• Title/Summary/Keyword: Thin Layer Chromatography(TLC)

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A STUDY ON ARACHIDONIC ACID METABOLISM OF CHRONIC PERIAPICAL LESIONS (만성 치근단주위 병소조직의 Arachidonic acid 대사에 관한 연구)

  • Park, Keum-Soon;Son, Ho-Hyun
    • Restorative Dentistry and Endodontics
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    • v.17 no.1
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    • pp.83-94
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    • 1992
  • This study was executed to measure the biosynthesis of arachidonic acid metabolic products in chronic periapical lesions, to compare the products among periapical granuloma, periapical cyst and chronic periapical abscess, and to understand the pathogensis of chronic periapical lesions. Tissues from 33 chronic periapical lesions of human teeth were enucleated during endodontic surgery. large part of each tissue was contained in liquid nitrogen immediately and the other was examined histologically. In histologically diagnosed 8 cases of periapical granuloma, 9 cases of periapical cyst and 8 cases of chronic periapical abscess. the tissues were homogenatecl and incubated with $_{14}C$-arachidonic acid. Lipid solvent extracts were separated by thin layer chromatography to be analyzed by autoradiography and TLC analyzer. 1. $TXB_2$, 6-keto-$PGF_1{\alpha}$ and $PGE_2$, $LTB_4$, HETEs, and unidentified product which are metabolic products of arachidonic acid were measured in the tissues of chronic peripaical lesions. 2. In all of periapical granuloma, cyst and abscess, the conversion rate of HETEs among all products was the highest(P<0.05), and the percentage of HETEs in total converted products was also the highest(P<0.05). 3. The concentration of each arachidonic acid product was higher in chronic periapical absecss than in periapical granuloma and cyst(P<0.05). The concentration of $TXB_2$ and HETEs in periapical cyst were hight than in periapical granuloma. 4. The relative amounts of total products from lipoxygenase pathway to those from cyclo-oxygenase pathway were about 7 fold in chronic periapical lesions. There was no difference among periapical granuloma, cyst and abscess(P<0.05). The total amount of products from each pathway were higher in chronic periapical abscess than in periapical cyst and granuloma.

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Quality Monitoring of Specification Standard of Gardeniae Fructus in the Korean Pharmacopoeia and Studies HPLC Standard Chromatogram (치자(梔子)의 규격 기준 모니터링 및 HPLC 표준크로마토그램 연구)

  • Kim, Kyoung Hee;Kim, Sun Mi;Shin, Seung Hoon;Lee, Young Jong;Baek, Wan Sook
    • The Korea Journal of Herbology
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    • v.32 no.2
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    • pp.97-105
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    • 2017
  • Objectives : Gardeniae Fructus is a ripe fruit of Gardenia jasminoides Ellis, which has been used as traditional medicines for anti-inflammatory, diuretic, antipyretic, and antibacterial activity. The aim of this study was to compare of Gardeniae Fructus in South Korea collected during three years according to the standards in monographs of the Korean Pharmacopoeia Eleventh edition (KP11). Methods : 30 items of Gardeniae Fructus from two cultivation regions were classified into dried(n=15) & steamed (n=15) and tested according to the standards in monographs of the KP11. Gardeniae Fructus was carried out identification(comparison of colors, thin layer chromatography), heavy metals, residual pesticides, total ash, and assay registered at KP11. Add to we tested loss on dry, contents of ethanol-soluble extracts, and HPLC profiling. Results : In TLC chromatogram of identification test, the spot of gardenoside and geniposide were observed at $R_f$ value of about 0.3 and 0.5. Heavy metals and residual pesticides met the requirements of the standards for all samples. The results of total ash of each samples are measured maximum 4.87 %. According to HPLC for assay, the samples contain 4.80~6.10 % of geniposide and 0.45~1.83 % of gardenoside. Conclusion : We have verified the current specification standard of Gardeniae Fructus and standard that is not set. By the results, it is proposed a new draft of loss on drying and confirmed the content of gardenoside revised. HPLC standard chromatogram of Gardeniae Fructus is proposed. We hope that it will help the standardization of Gardeniae Fructus.

