• Title/Summary/Keyword: reversed phase

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Isolation and Quantitative Analysis of Betulinic Acid and Alphitolic Acid from Zyziphi Fructus (대추로부터 베튜리닉 산과 알피톨릭 산의 분리 및 정량)

  • Bae, Gi-Hwan;Lee, Sang-Myeong;Lee, Eun-Sil;Lee, Jun-Seong;Gang, Jong-Seong
    • YAKHAK HOEJI
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    • v.40 no.5
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    • pp.558-562
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    • 1996
  • Betulinic acid and alphitolic acid, the triterpenoids of Zyziphi Fructus, were isolated with silica gel column chromatography and used as the standard substances for the analysis. The compounds were determined with HPLC and HPTLC. They were separated on reversed phase column (Nova-Pak C18) with 0.05M Na2HPO4-methanol (19:81) in HPLC and detected at 210nm. Separation on HPTLC precoated silica gel F254 plates was carried out with chloroform-methanol (6:1) and the separated compounds were reacted with p-anisaldehyde and detected at 540nm. The contents of betulinic acid and alphitolic acid in Zyziphi Fructus from four different regions in Korea were in the range of 2.9~3.8% and 3.2~3.9%, respectively.

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Determination of Costunolide from Aucklandiae Radix and Substitutive Herbs by Reversed-Phase HPLC (HPLC를 이용한 목향 및 유사 한약재에 함유된 Costunolide 비교)

  • Oh, Ju-Hee;Hong, Seon-Pyo;Choi, Ho-Young;Park, Yong-Ki;Lee, Je-Hyun
    • The Korea Journal of Herbology
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    • v.23 no.3
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    • pp.61-66
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    • 2008
  • Objecives : Aucklandiae Radix is a root of Aucklandia lappa which has been widely used for regulating the flow of vital energy, invigorating the spleen, alleviating pain. Aucklandiae Radix contains the costunolide which is the main ingredient. The substitutive Aucklandiae Radix are Inulae helenii Radix, Aristolchiae Radix, Vladimiriae Radix, and Inulae racemosi Radix in Korea and China. This paper is analysised and compared the costunolide and HPLC pattern in Aucklandiae Radix and substitute herbs. Methods : Chromatographic separation performed using C18 column(Luna 5 u, 250 mm ${\times}$ 4.6 mm) with a mixture of methanol and water(65:35)(v/v). The analyses detected at UV(210 nm). Results : Optimal extraction condition of costunolide was 100% methanol for 2hr. Costunolide was detected in Aucklandiae Radix and Vladimiriae Radix, but other herbs were not detected. In Korea herbal market, Aristolchiae Radix merchandise was identified as the imported Inulae helenii Radix. Conclusions : According to above results, this method was useful identified to Aucklandiae Radix and substitutive herbs. In Korea herbal market, Aristolchiae Radix was identified as Inulae helenii Radix.

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The Separation and Determination of Rare Earth Elements by Ion-Association Chromatography (희토류 원소의 분리 및 정량을 위한 이온회합 크로마토그래피)

  • Lee, Seung Hwa;Lee, Cheol;Jeong, Koo Soon
    • Journal of the Korean Chemical Society
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    • v.34 no.1
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    • pp.69-75
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    • 1990
  • An ion-association chromatography was applied for the separation and determination of individual rare earth elements (REE) contained in mineral monazite. Prior to the determination, the group separation of REE was achieved by a cation exchange column of Dowex 5OW-X8 resin. The quantitative recovery of REE by the resin column, free from coexisting elements in monazite, was confirmed with radioactive tracers as well as with ICP-MS. Individual REE at ppm level was separated on reversed-phase column ($\mu$-Bondapak $C_{18}$) using gradient elution from 0.05 to 0.3 M $\alpha$-hydroxyisobutyric acid at pH 4.6. The individual REE was detected at 546 nm following post-column reaction with PAR (4-(2-pyridylazo)-resorcinol monosodium salt).

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Two Biophenolic Glycosides from Portulaca oleracea (쇠비름에서 분리된 2개의 Biophenolic Glycosides)

  • Youngwan Seo;Jongheon Shin;Burm Jong Lee;Dong Seok Lee
    • Journal of the Korean Chemical Society
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    • v.47 no.1
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    • pp.43-46
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    • 2003
  • From Portulaca oleracea which is widely distributed in Korea and has long been used as a folk medicine, two biophenolic glycosides, 3-hydroxy-1-(2-hydroxyethyl)phenyl-4-O-${\beta}$-D-glucopyranoside (1) and 2-(3,4-dihydroxyphenyl) ethyl-O-${\beta}$-D-glucopyranoside (2) were isolated using column chromatography and reversed-phase HPLC. $^{13}C$ NMR spectral assignment for these compounds was revised by the extensive 2-D NMR experiments such as NOESY, HMQC, and HMBC. These compounds showed a considerable antioxidant effect in DPPH assay system.

