• 한국미생물·생명공학회지
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• 제5권1호
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• pp.36-45
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• 1977

공시균의 배양여액으로부터 독성물상을 chloroform 에 전용하여 농축한 후 avicel column chromatography 및 avicel TLC로 정제하였다. 독성물질 생성에 요구되는 영양원은 생육에 필요한 영양첨과 일치하지 않았으며 Valine, asparagine, arginine, 요소, D-glucose, D-mannose, L-rhamnose, D-ribose, D-xylose, D-arabitol, starch 등이 독성물질생성에 요구됨이 인정되었다. 또한 오탄당의 대사과정중 ribose, xylose, fructose, glucose가 대사되는 과정이 본 물질생성의 주된 경로로 추정된다. Vitamin류는 무관함에 비해 Mg가 필수적으로 요구되었다. 물질의 생성은 균의 증식에 비례되었으며 6일간 3$0^{\circ}C$에서 배양했을때 가장 많이 생성되었다. 독성물질 처리후의 잉어 조직은 아가미 신장 췌장등에서 핵진의 굴곡과 더불어 접의 심한 수축과 세포질의 괴사가 현저하였다. 본 물질의 화학식은 $C_{38}$ $H_{66}$ $NO_4$로 추정되며 UV조사시 회청색형광을 나타내었다.

• Bulletin of the Korean Chemical Society
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• 제31권12호
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• pp.3567-3572
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• 2010

Black pine barks from the southern region of Korea were extracted using pressurized hot water and the water soluble extracts were then separated in a stepwise fashion using a variety of solvents, column chromatography (CC), thin layer chromatography (TLC), and high pressure liquid chromatography (HPLC). The antioxidant activities of each fraction and the active compounds were determined based on the radical scavenging activities of 2,2-diphenyl-1-picrylhydrazyl (DPPH), reductive potential of ferric ion, and total phenol contents. A DPPH test showed that the half maximal effective concentration ($EC_{50}$ value : $6.59{\pm}0.31\;{\mu}g/mL$) of the ethyl acetate fraction (ca. 0.67%) was almost the same as that of the control compounds and inversely proportional to the value of the total phenol contents. The cell viability of the water extracts was confirmed by methyl thiazol-2-yl-2,5-diphenyl tetrazolium bromide (MTT) with enzyme linked immune sorbent assay (ELISA). Catechin, epicatechin, quercetin and ferulic acid were isolated from the ethyl acetate fraction as active compounds and identified by nuclear magnetic resonance. The antioxidant activity as value of DPPH of each of the separated compounds was lower than the ethyl acetate fraction, and ferulic acid was the lowest among these compounds.

• 대한약침학회지
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• 제15권4호
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• pp.52-65
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• 2012

Objectives: Toad venom, called Chan-Su, is a traditional Oriental medicine secreted from the auricular and the skin glands of the Bufo bufo gargarizanz Cantor or B. melanosticus Schneider and has been widely used in China, Korea and other parts of Asia for the treatment of pain, heart conditions, and cancer. We examined the concentrations of the main chemical constituents within a commercially available toad venom product and compared the levels for different extraction methods. Methods: Toad venom was extracted using either cold or hot water, ethanol (EtOH), methanol (MeOH), or ethyl acetate (EtOAc), was fractionated using precipitation or reflux, and was then analyzed using thin layer chromatography (TLC), high-performance liquid chromatography (HTLC), and liquid chroma-tography - mass spectrometry (LC-MS). Individual components were identified by comparisons of the retention times, the ultraviolet spectra, and mass spectras and differences in chemical constituents for different solvents and extraction methods are presented. Results: Components with authentic standards, including serotonin and bufodienolides (cinobufagen, bufalin, cinobufalin, and resibufogenin), were detected. The water extract of toad venom contained the greatest amount of serotonin ($75.7{\pm}0.1$ mg/g), but very small amounts of bufodienolides ($3.8{\pm}0.0$ mg/g). In contrast, the use of MeOH or EtOH extraction solutions resulted in 5-26 times higher concentrations of bufodienolides, with only trace amounts of serotonin. The relative and the absolute concentrations of the component also varied based on the extraction method; i.e., EtOH extracts yielded the greatest total amounts of bufodienolides, and EtOAc precipitation had the lowest amounts of bufodienolides. Conclusions: Toad venom consists of serotonin and several bufodienolides, and the choice of solvent to extract chemical the constituents is important as a way to enrich the purported active components for treating different conditions.

