• Title/Summary/Keyword: thin layer chromatography (TLC)

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Hygienic Studies on Inactive Substances of Sterigmatocystin by Artificial Gastric Juice (인공(人工) 위액(胃液)에 의한 Sterigmatocystin의 실활물(失活物)에 관한 위생화학적(衛生化學的) 연구(硏究))

  • Kim, Soo-Yeon;Oh, You-Jin;Yeo, Sin-Koo;Jang, Seong-Jae
    • The Korean Journal of Mycology
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    • v.14 no.1
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    • pp.61-70
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    • 1986
  • Sterigmatocystin bears a close structural relationship to aflatoxin $B_1$ and is a carcinogenic compound that has been shown to affect various species of experimental animals. Reaction and toxicity of sterigmatocystin in the artificial gastric juice were investigated. Sterigmatocystin was degraded in artificial gastric juice and extracted by the method of A.O.A.C. After cleaned up by silica gel column chromatography, this substance was detected and characterized by thin layer chromatography, UV, IR and mass spectra. It showed $R{\mathcal{f}}$ 0.4 and brick-red color by TLC. Especially, in the mass spectrum of it, fragment peak at m/e 327 was due to the loss of the $-CH_3$ and $-H_2O$, fragment peak at m/e 341 was due to the loss of the $H_2O$ and $-H^+$, and fragment peak at m/e 239 was due to the loss of the 2-chloro-tetrahydrofuran and methyl group from the parent molecule. Therefore, a degraded substance of sterigmatocystin reacted in artificial gastric juice (Sub. K) was estimated with additional formation of hydrochloric acid. In four-day-old chicken embryos, the mean lethal dose $(LD_{50})$ was $140\;{\mu}g/egg$, and 90 to 100% of the embryos were killed with 1 mg/egg. This $LD_{50}$ $140\;{\mu}g/egg$ compared with an $LD_{50}$ $14.69\;{\mu}g/egg$ for sterigmatocystin (acute toxicity) showed the substance to be much less toxic than sterigmatocystin.

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Production of Minor Gisenosides from Gypenoside V (Gypenoside V로부터 minor ginsenosides의 생산)

  • Son, Na-Ri;Min, Jin-Woo;Jang, Mi;Kim, Hyo-Yeon;Jeon, Ji-Na;Yang, Deok-Chun
    • Proceedings of the Plant Resources Society of Korea Conference
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    • 2010.10a
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    • pp.20-20
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    • 2010
  • Panax ginseng C.A Meyer is frequently taken orally as a traditional herbal medicine in Asian countries. The major components of ginseng are ginsenoside, which are pharmaceutical activity. The six major ginsenosides, including Rb1, Rb2, Rc, Rd, Re and Rg1 account for 90% of total ginsenosides. Even though the minor ginsenosides, including Rg3, Rh2 and compound K has high pharmacetical activities, the price of minor ginsenosides is too high. Therefore we isolated the gypenoside V and made it converted to minor ginsenosides. In the plant Gynostemma pentaphyllum Makino, gypenosdie V was presented as dominant saponin (content about 2.4%), and was similar to protopanaxadol type ginsenosides such as ginsenoside Rb1. In this study, we confirmed that the coversion of gypenoside V to minor ginsenosides after using the various treatment such as heating, acid treatment, commercial edible enzyme, and lactobacillus. Consequently, we optimizied the transformation of gypenoside V to minor ginsenoside using Thin Layer Chromatography (TLC), High Performance Liquid Chromatography (HPLC), Time-of-flight Mass Spectrometry (LC/TOF/MS).

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Purification of $\beta$-glucanase from Bacillus subtilis Using Chromogenic Substrate (색소기질을 이용한 Bacillus subtilis의 $\beta$-glucanase 정제)

  • 이성택;양진오;정안식
    • Korean Journal of Microbiology
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    • v.26 no.3
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    • pp.223-229
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    • 1988
  • Bacillus subtilis K-4-3, which produces considerable amount of $\beta$-glucanase was selected among extracellular $\beta$-glucanase-producing bacteria isolated from soil. $\beta$-glucanase was purified by ammonium sulfate fractionation, Sephadex G-100 gel filtration and DEAE-sephacel ion exchange chromatography. The purified enzyme revealed a single band by polyacrylamide gel electrophoresis and SDS-polyacrylamide gel electrophoresis. Its molecular weight was estimated to be 17000 dalton by SDS-polyacrylamide gel electrophoresis. The optimum pH and temperature of the purified $\beta$-glucanase were 7.0 and $50^{\circ}C$, respectively. The enzyme was strongly inhibited by 1.0mM of $Fe^{3+}$, and activated by 1.0mm of $Li^{}47+$. The absence of glucose after thin layer chromatography of reaction products revealed that the purified enzyme contains no cellobiase or laminarinbiase activity. The loberation of ki, tri-and tetra-saccharide as reaction products can be explained by endoaction of the enzyme.