Effects of Climatic Factors and Tapping Date on Yield and Quality of Lactree (Rhus verniciflua) Sap (기상요인과 채취시기가 옻나무 칠액채취량 및 칠액의 質에 미치는 영향)

  • 김만조
    • Korean Journal of Plant Resources
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    • v.11 no.1
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    • pp.70-79
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    • 1998
  • This study was conducted to understand the effects of weather and tapping date on yield and quality of lactree(Rhus verniciflua) sap yield showed a significant positive correlation with the minimum temperature of one day before sap collection at 1% level and with theminimum humidity of theday of sap collection at 5% level. However, the differences between the maximum and the minimum temperatures and humidities of the day of sap collection were negatively correlated with the sap yield at 5% level. Multiple regression analysis indicated that the minimum temperature of one day bofore sap collection and the minumum humidity of the day sap collection were important factors for increasing sap yield. The high sap yield of lactree by Japanese tapping method was recorded during mid-July and early August. Seasonal variation in lactree sap constituents was observed. The sap collected on 15th of August contained the highest urushiol content (68.3%) and the lowest water content resulting in high quality of lactree sap. By reversed-phase HPLC analysis, fove urushiol components were separated from each other depending on the number of doulbe bonds in the side-chain , and seaxonal variation of urushiol composition was noticed. The 3-C15 triene content fo the sap collected on 5th of July was the highest(77.56%) indicating the major component of urushiol which affects lactree sap quality.

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Determination of Recombinant Human Epidermal Growth factor (rhEGF) in a Pharmaceutical Formulation by High Performance Liquid Chromatography with Electrochemical Detection

  • Lee, Kang-Woo;Hwang, Kyung-Hwa;Kim, Chang-Soo;Han, Kun;Chung, Youn-Bok;Park, Jeong-Sook;Lee, Yong-Moon;Moon, Dong-Cheul
    • Archives of Pharmacal Research
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    • v.24 no.4
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    • pp.355-359
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    • 2001
  • A novel HPLC method with electrochemical detection has been developed for the determination of recombinant human epidermal growth factor (rhEGF) in pharmaceutical products. rhEGF was separated from other components in formulation on a reversed-phase C18 column with 24% acetonitrile in 0.1 M phosphate buffer (pH 4.75). The optimum electrochemical oxidation of EGF was obtained at 0.85 V vs. Ag/AgCl in a glassy carbon working electrode due to electroactive tyrosine, tryptophan, methionine, and arginine residues. The quantitation range was from 1.0 to 200 ng of rhEGF with the linear correlation coefficient greater than 0.999. The method was successfully applied for the quantitation of rhEGF in a pharmaceutical preparation.

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Phosphate and Carbon Source Regulation of Alkaline Phosphatase and Phospholipase in Vibrio vulnificus

  • Oh, Wan-Seok;Im, Young-Sun;Yeon, Kyu-Yong;Yoon, Young-Jun;Kim, Jung-Wan
    • Journal of Microbiology
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    • v.45 no.4
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    • pp.311-317
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    • 2007
  • In this study, the effects of phosphate concentration and carbon source on the patterns of alkaline phosphatase (APase) and phospholipase (PLase) expression in Vibrio vulnificus ATCC 29307 were assessed under various conditions. The activities of these enzymes were repressed by excess phosphate (4 mM) in the culture medium, but this repression was reversed upon the onset of phosphate starvation in low phosphate defined medium (LPDM) containing 0.2 mM of phosphate at approximately the end of the exponential growth phase. The expressions of the two enzymes were also influenced by different carbon sources, including glucose, fructose, maltose, glycerol, and sodium acetate at different levels. The APase activity was derepressed most profoundly in LPDM containing fructose as a sole carbon source. However, the repression/derepression of the enzyme by phosphate was not observed in media containing glycerol or sodium acetate. In LPDM-glycerol or sodium acetate, the growth rate was quite low. The highest levels of PLase activity were detected in LPDM-sodium acetate, followed by LPDM-fructose. PLase was not fully repressed by high phosphate concentrations when sodium acetate was utilized as the sole carbon source. These results showed that multiple regulatory systems, including the phosphate regulon, may perform a function in the expression of both or either APase and PLC, in the broader context of the survival of V. vulnificus.

