• Title/Summary/Keyword: $MS^n$

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Purification and Partial Characterization of a Peroxidase from Perilla Callus

  • Hur, Yeon-Jae;Lee, Han-Gil;Hu, Gaosheng;Chung, Won-Bok;Jeong, Soon-Jae;Yi, Young-Byong;Nam, Jae-Sung;Chung, Young-Soo;Lee, Jai-Heon;Kim, Doh-Hoon
    • Journal of Plant Biotechnology
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    • v.34 no.4
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    • pp.355-361
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    • 2007
  • Cotyledons of perilla6 were cultured on MS medium containing 0.5 mg/l NAA and 0.5 mg/l BA for 7 weeks. The activity of perilla peroxidase was observed to increase following culture stages as assessed by peroxidase assay. A peroxidase (POD) was purified from perilla tissue cultured on MS medium for 7 weeks. The peroxidase was purified using ion exchange and gel nitration chromatography. The perilla peroxidase had a molecular mass of 30 kDa by SDS-PAGE. We showed that the N-terminal amino acid sequence of this protein shared 67% identity with the tea peroxidase. As indicated by SDS-PAGE, the banding pattern of the 30 kDa polypeptide present in total soluble protein from perilla tissue was increased following culture stages. Immunoblot analysis indicated that perilla peroxidase protein appeared after 3 weeks of perilla tissue culture, and continued to increase with extended duration of tissue culture for at least 7 weeks.

Fast and Accurate Determination of Algal Toxins in Water using Online Preconcentration and UPLC-Orbitrap Mass Spectrometry (온라인 시료주입과 UPLC-Orbitrap 질량분석법을 이용한 수질 조류독소의 고속분석방법 개발 및 환경시료적용)

  • Jang, Je-Heon;Kim, Yun-Seok;Choi, Jae-Won
    • Journal of Korean Society on Water Environment
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    • v.28 no.6
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    • pp.843-850
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    • 2012
  • Due to the fast response to algae bloom issue in drinking water treatment plant, very fast determination methodology for algal toxin is required. In this study, column switching technique based online preconcentration method was combined with high resolution full scan mass spectrometer to save sample preparation time and to obtain fast and accurate result. After parameter optimization of online preconcentration, 1mL filtered sample was directly injected to trap column with switching valve system. Next, target toxins are eluted by 98% acetonitrile and analysed with 150 - 1,100 amu scan range at 50,000 resolving power. Method detection limit (MDL) for microcystin-LR, the most toxic isomer, was 0.1 ng/mL and others such as microcystin-YR, microcystin-RR and nodularin were 0.08, 0.03 and 0.04 ng/mL, respectively. This is the best improved sensitivities with 1mL volume in the literature. Furthermore, due to the use of ultra pressure HPLC (UPLC), the whole method run was completed in 4 min. Real sample applications for 173 sample including 55 surface water and 118 treatment plant samples for raw and treated water could be done within 16 hours. In our calculation, this methodology is roughly 80% faster than the previous manual solid-phase extraction with LC-MS/MS method.

Continentalic acid, isolated from Aralia continentalis, inhibits cariogenic properties (독활로부터 분리한 continentalic acid의 항치아우식 활성)

  • Yu, Hyeon-Hee;Jeong, Seung-Il;Jung, Su-Young;Moon, Hae-Dalma;Hwang, Ji-Young;Jeong, Han-Sol;You, Yong-Ouk
    • Journal of Physiology & Pathology in Korean Medicine
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    • v.20 no.3
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    • pp.617-622
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    • 2006
  • In the present study, we investigated the anticariogenic compounds from Aralia continertalis (A. continnentalis) has been isolated and identified by MS, $^1H-NMR$ and $^{13}C-NMR$. The MeOH extract was suspended in $H_2O$ and sequentially Partitioned with $CHCl_3$, EtOAc, and n-BuOH. The $CHCl_3$ fraction showed remarkable antibacterial activity against S. mutans. The antibacterial activity compounds against 5. mutans by MIC was isolated successively through the screening system and various chromatography methods. From this active chloroform subfraction, isolation and identification finally gave continentalic acid by spectroscopic methods (MS, $^1H-NMR$ and $^{13}C-NMR$) as an active principle. The compound, continentalic acid, showed significant growth, acid production, adhesion and water-insoluble glucan synthesis inhibitory effect against 5. mutars. These results suggest that continentalic acid from A. continentalis may inhibit cariogenic properties of S. mutans and these properties may provide some scientific rationales that the local inhabitants used the extracts for treatment of dental diseases.