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Evaluation of Antioxidant Potential and UV Protective Properties of Four Bacterial Pigments

  • Rupali Koshti;Ashish Jagtap;Domnic Noronha;Shivali Patkar;Jennifer Nazareth;Ruby Paulose;Avik Chakraborty;Pampi Chakraborty
    • Microbiology and Biotechnology Letters
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    • v.50 no.3
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    • pp.375-386
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    • 2022
  • In the present study, four distinctly colored bacterial isolates that show intense pigmentation upon brief ultraviolet (UV) light exposure are chosen. The strains are identified as Micrococcus luteus (Milky yellow), Cryseobacterium pallidum (Yellow), Cryseobacterium spp. (Golden yellow), and Kocuria turfanensis (Pink) based on their morphological and 16S rDNA analysis. Moderate salinity (1.25%), 25-37℃ temperature, and pH of 7.2 are found to be the most favorable conditions of growth and pigment production for all the selected isolates. The pigments are extracted using methanol: chloroform (1:1) and the purity of the pigments are confirmed by high-performance liquid chromatography (HPLC) and thin-layer chromatography (TLC). Further, Fourier transform infrared (FTIR) and UV-Visible spectroscopy indicate their resemblance with carotenoids and flexirubin family. The antioxidant activities of the pigments are estimated, and, all the pigments have shown significant antioxidant efficacy in 2,2'-azino-bis(3-ethylbenzothiazoline-6-sulfonic acid) (ABTS), 2,2-diphenyl-1-picryl-hydrazyl (DPPH), and ferric reducing antioxidant power (FRAP) assays. The UV protective property of the pigments is determined by cling-film assay, wherein, at least 25% of UV sensitive Escherichia coli survive with bio-pigments even after 90 seconds of UV exposure compared to control. The pigments also hold a good sun protective factor (SPF) value (1.5-4.9) which is calculated with the Mansur equation. Based on these results, it can be predicted that these bacterial pigments can be further developed into a promising antioxidant and UV-protectant for several biomedical applications.

Saponins of Korean Ginseng Panax ginseng C.A. Meyer (Part III) -Saponins of ginseng by the cultivating locations, sampling seasons, plant parts, growing stages and the processings- (한국인삼(韓國人蔘)의 Saponin에 관(關)한 연구(硏究)제3보(第三報) -산지별(産地別), 부위별(部位別), 재배기간별(栽培期間別) 인삼(人蔘) 및 가공중(加工中) Saponin함량(含量)에 관(關)하여-)

  • Cho, Sung-Hwan
    • Applied Biological Chemistry
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    • v.20 no.2
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    • pp.188-204
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    • 1977
  • The studies on the saponins of Korean ginseng, Panax ginseng C.A. Meyer, were performed according to the cultivating locations, sampling seasons, plant parts, and growing stages. The changes in saponin content in the course of manufacturing Red ginseng and Ginseng extract were observed. In this paper, a new method for the determination of the total and the individual saponin glucosides was proposed and applied to the samples under study. The method employing Digital Densitorol DMU-33C (Toyo electric Co., Japan) followed the separation of the saponins by means of a preparative thin layer chromatography. The saponin contents and their fractional distribution were summarized as follows: 1. The average concentrations(% plant dry weight) of semi-purified saponins in the roots of Korean ginseng planted in the various locations were 5.0%(Keumsan), 6.0% (Kimpo), and 5.4% (Pocheon), respectively. 2. There were 3.3% saponins in White ginseng(Rhizome) and 12.7% saponins in Ginseng tail (Fibrous root). 3. Regarding the year of growth, the contents of saponins were 90.3mg (2-year-old ginseng), 254.4mg (3-year-old ginseng), 404.2mg (4-year-old ginseng). 999.6mg (5-year-old ginseng), and 1377.1mg (6-year-old ginseng) respectively, and the saponin factions containing panaxatriol as an aglycone increased. 4. Thin layer chromatography revealed that Red ginseng yielded many saponins which Shibata et al. designated as $ginsenoside-Rb_1$ (22.1%), $-Rb_2(15.4%)$, -Rc(12.6%), -Re (15.7%), and $-Rg_1$, (9.3%). 5. 29.9% of crude saponins were isolated from ethanolic extract of Panax ginseng fibrous root and their extraction yield was 94.2% of fibrous root saponin.