Studies on Lipids in Fresh-Water Fishes 1. Distribution of Lipid Components in Various Tissues of Crucian Carp, Carassius carassius (담수어의 지질에 관한 연구 1. 붕어(Carassius carassius)의 부위별 지질성분의 분포)

  • CHOI Jin-Ho;RO Jae-Il;PYEUN Jae-Hyeong;CHOI Kang-Ju
    • Korean Journal of Fisheries and Aquatic Sciences
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    • v.17 no.4
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    • pp.333-343
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    • 1984
  • This study was designed to elucidate the lipid and its fatty acid composition in various tissues of fresh water fishes. The free and bound lipids in meat, skin and viscera of crucian carp (Carassius carassius) were extracted with ethyl ether and the mixed solvent of chloroform-methanol-water (10/9/1, v/v). The free and bound lipids were fractionated into neutral lipid, glycolipid and phospholipid by a silicic acid column chromatography using chloroform, acetone and methanol, respectively, and quantitatively analyzed by thin layer chromatography (TLC) and TLC scanner. The fatty acid compositions of polar ana nonpolar lipids in meat, and these of neutral lipid in various tissues were analyzed by gas liquid chromatography(GLC). The free lipid content in meat, skin and viscera was $6.22\%,\;9.95\%\;and\;9.76\%$, whereas the bound lipid content in those tissues was $10.01\%,\;3.56\%\;and\;7.36\%$, respectively. The neutral lipid contents in free lipid were ranged from $71.7\%$ to $89.4\%$, and $3{\sim}9$ times higher than those in bound lipid, while the phospholipid contents in bound lipid were ranged from $42.3\%$ to $63.2\%$, and $5{\sim}10$ times higher than those in free lipid. The neutral lipid was mainly consisted of triglyceride ($81.91{\sim}88.34\%$) in free lipid, and esterified sterol & hydrocarbon ($41.00{\sim}59.43\%$) in bound lipid. The phospholipid was mainly consisted of phosphatidyl ethanolamine($54.56{\sim}66.79\%$) and phosphatidyl choline ($21.88{\sim}34.28\%$) in free lipid, and phosphatidyl choline ($50.49{\sim}70.57\%$) and phosphatidyl ethanolamine ($15.74{\sim}24.92\%$) in bound lipid. The major fatty acids of polar lipid in free and bound lipids were $C_{16:0}\;(17.53\%,\;19.29\%)$, $C_{18:1}\;(24.57\%,\;16.08\%)$, $C_{18:2}\;(8.39\%,\;4.03\%)$, $C_{22:5}\;(1.68\%,\;8.08\%)$, and $C_{22:6}\;(6.22\%,\;13.60\%)$ and these of neutral lipid in free and bound lipids were $C_{16:0}\;(17.67\%,\;24.15\%)$, $C_{16:1}\;(12.81\%,\;5.52\%)$, $C_{18:1}\;(24.13\%,\;13.02\%)$, $C_{18:2}\;(15.47\%,\;8.68\%)$, $C_{22:5}\;(0.88\%,\;4.14\%)$ and $C_{22:6}\;(1.17\%,\;5.04\%)$, respectively. The unsaturations (TUFA/TSFA) of polar lipid in free and bound lipids were 2.02 and 2.74, and $1.5{\sim}2.0$ times higher than 1.51 and 1.23 of nonpolar lipid. In both polar and nonpolar lipids, w3 highly unsaturated fatty acid (w3HUFA) content of bound lipid was $2{\sim}5$ times higher than that of free lipid. The polyenoic acid contents such as $C_{20:5},\;C_{22:5}\;and\;C_{22:6}$ in bound lipid were $2{\sim}5$ times higher than these in free lipid. Consequently, there were significant difference between the lipid and its fatty acid composition in free and bound lipids and/or in various tissues.

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The study on the metabolism of benzidine in the isolated perfused rat liver (흰쥐의 적출 간 관류법을 이용한 벤지딘 대사에 관한 연구)