• 한국수산과학회지
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• 제15권1호
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• pp.59-73
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• 1982

두족류의 근육지질의 특징을 알기 위하여 3종의 두족류를 사료로 하여 이들 근육에서 총지질을 적출하였고, 이를 thin layer chromatography, column chromatography 및 gas-liquid chromatography 그리고 TLC-scanner를 이용하여 총지질을 구성하는 지질성분. 지방산조성, sterol조성 그리고 총지질에서 분획한 중성지질, 당지질 및 린지질의 지방산조성을 정량하여 다음과 같은 결과를 얻었다. 1) 각 시료의 총후무사양은 생시료에 대하여 문어가 $0.5\%$, 낙지가 $0.8\%$ 그리고 꼴뚜기가 $0.6\%$였고 이들 평균지질함양은 $0.6\%$였다. 그리고 총지질의 전지방산양은 문어가 $19.3\%$, 낙지가 $47.8\%$,꼴뚜기가 $38.4\%$였으며, 불광화물양은 문어가 $10.9\%$, 낙지가 $15.8\%$, 꼴뚜기가 $41.1\%$로서 높은 함양을 나타냈다. 2) 총지질을 구성하는 성분으로는, sterol성분과 극성지질일색소 혼합층의 함양분포가 총지질에 대하여 각각 $27.0\sim35.5\%$ 그리고 $38.3\sim63.4\%$로서 시료에서 다 같이 높은 함양을 나타내고 있어 총지질이 주성분을 이루며, 문어의 근육에는 다른 시료보다 sterol성분의 함양이 높았다. 그리고 그가 소양성분으로 triglyceride, 유리지방산 및 탄화수소-esterified sterol혼합층이 분리 동정되었다. 3) 총지질의 지방산조성은 시료에서 다 같이 Palmitic acid가 주성분을 이루고 그외 oleic acid, linoleic acid, octadecatetraenoic acid 및 eicosapentaenoic acid가 구성지방기의 주체를 이루고 있다. 특히 문어의 총지질에는 palmitic acid와 eicosapentaenoic acid가 서로 비슷한 양으로 존재하면서 주성분을 이루고 있었다. 4) sterol조성에 있어서는 시료에서 다 같이 전sterol성분에 대한 cholesterol의 함양분포가 $82.4\sim89.1\%$로서 가장 많았고 그외 소양성분으로서 22-dehydrocholesterol 과 24-methylenecholesterol이 각 시료에 공통적으로 함유하고 있었다. 5) 총지질에서 분획한 비극성복질(신성지질)과 극성지질의 함양비는 시료에서 다 같이 비극성낙질보다 극성지질의 함양이 훨씬 높았다. 그리고 극성지질중에서도 당지질보다 린지질의 함양이 월등히 높았다. 6) 중성지질을 구성하는 성분으로는 유리 sterols의 함양분포가 중성서질에 대하여 $50.0\sim70.5\%$로서 주성분을 이루며, 그 다음 서로 비슷한 양으로 나타나는 esterified sterol, 유리지방산 및 triglyceride가 분리 동정되었다. 그리고 중성지질을 구성하는 지방산은 palmitic acid, oleic acid, hexadecadienoic acid 주성분이었다. 7) 당지질을 구성하는 지방산은 palmitic acid가 가장 많고 그가 oleic acid, linoleic acid, octadecatetraenoic acid, myristic acid가 구성지방산의주함분을 이룬다. 그러나 특히 꼴뚜기의 당지질에는 이들 지방산외에 arachidonic acid의 함양이 월등히 높아 주성분의 하나였다. 8) 린지질을 구성하는 지방산은 palmitic acid함양이 가장 많았고 그가 oleic acid, linoleic acid, octadecatetraenoic acid가 구성지방산의 주성분이었다. 본 연구는 1981년도 문교부연구조성비의 지원으로 이루어 졌음을 밝힌다.