Phenolic Compounds from the Fruits of Paulownia coreana Uyeki (오동나무 열매 추출물의 페놀성 화합물)

  • Si, Chuan-Ling;Kim, Jin-Kyu;Kwon, Dong-Joo;Bae, Young-Soo
    • Journal of the Korean Wood Science and Technology
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    • v.34 no.1
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    • pp.79-85
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    • 2006
  • Paulownia coreana Uyeki fruits were collected, extracted with acetone-$H_2O$ (7:3, v /v), fractionated with n-hexane, methylene chloride, and ethyl acetate, then freeze dried to give some dark brown powder. The ethyl acetate soluble mixture was chromatographed on a Sephadex LH-20 column using a series of aqueous methanol and ethanol-hexane mixture as eluents. Spectrometric analysis such as NMR and MS including TLC were performed to establish the structures of the isolated compounds. From the ethyl acetate fraction, one phenolic acid (I) and five flavonoids (II~VI) and were isolated and elucidated. The antioxidative activities were tested on the isolated compounds, crude and fractionated extractives by DPPH radical scavenging method. The result showed that caffeic acid, kaempferol, luteolin, quercetin and the EtOAc soluble fraction exhibited higher activities than those of ${\alpha}$-tocopherol and BHT.

Determination of Stereochemical Structure of a Grandidentatin Isomer from Populus alba × glandulosa Bark (현사시나무 수피에서 분리한 Grandidentatin Isomer의 입체구조결정)

  • Kwon, Dong-Joo;Kim, Hyun-Seok;Lee, Phil-Ho;Bae, Young-Soo
    • Journal of the Korean Wood Science and Technology
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    • v.37 no.1
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    • pp.114-120
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    • 2009
  • The bark of Populus alba ${\times}$ glandulosa was collected, air-dried and extracted with 70% aqueous acetone. Then it was successively partitioned with n-hexane, $CH_2Cl_2$, EtOAc and $H_2O$. Repeated Sephadex LH-20 column chromatography and preparative TLC on the EtOAc soluble fraction gave a grandidentatin isomer. The structure was elucidated as grandidentatin A (cis-2-hydroxycyclohexyl 6-O-p-coumaroyl-${\beta}$-D-glucopyranoside) on the basis of spectroscopic evidences such as $^1H$-NMR, $^{13}C$-NMR, 2D-NMR and MALDI TOF-MS spectrum followed by acid hydrolysis. Grandidentatin A was identified here for the first time in Populus alba ${\times}$ glandulosa bark, and to the bset of our knowledge it has not been reported in any other literature.

Sensing of the Insecticide Carbofuran Residues by Surface Plasmon Resonance and Immunoassay (표면플라즈몬공명과 효소면역분석법을 이용한 살충제 카보후란 잔류물 검출)