  • Bae, Mun Joo;Roh, Jae Hoon;Cho, Young Bong;Kim, Choon Sung;Chun, Mi Ryoung;Kim, Chi Nyon
    • Journal of Korean Society of Occupational and Environmental Hygiene
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    • v.6 no.1
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    • pp.28-37
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    • 1996
  • Benzidine, an aromatic amine used primarily in the manufacture of azo dyes, is recognized as a urinary bladder carcinogen in humans. In rats, mice, and hamsters, chronic exposure to benzidine resulted in tumors of the liver. The present study was undertaken to suggest analyzing the metabolites of benzidine with the optimal condition, identify the metabolites of benzidine, and observe time variance of the metabolites in the isolated perfusated rat liver. N-acetylbenzidine was synthesized by acetylation of benzidine with acetic anhydride and separated by thin layer chromatography(TLC) and high performance liquid chromatography(HPLC). To analysis benzidine and the metabolites of benzidine, HPLC operating condition has been optimized by means of preliminary experiment. The mobile phase consisted of acetonitrile(37%) in phosphate buffer, flow rate maintained at 1.0 ml/min. Optimal detective conditions were electrochemicaldetector(ECD) at 0.75 V for benzidine and N-acetylbenzidine and ultravioletdetector(UVD) at 287 nm for N,N'-diacetylbenzidine. The separation system was composed of a guard column and a separation column(Polymer C18, $4.6{\times}250cm$) at a temparature of $40^{\circ}C$. The perfusion system was equilibrated for 30 minutes before addition of benzidine to the perfusate. Samples of the perfusate were collected at time intervals(0, 10, 20, 30, 60, 90, 120 min) during the 2 hour perfusion. Before analyzing samples by HPLC/ECD/UVD, samples had been treated with sep-pak. Samples of perfusate analyzed by HPLC/ECD/UVD and the metabolites of benzidine in the isolated perfused rat liver were N-acetylbenzidine and N,N'-diacetylbenzidine. Benzidine metabolized over 60% during the initial 30 minutes of perfusion, extensively by 1 hour, and was undetectable in the perfusate. N-acetylbenzidine increased by 30 minutes of perfusion, declined. N,N'-diacetylbenzidine increased the 0-90 minutes period, remained constant during the 90-120 minutes period.

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A Comparative Study on the Lipid Components of Barley and Malt -I. Composition of Neutral Lipids- (보리와 맥아(麥芽)의 지방질(脂肪質) 성분(成分)에 관한 비교(比較) 연구 -제(第)1보(報) : 중성지질(中性脂質)의 조성(組成)-)

  • Shin, Hyo-Sun;Lee, Kang-Hyon;Lee, Sang-Young
    • Korean Journal of Food Science and Technology
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    • v.13 no.1
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    • pp.30-36
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    • 1981
  • Neutral lipids of the total lipid extracted from 4 representative varieties of barley grown in Korea and their corresponding malt were studied. Total lipids of barley and malt were solvent extracted with chloroform:methanol:water (1.0 : 1.0 : 0.9, v/v), The total lipids were fractionated into neutral and polar lipids by silicic acid column chromatography, and neutral lipids fraction was separated by thin layer chromatography and quantitated by TLC scanner. The fatty acid compositions was determined by gas liquid chromatography. The average content of total lipid in the 4 barleys and their malts were 3.3 and 2.9%, and average of neutral lipids content in the barley and malt lipids were 73.8 and 68.5%, respectively. Among the neutral lipids, triglycerides and free fatty acids were the major components, and triglycerides content decreased and free fatty acids content increased during malting. Sterol esters, free sterols, 1,3-and 1,2-diglycerides were the minor components of the neutral lipids, and contents of those components showed increasing tendency during malting. The major fatty acid composition of the total lipids were linoleic, palmitic and oleic acids, and in general, the malts had lower amounts of unsautrated fatty acids and high amounts of saturated fatty acids. Fatty acid composition of neutral lipids was of almost the same pattern as that of the total lipids.

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Collection of Soil Actinomycetes from Cheju Island and Screening for their Antibacterial Activities (제주도 토양방선균의 수집과 항균물질 생산균의 선별)