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A Simple Method for Evaluation of Pepper Powder Color Using Vis/NIR Hyperspectral System (Vis/NIR 초분광 분석을 이용한 고춧가루 색도 간이 측정법 개발)

  • Han, Koeun;Lee, Hoonsoo;Kang, Jin-Ho;Choi, Eunah;Oh, Se-Jeong;Lee, Yong-Jik;Cho, Byoung-Kwan;Kang, Byoung-Cheorl
    • Horticultural Science & Technology
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    • v.33 no.3
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    • pp.403-408
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    • 2015
  • Color is one of the quality determining factors for pepper powder. To measure the color of pepper powder, several methods including high-performance liquid chromatography (HPLC), thin layer chromatography (TLC), and ASTA-20 have been used. Among the methods, the ASTA-20 method is most widely used for color measurement of a large number of samples because of its simplicity and accuracy. However it requires time consuming preprocessing steps and generates chemical waste containing acetone. As an alternative, we developed a fast and simple method based on a visible/near infrared (Vis/NIR) hyperspectral method to measure the color of pepper powder. To evaluate correlation between the ASTA-20 and the visible/near infrared (Vis/NIR) hyperspectral methods, we first measured the color of a total of 488 pepper powder samples using the two methods. Then, a partial least squares (PLS) model was postulated using the color values of randomly selected 3 66 samples to predict ASTA values of unknown samples. When the ASTA values predicted by the PLS model were compared with those of the ASTA-20 method for 122 samples not used for model development, there was very high correlation between two methods ($R^2=0.88$) demonstrating reliability of Vis/NIR hyperspectral method. We believe that this simple and fast method is suitable for highthroughput screening of a large number of samples because this method does not require preprocessing steps required for the ASTA-20 method, and takes less than 30 min to measure the color of pepper powder.

Identification and Biochemical Characterization of a New Xylan-degrading Streptomyces atrovirens Subspecies WJ-2 Isolated from Soil of Jeju Island in Korea (제주도 토양으로부터 자일란 분해 Streptomyces atrovirens subspecies WJ-2 동정 및 효소의 생화학적 특성 규명)

  • Kim, Da Som;Bae, Chang Hwan;Yeo, Joo Hong;Chi, Won-Jae
    • Microbiology and Biotechnology Letters
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    • v.44 no.4
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    • pp.512-521
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    • 2016
  • A bacterial strain was isolated from a soil sample collected on Jeju Island, Korea. The strain, designated WJ-2, exhibited a high xylanase activity, whereas cellulase activity was not detected. The 16S rRNA gene sequence of WJ-2 was highly similar to type strains of the genus Streptomyces. A neighbor-joining phylogenetic tree based on 16S rRNA gene sequences showed that strain WJ-2 is phylogenetically related to Streptomyces atrovirens. Furthermore, DNA-DNA hybridization analysis confirmed that strain WJ-2 is a novel subspecies of Streptomyces atrovirens. The genomic DNA G+C content was 73.98 mol% and the major fatty acid present was anteiso-C15:0 (36.19%). The growth and xylanase production of strain WJ-2 were significantly enhanced by using soytone and xylan as nitrogen and carbon sources, respectively. Crude enzyme preparations from the culture broth of strain WJ-2 exhibited maximal total xylanase activities at pH 7.0 and $55^{\circ}C$. Thin-layer chromatography analysis revealed that the crude enzyme degrades beechwood xylan to yield xylobiose and xylotriose as the principal hydrolyzed end products.