• 한국식품저장유통학회지
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• 제10권4호
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• pp.527-533
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• 2003

호박씨는 단백질과 지방의 함량이 높고 영양가치가 높아 식품가공 재료로서 널리 이용될 수 있으나 그 동안 과자, 스넥 제품 등의 단순가공 소재로서 이용되어 왔다. 그러므로 호박씨의 소비를 증진시키고 새로운 기능성 식품을 개발하기 위하여 호박씨를 발아 성장시키면서 각 부위의 일반영양성분, 지방산, 아미노산, L-ascorbic acid, $\beta$-carotene의 함량변화를 측정하여 영양학적 가치를 평가하였으며, 발아 성장과정 중 생성되는 고미성분의 구조를 동정하기 위하여 각종 용매 순차 분획, thin layer chromatography, HPLC의 분리과정을 거쳐서 정제한 고미물질을 mass spectrum, $^1$H-NMR spectrum, $^{13}$C-NMR spectrum을 이용하여 동정하였다. 호박씨나물의 중량은 발아 8일에 348.4% 증가하였으며 뿌리와 줄기의 길이는 8일째까지 급격히 증가하였다. 일반영양성분은 머리부분과 줄기부분 모두 단백질과 지질의 함량은 감소하였고 섬유소, 회분, 가용성 무질소물의 함량은 증가하였으며, linoleic acid, oleic acid, palmitic acid, stearic acid가 주요지방산으로 나타났고, palmitic acid는 증가하는 반면 linoleic acid는 점점 감소하는 경향을 보였다. 호박씨나물의 머리부분의 구성아미노산 glycine, alanine, arginine, cystein proline의 순으로 많았으며, 유리아미노산은 arginine, threonine, alanine, glutamine의 순으로 많았다. L-ascorbic acid와 $\beta$-carotene은 발아 성장하면서 점점 증가하는 경향을 보였으며, 머리부분에 생성되는 고미성분은 cucurbitacin glycoside로 판명되었으며, 호박씨에서는 고미성분이 검출되지 않았으나 발아 8일째 머리부분의 고미성분은 42.2mg/kg이 함유되어 있다.

• 한국연초학회지
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• 제27권1호
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• pp.11-18
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• 2005

The nicotine converter genotypes of burley tobacco (Nicotiana tabacum L.), which convert nicotine to nornicotine, contain a high amount of nornicotine that degrades tobacco quality and smoking taste. Elimination of nicotine converter plants before seed harvesting is required for breeding nicotine low-converter lines and for increasing their seed production. This study aims to develop a rapid and convenient method of identifying nicotine converter plants of burley breeding lines of KB9118(KB108) using thin-layer chromatography (TLC) and isatin coloration method. Out of 223 plants in 10 lines harvested at maturity in 2002, 102 plants ($45\%$) were identified as nicotine converters by TLC of tobacco leaves air-cured. For 16 lines selected as low-converters in 2002, 148 plants grown in the field in 2003 were tested by the isatin coloration method using two detached leaves at the flowering stage thoroughly sprayed with $1\%\;NaHCO_3$ solution and cured in conditioned chambers for the early identification of nicotine to nornicotine conversion. From these samples, 46 plants ($31\%$) in 4 lines were identified as nicotine converters, indicating that the ratio of converters significantly decreased by one time selection. Mean percent conversion of non-screened lines was $14\%$ higher than that of following generation. Therefore in the burley tobacco, a rapid and convenient means of identifying and removing nornicotine converter plants by the isatin coloration method during growth in the greenhouse or field were effective in reducing the converter plants in the following generation.

• Journal of Ginseng Research
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• 제23권1호
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• pp.55-59
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• 1999

For the first time we have developed a simple method for conjugation between ginsenoside Rf (Rf) and bovine serum albumin (BSA) or ovalbumin (OVA) by a periodate oxidation method. The Rf-BSA conjugate is confirmed by ultraviolate (UV) scanning, thin layer chromatography (TLC), and SDS-gel electrophoresis. In UV scanning Rf showed three small peaks approximately at 230, 265 and 280 nm. BSA showed a peak at 280 nm. The Rf-BSA conjugate showed right shifted three peaks that BSA alone did not show. In TLC analysis, the Rf-BSA conjugate did not show mobility in silica gel but showed a slight stream of trace. In SDS-gel electrophoresis, Rf-BSA conjugate show a slight less mobility than BSA alone. Rf-OVA conjugate also showed similar patterns with Rf-BSA conjugate. These results demonstrated that periodate oxidation method could be used to produce a stable Rf-BSA or Rf-OV A conjugate and also could be applied for other ginsenoside(s).