  • Chung, Wan-Seok;Kim, Chang-Jin;Ko, Young-Hwan
    • Applied Biological Chemistry
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    • v.42 no.2
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    • pp.99-104
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    • 1999
  • Soil actinomycetes of 703 strains were isolated from 25 sampling points in Cheju Island using 4 different media. Arginine glycerol salts agar containing soil extract was found to be the best medium for the isolation of soil actinomycetes. Soil samples from pasture land showed higher population and diversity of the actinomycetes than those from citrus field, forest, island, hill or valley. When the antibacterial activity of the 526 isolates was tested against three bacterial strains, Escherichia coli, Staphylococcus aureus and Pseudomonas solanacearum the frequency of the isolates with antibacterial activity varied much depending upon the media used for isolation and cultivation. BL106Ba, one of the 10 isolates that showed antibacterial activity against all the above 3 test strains, was chosen based upon the pH and heat stability of its antibacterial metabolites, and was identified as Streptomyces sp. based upon its cultural, morphological and physiological characteristics. The partially purified white crystalline substance obtained from the culture supematant of BL1063a through cation exchange chromatography(AG MP-50) and three times consecutive gel filtration(Sephadex G-10) showed high antimicrobial activity against gram positive and negative bacteria, but low activity against yeasts. The partially purified substance was found to contain at least four different compounds with antibacterial activity by both thin layer chromatography and high performance liquid chromatography.

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Triglyceride Composition of Pine Nut Oil (잣기름의 Triglyceride조성(組成))

  • Chun, Suck-Jo;Park, Yeung-Ho
    • Korean Journal of Food Science and Technology
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    • v.16 no.2
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    • pp.179-181
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    • 1984
  • The present study was directed to define the triglyceride composition of pine nut oil. The triglycerides were separated from pine nut oil by thin layer chromatography, and fractionated by high performance liquid chromatography on the basis of partition numbers. Each of these collected fractions were fractionated again by gas liquid chromatography (GLC) according to the acyl carbon number of the triglyceride, and fatty acid composition of the triglyceride was also analyzed by GLC. The pine nut oil consisted of thirty two kinds of triglycerides, and the major triglycerides of pine nut oil were those of $(C_{18:2},\;C_{18:2},\;C_{18:3}\;;\;34.9%)$, $(C_{18:1},\;C_{18:2},\;C_{18:3}\;;\;10.8%)$, $(C_{18:1},\;C_{18:1},\;C_{18:2}\;;\;9.9%)$, $(C_{18:1},\;C_{18:1},\;C_{18:1}\;;\;6.5%)$, $(C_{18:1},\;C_{18:1},\;C_{18:2}\;;\;6.3%)$, $(C_{18:1},\;C_{18:1},\;C_{18:3}\;;\;4.8%)$, $(C_{16:0},\;C_{18:2},\;C_{18:3}\;;\;3.3%)$, $(C_{18:0},\;C_{18:1},\;C_{18:2}\;;\;2.7%)$, $(C_{16:0},\;C_{18:1},\;C_{18:2}\;;\;2.6%)$, $(C_{16:0},\;C_{18:2},\;C_{18:2}\;;\;2.2%)$, $(C_{16:0},\;C_{18:1},\;C_{18:3}\;;\;1.9%)$, $(C_{16:0},\;C_{18:2},\;C_{18:2}\;;\;1.7%)$, $(C_{16:0},\;C_{18:1},\;C_{18:1}\;;\;1.7%)$, $(C_{18:1},\;C_{18:3},\;C_{18:3}\;;\;1.5%)$.

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Screening of Natural Products for Anti-diabetic Activity and Analysis of Their Active Compounds (항당뇨 효능이 있는 천연물의 탐색 및 활성물질의 분석)

  • Hwa Sin Lee;Bo Bae Park;Sun Nyoung Yu;Min Ji Kim;Yun Jin Bae;Yi Rooney Lee;Ye Eun Lee;Si Yoon Kim;Yun Ho Shim;Soon Cheol Ahn
    • Journal of Life Science
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    • v.33 no.10
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    • pp.783-790
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    • 2023
  • Modern people have an increased incidence of metabolic diseases due to changed eating habits, and diabetes is considered the most significant metabolic disease. Given that existing diabetes treatments are accompanied by side effects, the aim of this study was to identify traditional natural products that have anti-diabetic activity. The potential anti-diabetic and antioxidant activities of natural products were examined using 2,2-diphenyl-1-picrylhydrazyl (DPPH) free radical scavenging assay, α-glucosidase assay, and protein tyrosine phosphatase 1B (PTP1B) inhibition assay. Methanol extracts of Ulmus davidiana var. japonica, Acer tegmentosum branches, Nelumbo nucifera seeds, and Carthamus tinctorius seeds were found to have high anti-diabetic activity and further fractionated with solvents using ethyl acetate and butanol. Consequently, the ethyl acetate fraction of C. tinctorius seeds (MG-11-E) with high α-glucosidase and PTP1B inhibitory activity was selected. MG-11-E was subjected to preparative thin layer chromatography, and fraction #6 showed high α-glucosidase and PTP1B inhibitory activity. Fraction #6 was analyzed and fractionated via high performance liquid chromatography with 50% methanol as the mobile phase, and anti-diabetic activity was observed in the sample that eluted after 4 min as a single peak. The α-glucosidase inhibitory activity exhibited by this sample seemed to be greater than the PTP1B inhibitory activity; thus, it was concluded that a greater anti-diabetic therapeutic effect may be achieved by combining this agent with natural products that inhibit PTP1B activity.

