• Title/Summary/Keyword: thin-layer chromatography

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Improved Phosphotyrosine Analysis by TLC and HPLC

  • Song, Young-Me;Yoo, Gyurng-Soo;Lee, Seung-Ki;Choi, Jung-Kap
    • Archives of Pharmacal Research
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    • v.16 no.2
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    • pp.99-103
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    • 1993
  • We describe here the conditions of thin layer chromatography (TLC) and high pressures liquid chromatography (HPLC) to improve the analytical method of phosphotyrosine (p-Tyr) in biological sample. TLC was performed on silica plate with the mixture of propanol and water (2.1 : 1 v/v) as a mobile phase and $R_1$ values were 0.42, 0.39 and 0.33 for phosphotyrosine, phosphothreonine and phosphoserine, respectively. HPLC was performed on $NH_2$ column with a mobile phase of potassium biphosphate solution by UV deterction at 192 nm. The optimum condition of HPLC was obtained at 0.01 M, pH 4.5 with a clear separation within 12 min. These procedures have been applied to the analysis of phosphotyrosine obtained from tyrosine-phosphorylated enolase. Both TLC and HPLC methods were suitable to analyze tyrosine-phosphorylated protein without being affected by contaminants from hydrolysates.

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Chlorination of Phenyl Derivatives : Chlorination of ethyl -${\alpha},{\beta}-dichloro-{\beta}$-phenyl propionate under gamma ray irradiation (芳香族 誘導體의 염素化反應 Ethyl-${\alpha},{\beta}-dichloro-{\beta}$-phenyl propionate의 gamma 線 鹽素化反應)

  • Kim, You-Sun;Kim, Ki-Soo
    • Journal of the Korean Chemical Society
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    • v.12 no.2
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    • pp.55-60
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    • 1968
  • Chlorination of aromatic derivatives under UV light and ${\gamma}$-ray irradiation was studied. Ethyl ${\alpha} ,{\beta} -dichloro-{\beta}$-phenyl propionate gave the p-chlorophenyl derivatives when chlorination was done under UV light. The same type of the product was obtained in the reaction where the mole ratio of the ester and chlorine was 1 to 2 and the chlorination was done under ${\gamma}$-ray irradiation. When the mole ratio of the ester and chlorine was 1 to 8, the chlorination reaction under ${\gamma}$-ray irradiation gave a poly-chlorinated derivatives which was identified as a side chain chlorinated p-chlorophenyl derivatives. Ethyl ${\alpha} ,{\beta} -dichloro-{\beta}$-(p-chlorophenyl) propionate gave the same type of the side chain chlorinated p-chloro derivatives by the chlorination under ${\gamma}$-ray irradiation, whereas ethyl ${\alpha} ,{\beta} -dichloro-{\beta}$-(o-chlorophenyl) derivatives gave o,p-dichlorophenyl derivatives. The identifications of the products were based on a radio thin layer chromatography and activation analysis of chlorine contents of product. The chlorination reaction was discussed in regards to the effect of phenyl substituents to the formation of reaction product and the procedures were described.

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Studies on the Constituents of Spiraea Koreana Nakai (참조팝나무의 成分 Alkaloid 에 關한 硏究)

  • Jin, Kab-Dukc
    • Journal of the Korean Chemical Society
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    • v.11 no.3
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    • pp.111-116
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    • 1967
  • A new alkaloid named Spirajine(m.p. 182~$184^{\circ}C$ $[{\alpha}]d^{19}+3.4^{\circ}$ in $CHCl_3$, $C_{23}H_{33}NO_3$, colorless prism) was isolated from the leaves of Spiraea Koreana Nakai (Spiraeceae) (Korean name "Chamjopab namu") which grows in the mountaineous area of Korea, by process of Scheme I (yields 0.13%). Another two unidentified alkaloids (not yet crystallized) were separated by the method of thin layer chromatography. (The Rf values of the two unidentified alkaloids were 0.66, 0.77, respectively and Spirajine 0.72) Spirajine were subjected to the structural investigation with the use of ultra violet and infra red spectrophotometry, and opical rotatory dispersion. The alkaloid contains two ketonic carbonyl groups, tertiary hydroxyl group, methyl groups, N-methyl group and both cyclohexane ring and cyclopentane ring.

