to 1.93%, respectively. The accuracies of IK and PK were 96.31 to 97.92% and 101.26 to 105.14%. In conclusion, this method was applied successfully to the detection of IK and PK in P. frutescens.