• Asian Pacific Journal of Cancer Prevention
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• 제16권16호
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• pp.7337-7342
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• 2015

The study aimed to investigate the profile of alkaloids in two ethyl acetate soluble fractions, namely fractions A and B from an ethanolic extract of Ficus septica leaves and cytotoxic effect on T47D breast cancer cells. Preparation of both fractions involved maceration of leaves with 70% (v/v) ethanol, filtration with $Al_2O_3$, precipitation with 0.1 N HCl, Mayer reagent, and 0.1 N NaOH, and also partition with ethyl acetate. Qualitative thin layer chromatography (TLC) was conducted to determine the profile of alkaloids in the two fractions, using alkaloid specific reagents such as Dragendorff, sodium nitrite, and Van Urk-Salkowski. Cytotoxic effects of both fractions on T47D cells were evaluated using MTT assay with a concentration series of 1.56; 3.12; 6.25; 12.5; 25 and $50{\mu}g/mL$. The TLC test showed that fractions A and B contained alkaloids with Rx values of 0.74 and 0.80 for fraction A and 0.74, 0.84, 0.92 for fraction B with regard to yohimbine using the mobile phase of n-buthanol:glacial acetic acid:distilled water (3:1:1 v/v/v). Moreover, an indole alkaloid was detected with Rx values of 0.80 and 0.84, respectively. Fractions A and B exhibited high cytotoxic effects on T47D cells with IC50 values of 2.57 and $2.73{\mu}g/mL$, respectively. In conclusion, overall the results of this study showed that fractions of Ficus septica contain alkaloids including indole alkaloid or its derivatives and possess a cytotoxic effect on T47D cells. This research supports the idea that alkaloids in F. septica have anticancer activity.

• 한국산림과학회지
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• 제81권2호
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• pp.183-190
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• 1992

기내배양된 현사시나무의 조직배양세포로부터 안토시아닌 생성과 세포생장에 적합한 요인을 구명하여 차후 기내배양을 통한 2차대사산물생산의 기초자료를 제공하고자 연구를 수행하였다. Callus는 0.5mg/l 2, 4-D, 0.1mg/l BAP을 첨가한 MS배지에서 유발시켜 같은 배지에서 증식하였다. 안토지아닌 생산은 MS 기본배지를 기준으로 하여 질산염은 12.5% 감량시키고 인산염은 400%로 증가시켜 5% sucrose와 1.0mg/l IAA 및 1.0mg/l BAP를 첨가한후 7,000 lux의 연속광 하에서 배양했을 때 가장 높게 나타났다. 그러나 세포의 생장은 MS 기본배지를 기준으로 하여 질산염은 50% 감량시키고 인산염은 400%로 증가시켜 5% sucrose와 0.5mg/l 2, 4-D를 첨가한 배지에서 가장 양호한 결과를 얻었다. 안토시아닌의 동정은 1% 염산-메탄올의 유기용매로 추출하여 정제 후 TLC와 UV spectrophotometer로 확인한 결과 pelargonidin 3-rhamnoside-5-glucose로 추정되었다.

• Reproductive and Developmental Biology
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• 제35권3호
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• pp.265-271
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• 2011

This study was undertaken to find out the effect of methyl-beta-cyclodextrin (MBCD) on cold shock and membrane cholesterol quantity of sperm during the freezing process in miniature pigs. For this study, semen ejaculated from PWG M-type miniature pig was diluted that freezing solution (with egg yolk group) and m-Modena B (without egg yolk group) treated with 0, 1, 5, 10 and 20 mM MBCD before freezing process. The diluted semen was monitored sperm ability at room temperature, after cooled until $5^{\circ}C$ and after forzen-thawed for cold shock test of spermatozoa. Also, membrane cholesterol of sperm was extracted by folch solution at the same time sperm ability was assessed for viability and acrosomal status. The membrane cholesterol quantity was measured by thin-layer chromatography (TLC) method. The result, viability and acrosome integrity in semen diluted without egg yolk groups were decreased at all temperature range by increasing of MBCD concentration. In particular, sperm of egg yolk group was showed that significantly higher viability and lower acrosome damage when treated with 5 mM MBCD (p<0.05). The results of TLC experiment, cholesterol amounts were increased with MBCD cocentration in egg yolk, and decreased with MBCD concentration in m-Modena B. In cryopreservation efficiency, there was no significant difference at viability, and acrosomal state of sperm in 5 mM MBCD concentration was significantly lower in acrosome damage than other groups (p<0.05). Therefore, the addition MBCD in egg yolk was protected spermatozoa from cold shock injury. This protective effect of MBCD may be due to addition of sperm membrane cholesterol.