• Title/Summary/Keyword: High performance Liquid Chromatography

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Genetic and Functional Analyses of the DKxanthene Biosynthetic Gene Cluster from Myxococcus stipitatus DSM 14675

  • Hyun, Hyesook;Lee, Sunjin;Lee, Jong Suk;Cho, Kyungyun
    • Journal of Microbiology and Biotechnology
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    • v.28 no.7
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    • pp.1068-1077
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    • 2018
  • DKxanthenes are a class of yellow secondary metabolites produced by myxobacterial genera Myxococcus and Stigmatella. We identified a putative 49.5 kb DKxanthene biosynthetic gene cluster from Myxococcus stipitatus DSM 14675 by genomic sequence and mutational analyses. The cluster consisted of 15 genes (MYSTI_06004-MYSTI_06018) encoding polyketide synthases, non-ribosomal peptide synthases, and proteins with unknown functions. Disruption of the genes by plasmid insertion resulted in defects in the production of yellow pigments. High-performance liquid chromatography and liquid chromatography-tandem mass spectrometry analyses indicated that the yellow pigments produced by M. stipitatus DSM 14675 might be novel DKxanthene derivatives. M. stipitatus did not require DKxanthenes for the formation of heat-resistant viable spores, unlike Myxococcus xanthus. Furthermore, DKxanthenes showed growth inhibitory activity against the fungi Aspergillus niger, Candida albicans, and Rhizopus stolonifer.

DETERMINATION OF TRIETHANOLAMINE BY HIGH PERFORMANCE LIQUID CHROMATOGRAPHY WITH POST COLUMN REACTION

  • Kim, Jin-Woo;Kim, Seung-Jung;Lee, Bo-Seaub
    • Journal of the Society of Cosmetic Scientists of Korea
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    • v.14 no.1
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    • pp.1-15
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    • 1988
  • A new method for liquid chromatography with post column reaction is suggested for the separation and quantification of tertiary amines. A mixture of triethanolamine and N-ethyl diethanolamine was separated by a strong cation exchange column, followed by spectrophtometric detection of the blue colors generated from the reaction of each amine with the Folin-Ciocalteau reagent. The tertiary amines were properly separated when an eluent of pH 9.5 containing 0.5M sodium nitrate was used. Under this condition, calibration curve of triethanolamine in 2-10mg/100ml concentration range was attained. Good results were obtained when cream and shampoo preparations containing known amount of triethanolamine were analysed according to this method. In case the sample did not contain any other interfering reducing substances, the amine was quantitatively determined by the simple reaction of the samples with Folin-Ciocalteau reagent, and the subsequent spectrophotometric measurement.

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Determination of Monocrotophos Residues by HPLC

  • Lee, Young-Deuk;Kwon, Chan-Hyeok
    • Korean Journal of Environmental Agriculture
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    • v.23 no.4
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    • pp.245-250
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    • 2004
  • An analytical method was developed to determine monocrotophos residues in apple, citrus, and soil using high-performance liquid chromatography (HPLC) with ultraviolet absorption detection. Monocrotophos was extracted with acetone from apple, citrus and moist soil samples. The extract was concentrated, added with saline water, and subjected to n-hexane washing to remove nonpolar co-extractives. Dichloromethane partition was then followed to recover monocrotophos from the aqueous phase. Silica gel column chromatography was employed to further purify the extract prior to HPLC determination. Reverse-phase HPLC using an oct-adecylsilyl column was successfully applied to separate and quantitate the monocrotophos residue in sample extracts at the wavelength of 230 nm. Overall recoveries of monocrotophos from fortified samples averaged $95.3{\pm}2.1%$ (n=6), $970{\pm}0.7%$ (n=6), and $92.8{\pm}4.3%$ (n=12) for apple, citrus, and soil, respectively. The proposed method was quite reproducible and sensitive enough to replace the troublesome gas-liquid chromatographic analysis for monocrotophos residues.

A study on the optimal HPLC condition for peptides complex analysis using mass spectrometry (질량분석기를 단백질 분석에 적용하기 위한 고성능액체크로마토그래피 최적조건 연구)

  • Kwon, Sung Won;Park, Chul Hong
    • Analytical Science and Technology
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    • v.16 no.1
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    • pp.78-81
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    • 2003
  • Peptides separation in high performance liquid chromatography (HPLC) is very important for the analysis of total proteins using mass spectrometry rather than two-dimensional polyacrylamide gel electrophoresis (2D-PAGE). In this study, we investigated the optimal HPLC condition of peptides for the use of mass fingerprinting. As a result of pursuing a combination of solvent additives for HPLC, water and acetonitile containing both 0.1% trifluoroacetic acid and 0.1% acetic acid respectively showed the most efficient resolution and sensitivity.

Isolation of Ginsenoside Rh1 and Compound K from Fermented Ginseng and Efficacy Assessment on Systemic Anaphylactic Shock

  • Kim, Mi-Soon;Kwon, Bin;Park, Myeong-Soo;Ji, Geun-Eog
    • Food Science and Biotechnology
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    • v.17 no.4
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    • pp.805-808
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    • 2008
  • Ginsenosides are responsible for the pharmacological and biological activities of ginseng. In this study, ginsenoside Rh1 and compound K were isolated and purified from fermented ginseng substrate and their anti-allergic effects were assessed in compound 48/80-induced anaphylactic shock model. The fermented ginseng substrate was extracted by methanol and ginsenoside Rh1 and compound K were efficiently purified by preparative high performance liquid chromatography (prep HPLC). Their quality and quantity were analyzed by liquid chromatography-mass spectrometer (LC-MS) and HPLC. Ginsenoside Rh1 showed better anti-allergic effects than compound K in compound 48/80-induced anaphylactic shock model. This study suggested that fermented ginseng extracts with enriched Rh1 may be utilized as a potential biomaterial of functional food for the alleviation of allergic symptoms.