  • Yang G. M.;Cho N. H.
    • Journal of Biosystems Engineering
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    • v.30 no.6 s.113
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    • pp.333-339
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    • 2005
  • The pesticide is raising public interest in the world, because it causes damage to an environmental pollution and the human health remaining agricultural products and an ecosystem, in spite of the advantages. Particularly, each country restricts the residual pesticide and induces observance about the safety and usage standard so that they can control the amount of pesticide used and defend the safety of agricultural products. The habitual practice for the analysis of the residual pesticide depends on GC (gas chromatography), HPLC (high performance liquid chromatography) and GC/MS (gas chromatography/mass spectroscopy), which triturate the fixed quantity of samples, abstract and purify as a suitable organic solvent. These methods have the highly efficient in aspects of sensitivity and accuracy. On the other hand, they need the high cost, time consuming, much effort, expensive equipment and the skillful management. Carbofuran is highly toxic by inhalation and ingestion and moderately toxic by dermal absorption. As with other carbamate compounds, it is metabolized in the liver and eventually excreted in the urine. The half-life of carbofuran on crops is about 4 days when applied to roots, and longer than 4 days if applied to the leaves. This research was conducted to develop immunoassay for detecting carbofuran residue quickly on the basis of surface plasmon resonance and to evaluate the measurement sensitivity. Gold chip used was CM5 spreaded dextran on the surface. An applied antibody to Immunoassay was GST (glutathione-s-transferase). The association and the dissociation time were 176 second and 215 second between GST and carbofuran. The total analysis time using surface plasmon resonance was 13 minutes including regeneration time, on the other hand HPLC and GC/MS was 2 hours usually. The minimum detection limit of a permissible amount for carbofuran in the country is 0.1 ppm. The immunoassay method using surface plasmon resonance was 0.002 ppm.

Fertilization and Embryo Development in Pollination and Culture for Interspecific and Intergeneric Crossing of Forage Crops (기내 수분과 배양에 의한 화본과 사료작물 종속간 수정과 배의 발육)

  • 이호진;한지연
    • KOREAN JOURNAL OF CROP SCIENCE
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    • v.32 no.4
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    • pp.455-461
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    • 1987
  • The ovaries or the ovules of grasses were pollinated and cultured in vitro to raise the interspecific or the intergeneric hybrids between tall fescue, meadow fescue, and Italian ryegrass. The isolated and suface-sterili-zed pistils were dusted with compatible pollens on stigma, on stump after removing stigma, or on excised ovule. Furthermore, the fertilized ovaries and ovules were cultured on MS, M6, or White's media and treated with plant growth regulators: IAA, kinetin, BA to promote embryo development and seed maturity. The in vitro fertilization in grass species ranged from 44 to 92% depending on ovary and pollen parents. The stigmatic pollination was resulted in 67.8% fertilization, the stump pollination 89.0%, and the excised ovule pollination 61.0%, repectively. White's medium was the most effective to provide embryo development and seed maturity in grass species. And the combined treatment of IAA 10mg/$\ell$, kinetin 0.2mg/$\ell$, was better than the non-treatment. Only two seedlings, one complete and one abnormal with root formation were obtained from 127 ovaryies cultured. The anatomy of ovules in vitro cultured was revealed the differentiation of vascular system and meristematic tissue, and the formation of sclerenchyma cells inside ovule.

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Study on the Condensation Reaction of Alkyl Glycidyl Ethers and Fatty Acids (알킬 글리시딜에테르와 지방산의 축합반응 연구)

  • Kim, Ji-Hyun;Hwang, Hyun Ah;Lee, Young-Seak;Lee, Byung Min
    • Applied Chemistry for Engineering
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    • v.23 no.4
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    • pp.416-420
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    • 2012
  • A convenient procedure for the synthesis of fatty acid diesters was studied. Long chain diesters have been used as biolubricant and transformer oils. The series of octyl, dodecyl, hexadecyl, octadecyl, and octadec-9-enyl glycidyl ether were used to synthesize those diesters. Alkyl glycidyl ethers were reacted with fatty acid such as oleic acid and octanoic acid, and octanoic acid. The one-step / two-step reactions were compared during the condensation reaction. The products were confirmed by $^{1}H-NMR$, FT-IR, and HR/MS spectra. The yield of the product 1-O-acyl-2-O, 3-O-dioctadec-9-enoylglycerol was 55~60%.

