• 한국식품영양과학회지
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• 제21권1호
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• pp.76-83
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• 1992

장독성대장균은 영유아설사 및 여행자설사의 주요한 원인균의 하나로서 이들이 생산하는 내열성장독소는 설사 유발 원인물질로 이들 생산균주는 비병원성 대장균과 비교하여 검정할 수 있다. 본 실험에서는 이러한 내열성장독소를 순수정제하였다. 저자들이 보고한 바 있는 형질전환균주 eKT-53을 사용하여 10L의 succinate salts 배지에 배양한 배양액을 30,000g로 원심분리하여 조 ST 용액을 사용하여 몇 단계의 정제과정을 통하여 정제하였다. 즉 Amberlite XAD-2 chromatography, DEAE-Sephacel anion exchange chromatography, Bio-Gel P-6 get filtration, Polyacrylamide slab gel 전기영동을 통하여 정제하고 장독소의 정제도 및 장독소의 생물학적 최소단위량을 조사하였다. 최종적으로 정제되어진 ST는 조 ST 용액에 비해 113배 더 정제되었고 회수율은 11%였다. 또한 생물학적 최소단위량이 2.8ng이였고 SDS-polyacrylamide disc gel상에서 단일 band를 나타내어 거의 순수정제된 것으로 보이며 면역을 위한 항원으로 이용할 수 있을 것으로 생각된다.

• 한국약용작물학회지
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• 제12권3호
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• pp.203-208
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• 2004

참돌꽃의 캘러스는 0.5 mg/l NAA와 1 mg/l BA가 조합 첨가된 1/2MS 배지에서 유도하였으며, 현탁세포배양은 0.5 mg/l NAA와 1 mg/l BA가 조합 첨가된 2B5 배지에서 확립되었다. 한편, 참돌꽃 뿌리로부터 반복된 silicagel colum chromatography와 분취 HPLC를 통하여 순수한 salidroside 결정을 얻었으며, 얻어진 결정체는 $^1H-NMR,\;^{13}C-NMR$ 및 $^1H-^1H$ COSY 등의 분광학적 분석으로 구조를 동정 확인하였다. 확립된 세포배양계에서 salidroside 함량은 원뿌리 0.17%에 비교하여 광배양(0.12%) 암배양 (0.286%) 및 sucrose 5%에서 함량이 0.41% 까지의 함량변화를 나타내었다.

• The Plant Pathology Journal
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• 제15권1호
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• pp.14-20
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• 1999

Twenty isolates of Helminthosporium species were obtained from various grass plants and tested for controlling efficacy on the development of plant diseases. An isolate of Helminthosporium sp. TP-4 was chosen and six antibiotic substances were purified from cultures of the fungus by repeated silica gel column chromatography and preparative thin-layer chromatography. They were identified as ophiobolin a, 6-epiophiobolin A, 3-anhydroophiobolin A, 3-anhydro-6-epiophiobolin A, iphiobolin B, and iphiobolin I mainly by mass spectrometry and nuclear magnetic resonance spectrometry. Ophiobolins inhibited the growth of a grampositive bacterium Streptomyces griseus, but were not active against gram-negative bacteria. They also showed an antifungal activity. In in vivo tests, iphiobolin B exhibited potent controlling activities against rice blast, tomato late blight, and wheat leaf rust with control values more than 90% and 70% at concentration of $500\mu\textrm{m}$/ml and 100 ${\mu}{\textrm}{m}$/ml. Ophiobolin A and 6-epiophiobolin A controlled the development of wheat leaf rust more than 80% at concentrations of 100 /ml and $500\mu\textrm{m}$/ml respectively. 3-Anhydro-6-epiophiobolin A was not active against any plant disease. On the other hand, the A-series ophiobolins other than 3-anhydroophiobolin A showed stronger phytotoxic activity in a leaf-wounding assay using 8 plant species than those of 3-anhydroophiobolin A, ophiobolin B, and ophiobolin I. The results indicate that there is little correlation between antifungal activity and phytotoxicity of ophiobolins.