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Development of a Thin Layer Chromatography Experimental Kit for Middle School Students (중학생들을 위한 얇은막 크로마토그래피 실험 키트의 개발)

  • Jae Jeong Ryoo;Won-Kyu Park
    • Journal of the Korean Chemical Society
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    • v.47 no.4
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    • pp.401-411
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    • 2003
  • One of the most important characteristics in the seventh educational course is an enforcement of differentiated level curriculum. Therefore, development of various different level educational subjects is needed. A new chromatographic experimental kit which is helpful for middle school students to understand chromatographic principles is needed to correct some mistakes of chromatographic part within the 6th educational course's middle school textbook. In this research, a new thin layer chromatography(TLC) experimental kit for middle school students are developed by using acid-base indicator as samples and ethyl alcohol, ethylacetate, hexane as eluents. The kit is applied to two kinds of middle school student groups(general group & genius group) and two kinds of teacher groups(science teacher & non-science teacher) and an efficacy of the new chromatographic kits are evaluated through making up some questions.

Clean Analysis Not to Use Harmful Solvent for TLC Identification Test of the Herbal Medicine (생약의 확인시험법에 대한 유해용매 대체시험법 설정)

  • Kim, Ji-Yeon;Park, Ju-Young;Shim, Young-Hoon;Cho, Chang-Hee;Lee, Jong-Hwa;Lee, Dong-Jin;Kim, Bo-Joon;Kim, Dong-Sup;Seong, Rack-Seon;Kim, Jong-Hwan;Lim, Sook;Shin, Jin-Seon;Jang, Seung-Yeup
    • Korean Journal of Pharmacognosy
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    • v.41 no.2
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    • pp.136-140
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    • 2010
  • This study was carried out to utilize non-toxic solvents instead of harmful solvents for the TLC(Thin Layer Chromatography) identification test of herbal medicines. It is recommended not to use harmful solvent such as chloroform at a viewpoint of clean analysis. In this study, we revised the identification test of 10 items in the Korea Pharmacopoeia(KP) and the Korean Herbal Pharmacopoeia(KHP) such as Cornus officinalis S. containing the harmful solvents on the developing solvent and established identification test that is utilized non-toxic solvents by HPTLC(High Performance Thin Layer Chromatography).

Physico-chemical Characteristics of the Inhibitory Substance on the Germination of Mung Bean (녹두 발아 조해물질(阻害物質)의 이화학적(理化學的) 성질(性質))

  • Kim, Kwang-Hyeon
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.14 no.1
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    • pp.57-60
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    • 1985
  • The inhibitory substance on the growth of mung bean seedling was amorphous crystal that had a single spot on silica gel thin layer chromatography under ultra-violet irradiation in dark room. The Rf value was 0.85 in n-butanol-acetic acid-formic acid-water(15 : 12 : 3 : 10) system, and 0.92 in n-butanol-acetic acid-water(4 : 1 : 5) system, respectively. Biochemical reaction of the inhibitor was negative to $FeCl_3$, pyrimidine, ninhydrine, folin, fehling, Salkowski and Ehrlich reaction, but the inhibitor had a blue fluorescence from irradiation of ultra-violet rays and had maximum absorption at 252nm on UV spectrum. On IR spectrum, the Peaks of the inhibitor appeared at $3300{\sim}3500,\;2900{\sim}3000,\;1600{\sim}1700,\;and\;1400cm^{(-1)}$. The strain tested was identified to be similar Streptomyces iuteogriseus by morphorogical and physiological characteristics.

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Studies on the Change of Sugars in Sweet Potatoes on Heating (가열(加熱)에 의한 고구마의 당(糖)의 변화(變化))

  • Rhee, Eun-Hee;Ahn, Seung-Yo
    • Applied Biological Chemistry
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    • v.24 no.4
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    • pp.245-251
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    • 1981
  • A study was conducted on the conversion of starch in sweet potatoes to sugar by the amylolytic enzymes during baking. Sugars were extracted with ethanol from the raw and baked sweet potatoes at the temperature of $55{\sim}57$, $70{\sim}75$, and $90{\sim}95^{\circ}C$. The individual sugars in the extracts was identified by thin-layer chromatography- and the individual sugar content was determined by high -performance liquid chromatographic analysis. Sugars identified from the raw and baked sweet potatoes at the temperature of $55{\sim}57^{\circ}C$ are glucose, fructose, and sucrose. Sucrose, maltose, glucose, and fructose were identified is the baked sweet potatoes at tile temperature of $70{\sim}75$ and $90{\sim}95^{\circ}C$. There was no significant increase in glucose, fructose and sucrose content during baking. Maltose was formed only above the gelatinization temperature.