Optical Resolution of Dansyl Amino Acids with Addition of Benzyl-L-Hydroxyproline Copper(II) Chelate by High Performance Liquid Chromatography

  • Sun Haing Lee;Tae Sub Oh;Sang Hyun Bak
    • Bulletin of the Korean Chemical Society
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    • v.10 no.6
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    • pp.491-495
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    • 1989
  • Resolution of enantiomers of DNS-amino acids has been achieved by a reversed phase liquid chromatography with an addition of a copper(Ⅱ) complex of N-benzyl-L-hydroxyproline to the mobile phase. N-Benzyl-L-hydroxyproline was prepared and used as a chiral ligand of copper(Ⅱ) chelate for the optical resolution. The pH and the concentration of copper(Ⅱ) chelate, organic solvent, and buffer agent in the mobile phase all affect the optical resolutions of dansyl amino acids. The elution orders between D and L-DNS-amino acids were different depending on the structure of the side chain of the amino acids. The retention mechanism for the chiral separation of the dansyl amino acids can be illustrated by the equilibrium of ligand exchange and by hydrophobic interaction with $C_{18}$ stationary phase. The chiral separation can be illustrated with cis and trans effect of the ligand exchange reaction.

Microprocessor Based Laser Induced Fluorometry I. Development of System and Application to Liquid Chromatography

  • Kim, Ha-Suck;Park, Chan-Seung;Hwang, Eui-Jin;Chol, Q.-Won
    • Bulletin of the Korean Chemical Society
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    • v.5 no.6
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    • pp.253-259
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    • 1984
  • An analytical applicability of the fluorescence detection with an optical multichannel analyzer to organic dyes was studied in this work. Continuous acquisition of spectra was possible with the use of a microcomputer. The maximum acquisition rate of a spetrum with 70-point average was about 3 seconds. Floppy discs were used to store data for later use in processing. Laser induced fluorescence detector in High Performance Liquid Chromatography was chosen for an application. Fluorescein below $10^{-15}g$ was detected satisfactorily using this system. With the help of microcomputer, three dimensional chromatograms of time-wavelength-intensity were obtained.

Column cleaning, regeneration and storage of silica-based columns (실리카 기반 컬럼의 세척, 재생 및 보관 가이드)

  • Matt James;Mark Fever
    • FOCUS: LIFE SCIENCE
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    • no.1
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    • pp.1.1-1.4
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    • 2024
  • This article provides comprehensive guidance on the maintenance, cleaning, regeneration, and storage of silica-based HPLC (High-Performance Liquid Chromatography) columns. The general considerations emphasize the importance of using in-line filters and guard cartridges to protect columns from blockage and irreversible sample adsorption. While these measures help, contamination by strongly adsorbed sample components can still occur over time, leading to an increase in back pressure, loss of efficiency, and other issues. To maximize column lifetime, especially with UHPLC (Ultra-High Performance Liquid Chromatography) columns, it is advisable to use ultra-pure solvents, freshly prepared aqueous mobile phases, and to filter all samples, standards, and mobile phases. Additionally, an in-line filter system and sample clean-up on dirty samples are recommended. However, in cases of irreversible compound adsorption or column voiding, regeneration may not be possible. The document also provides specific recommendations for column cleaning procedures, including the flushing procedures for various types of columns such as reversed phase, unbonded silica, bonded normal phase, anion exchange, cation exchange, and size exclusion columns for proteins. The flushing procedures involve using specific solvents in a series to clean and regenerate the columns. It is emphasized that the flow rate during flushing should not exceed the specified limit for the particular column, and the last solvent used should be compatible with the mobile phase. Furthermore, the article outlines the storage conditions for silica based HPLC columns, highlighting the impact of storage conditions on the column's lifetime. It is recommended to flush all buffers, salts, and ion-pairing reagents from the column before storage. The storage solvent should ideally match the one used in the initial column test chromatogram provided by the manufacturer, and column end plugs should be fitted to prevent solvent evaporation and drying out of the packing bed.

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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.

A study on deoxynivalenol production by water-saturated silical gel chromatography (물포화 Silica gel chromatography에 의한 Deoxynivalenol 생산에 관한 연구)

  • Kim, Jong-shu;Choi, Min-cheol
    • Korean Journal of Veterinary Research
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    • v.32 no.3
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    • pp.413-419
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    • 1992
  • Deoxynivalenol producing isolates of Fusarium Graminearum R 6576 was grown on rice for 25 days at 19,25 and $28^{\circ}C$. Maximum production of deoxynivalenol(DON) by Fusarium graminearum R 6575 occurred at $28^{\circ}C$ and 20 days. Maximum concentration of 940 ppm DON were obtained after 20 days at an initial moisture content of 40%. A DON derivative, 15-acetyl-DON (15-ADON), was also found at concentrations of 150~300ppm after 5~10 days. Crude culture extracts were purified by water-saturated silica gel column chromatography which selectivity extracted DON when methylene chloride was as the mobile phase. Purity of crystallized DON was verified by thin layer and high performance liquid chromatography. Also this method was advantage method or production of DON and require little organic sorbent than the other methods.

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