Fatty Acid Composition and Volatile Constituents of Protaetia brevitarsis Larvae

  • Yeo, Hyelim;Youn, Kumju;Kim, Minji;Yun, Eun-Young;Hwang, Jae-Sam;Jeong, Woo-Sik;Jun, Mira
    • Preventive Nutrition and Food Science
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    • v.18 no.2
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    • pp.150-156
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    • 2013
  • A total of 48 different volatile oils were identified form P. brevitarsis larvae by gas chromatography/mass spectrometry (GC/MS). Acids (48.67%) were detected as the major group in P. brevitarsis larvae comprising the largest proportion of the volatile compounds, followed by esters (19.84%), hydrocarbons (18.90%), alcohols (8.37%), miscellaneous (1.71%), aldehydes (1.35%) and terpenes (1.16%). The major volatile constituents were 9-hexadecenoic acid (16.75%), 6-octadecenoic acid (14.88%) and n-hexadecanoic acid (11.06%). The composition of fatty acid was also determined by GC analysis and 16 fatty acids were identified. The predominant fatty acids were oleic acid ($C_{18:1}$, 64.24%) followed by palmitic acid ($C_{16:0}$, 15.89%), palmitoleic acid ($C_{16:1}$, 10.43%) and linoleic acid ($C_{18:2}$, 4.69%) constituting more than 95% of total fatty acids. The distinguished characteristic of the fatty acid profile of P. brevitarsis larvae was the high proportion of unsaturated fatty acid (80.54% of total fatty acids) versus saturated fatty acids (19.46% of total fatty acids). Furthermore, small but significant amounts of linoleic, linolenic and ${\gamma}$-linolenic acids bestow P. brevitarsis larvae with considerable nutritional value. The novel findings of the present study provide a scientific basis for the comprehensive utilization of the insect as a nutritionally promising food source and a possibility for more effective utilization.

The Effects of Plant Growth Substances on the Callus Induction and Multiple Shoot Formation of Korean Lawngrass( Zoysia japonica Steud.) (식물생장조절제 처리가 들잔디의 Callus유기 및 Multiple Shoots형성에 미치는 영향)

  • 심재성;김동찬;서병기
    • Asian Journal of Turfgrass Science
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    • v.8 no.3
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    • pp.137-147
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    • 1994
  • We have established a high-frequency plant regeneration system via organogenesis from mature seed of Korean lawngrass(Zoysia japonica Steud.). The effects of 2,4-dichiorophenoxy acetic acid (2,4-D), 6-furfuryl amino purine (kinetin), $\alpha$-naphthaiene acetic acid (NAA), N6-benzyl amino pu-rine (BAP), and casein hydrolysate (CR) on cailus induction and multiple shoot formation on ex-posure to light were evaluated. Callus produced on the Murashige and Skoog (MS) medium containing 2,4-D and kinetin had high organogenesis potency. A single addition of 1.0 mg $L-^1$ 2,4-D significantly induced callus. Also, 1.0 mg L-$^1$ 2,4-D, with the addition of 0.1 mg $L-^1$ kinetin highly enlianced callus induction. The trend of cailus induction was also found on mediurn containing 0.1 mg $L-^1$ BAP with 1.0 mg $L-^1$2,4-D, and 1 g $L-^1$ CR with the addition of 1.0 mg $L-^1$ 2,4-D. However, NAA was no effective on callus formation. The growth of root was significantly high in the presence of 0.1 mg $L-^1$ kinetin compared to other concentrations. Over 2 mg $L-^1$ kinetin highly lengthened roots. Fresh weight of plantlet was highest on medium containing 0.1 mg $L-^1$ 2,4-D. Also, on medium containing 0.1 mg $L-^1$ BAP, fresh weight of piantlet was highly enhanced. BAP was significantly effective on multiple shoot formation, particularly when 2.0 mg $L-^1$ was added with 0.1 mg $L-^1$ 2,4-D. Callus induction and multiple shoot formation were achieved on MS basal medium containing 1.0 g $L-^1$ CH.

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