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Studies on the Production of Gluconic Acid by Resting Cell System of Aspergillus niger (Aspergillus niger의 휴지균체에 의한 Gluconic Acid생성에 관한 연구)

  • 정지관;양호석;신규철;양한철
    • Microbiology and Biotechnology Letters
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    • v.9 no.1
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    • pp.7-19
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    • 1981
  • The production of gluconic acid from glucose by the resting cell system of Aspergillus niger was studied. It was found that the conversion products from glucose by the resting cell system were markedly influenced by the pH, temperature, substrate concentration, aeration, metal ions, cultivation time and storage conditions of the resting cells. Conversion products were identified as gluconic acid by the thin layer chromatography and infrared spectrophotometry. These conversions were greatly stimulated by addition of $Mg^{++}$, and S $n^{++}$, but showed inhibitory effects by C $u^{++}$, H $g^{++}$, C $d^{++}$, A $g^{+}$ and cyanide. For the optimum cell storage, it was effective to be kept at -$25^{\circ}C$ in 0.05M phosphate buffer solution of pH 7.0. The gluconic acid production by the resting cell system was more effective than those of the fermentation with respect to cultivation time, yield, recovery and re-use of the cell.l.l.l.l.l.l.

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Anti-proliferative Effect of Coptis Chinensis Extract in Hep G2 Cells

  • Kim, Jun-Lae;Oh, Se-Mi;Shin, Jang-Woo;Son, Jin-Young;Cho, Jung-Hyo;Lee, Yeon-Weol;Son, Chang-Gue;Cho, Chong-Kwan;Yoo, Hwa-Seung
    • The Journal of Korean Medicine
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    • v.27 no.4
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    • pp.48-56
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    • 2006
  • Objectives : This study is aimed to elucidate anti-hepatoma activity of Coptis Chinensis Extract (CCE) and evaluate its effect on proliferation of human hepatoma Hep G2 cells. Methods : To identify CCE and control the quality, we performed fingerprinting by high-performance thin layer chromatography (HPTLC). To investigate effects of CCE on anti-hepatoma activity, we measured cytotoxicity against Hep G2 cells compared with treatment of paclitaxel and 5-fluorouracil (5-FU). To examine the mechanism of inhibitory effect of CCE on Hep G2 cell proliferation, cell cycle distribution was evaluated using fluorescent activated cell sorter (FACS) Result : CCE showed a significant effect that arrests Hep G2 cells at the G2/M phase of the cell cycle. CCE combined with paclitaxel inhibited synergistically cell growth of Hep G2 cells. Conclusion : CCE may present anticancer effects through inhibition of hepatocellular carcinoma (HCC) cell proliferation via G2/M arrest, and may be a useful anticancer agent for HCC.

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Isolation of a Novel Gellan-Depolymerizing Bacillus sp. Strain YJ-1

  • Jung, Yu-Jin;Park, Cheon-Seok;Lee, Hyeon-Gyu;Cha, Jae-Ho
    • Journal of Microbiology and Biotechnology
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    • v.16 no.12
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    • pp.1868-1873
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    • 2006
  • A novel microorganism that could degrade high molecular weight gellan was screened and isolated from soil. On gellan plate, the microorganism grew well and completely liquefied the plate. The gellan-degrading microorganism was isolated by pure culture on glucose and nutrient agar medium afterwards. The 16S rDNA sequence analysis and biochemical tests using an API 50CHB/20E kit revealed that the strain belonged to Bacillus sp. The isolate, named as Bacillus sp. YJ-1, showed optimum gellan-degrading activity in 0.5% gellan medium at pH 7.5 and 37$^{\circ}C$. The activity was measured and evaluated by the thiobarbituric acid and thin-layer chromatography method. Mass spectrometry revealed that the major gellan.. depolymerized product was an unsaturated tetrasaccharide consisting of $\Delta$4,5-glucuronic acid-(1$\rightarrow$4 )-$\beta$-D-glucose-(1$\rightarrow$4)- $\alpha$-L-rhamnose-(1$\rightarrow$3)-$\beta$-D-glucose, which is a dehydrated repeating unit of gellan, thus the enzyme was identified as gellan lyase. When the gellan was present in the medium, the gellan-degrading activity was much higher than that in glucose-grown cells. These results indicate that in the presence of gellan, Bacillus sp. YJ-1 is able to metabolize the gellan by inducing gellan-degrading enzymes that can degrade gellan into small molecular weight oligosaccharides, and then the gellan. depolymerized products are taken up by the cells and utilized by intracellular enzymes.