• 생약학회지
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• 제41권2호
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• pp.73-88
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• 2010

Traditionally, the root of Lithospermum erythrorhizon Sieb. et Zucc(L.E) has been used as efficacious therapy for inflammation, burns, frostbite and skin ailments (e.g eczema and psoriasis). It contains isohexenylnaphthoquinone derivatives (shikonin and its esters) and furylhydroquinones (shikonofurans) in lipophilic fractions and caffeic acid oligomers (rosmarinic acid, lithospermic acid B) in polar fractions. Recently, new preparative isolation and analysis procedures of shikonin along with its oligomers from the extract of L. erythrorhizon by the combination of high-speed counter-current chromatography with high-performance liquid chromatography-diode array detection have also been introduced. Although there have been many reports on the wound healing, antiinflammatory, and anticancer effects, the research on the effects of anti-atopic dermatitis of the root of L. erythrorhizon were relatively scarce. However, in recent years, new information gathered from research efforts, on the anti-atopic dermatitis properties of the extract or constituents of L. erythrorhizon has been accumulated. In this paper, the findings and advance on the in vitro and in vivo activities of L. erythrorhizon and its constituents especially focused on antiinflammatory and anti-atopic dermatitis effects are summarized. The phytochemical constituents of L. erythrorhizon or its tissue cultures are also presented. Although there are few to verify or refute its activity in human, one result of clinical study of the extract of L. erythrorhizon on the atopic dermatitis patients was introduced to assess the possibility of its clinical use. The reported mechanisms of action and in vivo pharmacological studies in different animal models for the various types of extracts or constituents of L. erythrorhizon are supportive of its therapeutic potential or dietary supplement, however, more evidence from clinically relevant models, as well as systemic studies on the active constituents or the various types of standardized extracts at the cellular and molecular level, are required.

• Applied Biological Chemistry
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• 제46권1호
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• pp.50-54
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• 2003

호장근에서 분리한 emodin은 VEGF로 유도된 혈관신생의 한 단계인 혈관내피세포의 이동을 강하게 억제하였다. 또한 emodin은 혈관내피세포 이동을 억제시킨 $0.1\;{\mu}g/ml$ 농도에서 시험관내 혈관신생을 억제하였으며, 그 효과는 농도의존적인 양상을 보였다. 생체내 혈관신생 모델인 CAM assay에서도 emodin은 혈관신생을 억제하였다. 이러한 결과는 emodin이 현재까지 보고된 여러 가지 생리활성 이외에 혈관신생 억제활성을 가지고 있다는 사실을 보여주는 것이다. 따라서 emodin을 함유하고 있는 호장근은 혈관신생 관련 질환에 대한 천연물 유래 치료제의 개발을 위한 중요한 식물자원으로 활용될 수 있을 것이다.

• 한국수산과학회지
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• 제34권5호
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• pp.556-562
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• 2001

본 연구에서는 비교적 성장이 빠르고 배양이 용이한 황갈편조 식물문 중 규조강에 속하는 P. tricornutum으로부터 선정하여 항 산화성 물질을 분리$\cdot$정제하여 물질의 구조를 밝혔다. P. tricornutum의 각 유기용매 추출물의 항산화성은 chloroform획분이 가장 좋았으며, 이 획분을 시료로 하여 silica gel column chromatography, PTLC 및 HPLC를 사용하여 항산화성 물질을 분리$\cdot$정제하였다. Silica gel column을 사용하여 분리된 획분의 항산화성은 dichloromethane: methanol (5: 1)로 용출시킨 획분에서 가장 높았으며, 이 획분의 PTLC 획분은 항산화성이 $20\%$ 증가하였다. 최종 정제된 물질의 $IC_50$값은 $8.3{\mu}g/mL$으로 화학적으로 합성된 항산화제인 BHT 및 BHA보다는 낮았지만 천연 항산화제인 $\alpha-tocopherol$보다는 다소 높게 나타났으며, 이 물질의 화학적 구조는 carotenoid류인 zeaxanthin으로 동정되었다.

• Preventive Nutrition and Food Science
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• 제18권1호
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• pp.76-79
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• 2013

We evaluated antioxidant activities of heated pear juice (HPJ) exposed to 120, 130, and $140^{\circ}C$ for 2 hr. HPJ was partitioned using n-hexane, chloroform, ethyl acetate, n-butanol, and water. The ethyl acetate fraction treated at $130^{\circ}C$ for 2 hr showed strong antioxidant activity; thus, this extract was isolated and purified using silica gel column chromatography and preparative high performance liquid chromatography. The structure of the purified compound was determined using ultraviolet and mass spectrometry, $^1H$-nucelar magnetic resonance (NMR), and $^{13}C$-NMR. Antioxidant activities of the isolated compound were evaluated and compared with ${\alpha}$-tocopherol, ascorbic acid, and butylated hydroxytoluene (BHT) using DPPH and ABTS assays. The isolated compound was identified as 2,3-dihydro-3,5-dihydroxy-6-methyl-4H-pyran-4-one (DDMP). The DPPH radical-scavenging activity ($IC_{50}$) of DDMP occurred in the following order: ascorbic acid ($45.3{\mu}g/mL$) > ${\alpha}$-tocopherol ($69.2{\mu}g/mL$) > DDMP ($241.6{\mu}g/mL$) > BHT ($268.0{\mu}g/mL$). Furthermore, DDMP showed strong ABTS radical-scavenging activity (569.0 mg AA eq/g).