Characterization of β-agarase from Isolated Simiduia sp. SH-4 (분리된 Simiduia sp. SH-4가 생산하는 β-agarase의 특성조사)

  • Kim, Jae-Deog;Lee, Sol-Ji;Jo, Jeong-Gwon;Lee, Dong-Geun;Lee, Sang-Hyeon
    • Journal of Life Science
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    • v.26 no.4
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    • pp.453-459
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    • 2016
  • Agarases are classified into α-agarase and β-agarase that produce agarooligosaccharides and neoagarooligosaccharides, respectively. Neoagarooligosaccharides have whitening effect of skin, delay of starch degradation, and inhibition of bacterial growth etc. Hence, the object of this study was to isolate a novel agarase producing marine bacterium and characterization of its β-agarase. A novel agar-degrading bacterium was isolated from seashore of Namhae at Gyeongnamprovine, Korea and purely cultured with Marine agar 2216 media. The isolated bacterium was identified as Simiduia sp. SH-4 after 16S rRNA gene sequencing. The enzymatic sample was obtained from culture media of Simiduia sp. SH-4. Enzymatic activity was highly increased from 20(30% relative activity) to 30℃ (100%) and decreased from 30 to 40℃(75%) and so more. Relative activity was 100% at pH 6 while those were about 91% and 59% at pH 5.0 and 7.0, respectively, meaning the enzyme possesses narrow optimal pH range. Hence, the enzyme exhibited the maximal activity with 120.4 units/l at pH 6.0 and 30℃ in 20 mM Tris-HCl buffer. Thin layer chromatography (TLC) analysis showed that Simiduia sp. SH-4 produces β-agarase, which hydrolyze agarose to produce biofunctional neoagarooligosaccharides such as neoagarotetraose and neoagarobiose. Hence, broad applications would be possible using Simiduia sp. SH-4 and its enzyme in the food industry, cosmetics and medical fields.

Antioxidative and Cytotoxicity Activities against Human Colon Cancer Cells Exhibited by Edible Crude Saponins from Soybean Cake (대두박 식용사포닌의 항산화 및 대장암세포 성장 억제효과)

  • Park, Kyung-Uk;Kim, Jae-Yong;Seo, Kwon-Il
    • Food Science and Preservation
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    • v.16 no.5
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    • pp.754-758
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    • 2009
  • To develop soybean cake as a functional food material, the anti-oxidative and cytotoxic activities against human colon cancer cells of crude saponins isolated from 70% (v/v) ethanol extracts of cake were investigated. The Diaion HP-20 adsorption method was used for isolation of crude saponins, which were then eluted with 100% ethanol. The non-saponin fraction was removed by elution with $H_2O$ and 20% (v/v) ethanol. The results of thin layer chromatography (TLC) analysis confirmed that crude saponins were present in the 100% ethanol extract of soybean cake. The hydrogen-donating properties of saponins were more than 60% at a concentration of $1,000\;{\mu}g/mL$. malondialdehyde(MDA) production was $1,200\;{\mu}mol\;MDA/g$ in mouse liver homogenate treated with crude saponins at the concentration of $1,000\;{\mu}g/mL$. This value was lower than that of the control, which was $3,700\;{\mu}mol\;MDA/g$. Saponins inhibited the growth of colon cancer cells in a dose- and time-dependent manner. Saponins also resulted in a decrease in the proportion of cells in the G1 phase of the cell cycle, whereas the cell proportion in G2/M phase was increased with $1,000\;{\mu}g/mL$ saponins. Thus, we conclude that saponins may induce G2/M cell cycle arrest.