Glioblastoma-Specific Anticancer Activity of Pheophorbide a from the Edible Red Seaweed Grateloupia elliptica

  • Cho, MyoungLae;Park, Gab-Man;Kim, Su-Nam;Amna, Touseef;Lee, Seokjoon;Shin, Woon-Seob
    • Journal of Microbiology and Biotechnology
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    • v.24 no.3
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    • pp.346-353
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    • 2014
  • The chlorophyll-related compound pheophorbide a (Pa) was successively purified from an edible red seaweed, Grateloupia elliptica, using silica, octadecyl silica column chromatography and reversed phase-high-performance liquid chromatography, as well as the cell cycle inhibitory and apoptotic effects of Pa being investigated in U87MG glioblastoma cells. The Pa exhibited strong anticancer effects in the absence of direct photo-irradiation against various cancer cell lines, including U87MG, SK-OV-3, and HeLa cells. Among the cancer cells, the strongest anticancer activity of Pa exhibited on U87MG cells with $IC_{50}$ values of 2.8 ${\mu}g/ml$. In addition, Pa specifically had cytostatic activity on glioblastoma cells rather than human umbilical vein endothelial cells. Analysis of the cell cycle distribution showed that Pa induced G0/G1 arrest of U87 MG cells. In addition, arrested cells induced late apoptosis and DNA degradation under dark condition. These results suggest that Pa isolated from G. elliptica is a potential glioblastoma-specific anticancer agent without side effects on normal cells.

Liquid Chromatographic Determination of Etofenprox Residues in Foods with Mass-Spectrometric Confirmation

  • Lee, Young-Deuk;Kwon, Chan-Hyeok;Kwon, Ki-Sung
    • Korean Journal of Environmental Agriculture
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    • v.30 no.4
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    • pp.432-439
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    • 2011
  • BACKGROUND: An official analytical method was developed to determine etofenprox residues in agricultural commodities using high-performance liquid chromatography (HPLC). METHODS AND RESULTS: The etofenprox residue was extracted with acetone from representative samples of five raw products which comprised rice grain, apple, mandarin, cabbage, and soybean. The extract was then serially purified by liquid-liquid partition and Florisil column chromatography. For rice and soybean samples, acetonitrile/n-hexane partition was additionally coupled to remove nonpolar lipids. Reversed phase HPLC using an octadecylsilyl column was successfully applied to separate etofenprox from co-extractives. Intact etofenprox was sensitively detected by ultraviolet absorption at 225 nm. Recovery experiment at the quantitation limit validated that the proposed method could apparently determine the etofenprox residue at 0.02 mg/kg. Mean recoveries from five crop samples fortified at three levels in triplicate were in the range of 93.6~106.4%. Relative standard deviations of the analytical method were all less than 10%, irrespective of crop types. A selected-ion monitoring LC/mass spectrometry with positive atmospheric-pressure chemical ionization was also provided to confirm the suspected residue. CONCLUSION(s): The proposed method is simple, rapid and sensitive enough to be employed in routine inspection or monitoring of agricultural products for the etofenprox residue.

Antioxidant Activities and Determination of Phenolic Acids from Leaves of Perilla frutescens

  • Lee, Jin-Hwan;Kang, Nam-Suk;Ha, Tae-Joung;Ko, Jong-Min;Han, Won-Young;Suh, Duck-Yong;Park, Ki-Hun;Baek, In-Youl
    • Journal of Applied Biological Chemistry
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    • v.49 no.1
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    • pp.11-15
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    • 2006
  • Two catecholic phenolic acids (1 and 2) obtained from 80% methanolic extract of Perilla frutescens var. frutescens leaves through chromatography showed strong DPPH (1: $IC_{50}$ = 15.5 ${\mu}M$, 2: $IC_{50}$ = 11.7 ${\mu}M$) and ABTS (1: $IC_{50}$ = 5.5 ${\mu}M$, 2: $IC_{50}$ = 4.6 ${\mu}M$) radicals scavenging abilities. Antioxidant compounds contents of 1 and 2 as determined by $C_{18}$ reversed phase HPLC coupled with diode-array detector were 2.98 and 2.26 mg/g, respectively.