Constructing Database for Drugs and its Application to Biological Sample by HPTLC and GC/MS (HPTLC와 GC/MS를 이용한 의약품의 데이타베이스화 및 생체시료에의 응용)

  • Yoo, Young-Chan;Park, Sung-Woo;Lim, Mie-Ae;Baeck, Seung-Kyung;Park, Seh-Youn;Lee, Ju-Seon;Lho, Dong-Seok
    • Analytical Science and Technology
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    • v.13 no.2
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    • pp.136-150
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    • 2000
  • For the identification of unknown drugs in biological samples, we attempted rapid high performance thin layer chromatographic method which is sensitive and selective chromatographic analysis of high performance thin layer chromatography (HPTLC) with automated TLC sampler and ultra-violet (UV) scanner. We constructed HPTLC database (DB) on two hundred five drugs by using the data of Rf values and UV spectra (scan 200-360 nm) as well as gas chromatography/mass spectrometry (GC/MS) DB on ninety six drugs by using the data of relative retention time (RRT) on lidocain and mass spectra. After extracting drugs in biological sample by solid phase extraction (Clean Screen ZSDAU020), we applied them to HPTLC and GC/MS DB. Drugs, especially extracted from biological samples, showed good matching ratio to HPTLC DB and these drugs were confirmed by GC/MS. In conclusion, this DB system is thought to be very useful method for the screening of unknown drugs in biological samples.

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Isolation and Characterization of Lactobacillus brevis AML15 Producing γ-Aminobutyric acid ((γ-Aminobutyric acid를 생산하는 Lactobacillus brevis AML15의 분리 및 특성)

  • Shin, Ji-Won;Kim, Dong-Geol;Lee, Yong-Woo;Lee, Hyoung-Seok;Shin, Kee-Sun;Choi, Chung-Sig;Kwon, Gi-Seok
    • Journal of Life Science
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    • v.17 no.7 s.87
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    • pp.970-975
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    • 2007
  • For the screening of ${\gamma}-aminobutyric$ acid (CABA)-producing bacteria, 86 bacterial strains which produce GABA were isolated from Kimchi and Salted fisk .Among these, three strains designated AML15, AML45-1, AML72 with relatively high GABA productivity were selecled by thin layer chromatography (TLC). To elucidate the relationship between isolated strains and the genus Lactobacillus, their 16S rDNA sequence were examined. The result of their DNA sequences showed 99% similarity with Lactobacillus brevis ATCC 367. On the basis of the these results, isolated strains were identified as Lactobacillus brevis and designated L. brevis AML15. In order to determine the optimum conditions for GABA production, the isolated strains were cultivated in pyridoxal phosphate (PLP) and monosodium glutami. acid (MSG). Results showed that L. brevis AML15 had the highest CABA productivity with 10,424 $nM/{\mu}l$ concentration in MRS broth containing 5% (w/v) MSG and 10 ${\mu}M$ PLP at pH 5.0. The results imply that L. brevis AML15 has the potential to be developed as a strain for GABA hyper-production.

Isolation of Bacillus subtilis GS-2 Producing γ-PGA from Ghungkukjang Bean Paste and Identification of γ-PGA (청국장으로부터 분리한 Poly(γ-glutamic acid)를 생산하는 균주 Bacillus subtilis GS-2의 분리 및 γ-PGA의 확인)

  • Bang, Byung-Ho;Jeong, Eun-Ja;Rhee, Moon-Soo;Kim, Yong-Min;Yi, Dong-Heui
    • Journal of Applied Biological Chemistry
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    • v.54 no.1
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    • pp.1-6
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    • 2011
  • ${\gamma}$-PGA(poly-${\gamma}$-glutamic acid) is an unusual anionic polypeptide that is made of D- and L-glutamic acid units connected by amide linkages between ${\alpha}$-amino and ${\gamma}$-carboxylic acid groups. ${\gamma}$-PGA has been isolated from many kinds of organisms. Many Bacillus strains produce ${\gamma}$-PGA as a capsular material of an extracellular viscous material. It is safe for eating as a viscosity element of fermented soybean products such as Chungkookjang and Natto. It is biodegradable, edible and nontoxic toward humans and the environment and its molecular weight varies from ten thousand to several hundred thousand depending on the kinds of strains used. Therefore, potential applications of ${\gamma}$-PGA and its derivatives have been of interest in the past few years in a broad range of industrial fields such as food, cosmetics, medicine, water-treatment, etc. In this study, a bacterium, Bacillus subtilis GS-2 isolated from the Korean traditional seasoning food, Chungkookjang could produce a large amount of ${\gamma}$-PGA with high productivity and had a simple nutrient requirement. Based on carbon utilization pattern and partial 16S rRNA sequence analysis, the GS-2 strain was identified as B. subtilis. The determination of purified ${\gamma}$-PGA was confirmed with thin layer chromatography (TLC), high performance liquid chromatography (HPLC), fourier transform infrared (FT-IR) spectra, and $^1H$-nuclear magnetic resonance ($^1H$-NMR) spectroscopy.