• Journal of Ginseng Research
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• 제35권4호
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• pp.487-496
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• 2011

Good manufacturing practice (GMP)-based quality control is an integral component of the common technical document, a formal documentation process for applying a marketing authorization holder to those countries where ginseng is classified as a medicine. In addition, authentication of the physico-chemical properties of ginsenoside reference materials, and qualitative and quantitative batch analytical data based on validated analytical procedures are prerequisites for certifying GMP. Therefore, the aim of this study was to propose an authentication process for isolated ginsenosides $Rb_1$ and $Rg_1$ as reference materials (RM) and for these compounds to be designated as RMs for ginseng preparations throughout the world. Ginsenoside $Rb_1$ and $Rg_1$ were isolated by Diaion HP-20 adsorption chromatography, silica gel flash chromatography, recrystallization, and preparative HPLC. HPLC fractions corresponding to those two ginsenosides were recrystallized in appropriate solvents for the analysis of physico-chemical properties. Documentation of the isolated ginsenosides was made according to the method proposed by Gaedcke and Steinhoff. The ginsenosides were subjected to analyses of their general characteristics, identification, purity, content quantitation, and mass balance tests. The isolated ginsenosides were proven to be a single compound when analyzed by three different HPLC systems. Also, the water content was found to be 0.940% for $Rb_1$ and 0.485% for $Rg_1$, meaning that the net mass balance for ginsenoside $Rb_1$ and $Rg_1$ were 99.060% and 99.515%, respectively. From these results, we could assess and propose a full spectrum of physicochemical properties for the ginsenosides $Rb_1$ and $Rg_1$ as standard reference materials for GMP-based quality control.

• Applied Biological Chemistry
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• 제39권4호
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• pp.320-326
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• 1996

우리나라에서 식용으로 이용되는 진달래꽃으로부터 7성분의 항산화성이 있는 플라보노이드 화합물이 분리 동정되었다. 이들 화합물의 구조는 IR, UV, FAB-MS, $^1H\;NMR$ 및 $^{13}C\;NMR$에 의해서 얻어진 분광학적인 결과에 근거하여 afzelin, ampelopsin, catechin, myricetin, myricitrin, quercetin 및 quercitrin인 것으로 밝혀졌다. 이들 화합물은 2개의 flavonol, 3개의 flavonol glycoside, 1개의 flavane 및 1개의 dihydroflavonol로 이루어졌다. 에틸에테르 및 초산에틸 구분에 존재하는 flavonol glycoside (14.4 g)는 polyamide C-200 관 크로마토피법, 분취용 박층크로마토그래피법, 재결정화법, sephadex LH-20 관 크로마토그래피법을 통해서 양쪽 구분으로부터 최종적으로 회수된 총 flavonoid량 (17.6g의 82%에 달하였다. 항산화성은 티오시안산철의 존재하에서 리놀레산의 에타놀용액 중에서 측정되었다. 항산화 효능성은 afzelin<$\alpha-tocopherol$

• Food Science and Biotechnology
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• 제16권6호
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• pp.963-966
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• 2007

The objectives of this study were to identity antioxidant substance in heated garlic juice (HGJ). We evaluated the antioxidant activities of heated garlic juice exposed to 120, 130, and $140^{\circ}C$ for 2 hr. The HGJ was partitioned using the solvents of hexane, chloroform, ethyl acetate, butanol, and water. The ethyl acetate fraction of HGJ treated at $130^{\circ}C$ for 2 hr showed strong antioxidant activity; this extract was isolated and purified using silica gel column chromatography and semi-preparative high-performance liquid chromatography. The structure of the purified compound was determined using spectroscopic methods, i.e., ultraviolet, mass spectrometry, infrared, $^1H$ NMR, $^{13}C$ NMR, DEPT, HMBC, and HMQC. The isolated compound was identified as thiacremonone (2,4-dihydroxy-2,5-dimethyl-thiophene-3-one). Thiacremonone showed strong 1,1-diphenyl-2-picrylhydrazyl (DPPH) radical-scavenging activity, with a 50% inhibition concentration ($IC_{50}$) of $22.25{\pm}0.44\;{\mu}g/mL$, which is much higher than that of the antioxidants ascorbic acid ($30.06{\pm}0.42\;{\mu}g/mL$), ${\alpha}$-tocopherol ($71.30{\pm}0.97\;{\mu}g/mL$), and butylated hydroxyanisole ($50.54{\pm}0.94\;{\mu}g/mL$).