• Title/Summary/Keyword: Reversed-Phase HPLC

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Effect of Eluent Electrolyte on the Retention Behavior of Structural Isomers of Phenols in HPLC. (HPLC 에서 페놀류의 구조 이성질체의 머무름 거동에 대한 전해질 용리액의 효과)

  • Lee, Seon Haeng;O, Dae Seop;Park, Gi Ho
    • Journal of the Korean Chemical Society
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    • v.34 no.1
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    • pp.44-50
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    • 1990
  • The liquid chromatographic retention behavior of structural isomers of phenols was investigated by a change of the mobile phase properties. The retention behavior of structural isomer of phenols in reversed phase liquid chromatography was affected by eluent electrolyte added. It can be seen that this behavior is illustrated by a mechanism of Langmuir isotherm and ion exchange between phenolate and the reversed phase coated with ions. The retention behavior was represented as two different areas according to the concentration of the electrolytes. These areas can be explained as counter ion and co-ion effect, respectively. The maximum retention values were dependent not upon the kinds of organic modifier but upon the kinds of electrolyte.

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Determination of Nebramycin Factor 2,4,5,5',6 and Kanamycin A in Fermentation Broth of Streptoalloteichus hindustanus ATCC 31218 Mutant Using 2,4-Dinitrofluorobenzene(DNFB) as a Derivatizing Agent by High Performance Liquid Chromatography (HPLC법에 의한 2,4-디니트로플루오로벤젠을 유도체화제로 한 Streptoalloteichus hindustanus ATCC 31218 변이균의 배양액 중 네브라마이신 펙터 2,4,5,5',6, 가나마이신 A 분석)

  • 박영근;박명용;김승철;양호길
    • YAKHAK HOEJI
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    • v.37 no.1
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    • pp.1-8
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    • 1993
  • A procedure for the high-performance liquid chromatographic determination of Nebramycin factors in fermentation broth of Streptoalloteichus hindustanus ATCC 31218 mutant was investigated using pre-column derivatization and LTV detection. The method is based on pre-column derivatization of Nebramycin factors with 2,4-dinitrofluorobenzene(DNFB) in the presence of Tris (hydroxymethyl)aminoethane. The chromatographic separation of derivatives of Nebramycin factors and unknown impurities is achieved using reversed-phase column (NOVA-PAK $C_{18}$, Waters Co.) and AcCN : H$_{2}$O : AcOH (53.0:46.5:0.5) as a mobile phase. The mixture of these derivatives were separated within 35 minutes and the optimum wavelength($\lambda_{max}$ ) of the UV detector was 353 nm. The linearity of response for derivatives of Nebramycin factors is demonstrated for concentrations up to 500 $\mu\textrm{g}$/ml and the relative standard deviation is less than 0.79%. Detection limit was 1.67 ng for the 10 $\mu\textrm{l}$ sample volume employed.

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Identification of Proteins in Egg White Using Ion Exchange Cartridge and RP-HPLC (이온교환 카트리지와 RP-HPLC를 이용한 난백 단백질의 확인)

  • Kim, Hyun Moon;Kim, Ah Reum;Lee, Chang Soo;Kim, In Ho
    • Korean Chemical Engineering Research
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    • v.50 no.4
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    • pp.713-717
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    • 2012
  • Approximately forty proteins in egg white have been widely studied for their functional properties. To develop a procedure of separation for pure and non-altered proteins from egg white, purification study was conducted to isolate lysozyme, ovotransferrin, and ovalbumin. Ion exchange cartridge can selectively separate proteins from egg white, and reversed-phase HPLC (RP-HPLC) could identify separated proteins. Proteins in egg white were purified by HI trap ion exchange cartridge SP and Q with buffers pH 8.0 and 5.2. C18 column (Phenomenex, USA) was used for RP-HPLC analysis and isocratic mobile phase was used with acetonitrile (ACN)/distilled water (DW)/trifluoroacetic acid (TFA) in the ratio of 50/50/0.1. Comparing the retention times of standards in RP-HPLC experiments showed that ovotransferrin, ovalbumin, and lysozyme in egg white were eluted successively in the RP-HPLC column after the pretreatment in SP and Q ion exchange cartridges.

Validations of Analysis Methods for Decursin and Decursinol Angelate of Angelicae gigantis Radix by Reversed-Phase Liquid Chromatography

  • Lee, Jong-Pill;Chang, Seung-Yeup;Park, Sang-Yong
    • Natural Product Sciences
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    • v.10 no.6
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    • pp.262-267
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    • 2004
  • A reversed-phase liquid chromatographic method for decursin and decursinol angelate of Angelicae gigantis Radix, an important crude drug in Korean traditional medicine, was developed and validated. Decursin and decursinol angelate, the structure isomer (pyranocoumarin) each other, are the main organic constituents in Angelicae gigantis Radix. This method was developed using a RP-18 column, UV detector at 280 nm and 50% acetonitrile solution containing 0.01 M sodium dodecyl sulfate and 25 mM sodium dihydrogen phosphate (pH 5.0) as the mobile phase. Various validation parameters were included and evaluated satisfactorily. Linearity was established in range 2-75 mg/ml of decursin and decursinol angelate (correlation coefficient = 0.9997 and 0.9995, respectively). This analytical method showed good accuracy (98.1% and 99.5%, respectively). Precision (repeatability) revealed a relative standard deviation value of 1.71% (decursin) and 3.19% (decursinol angelate). For intermediate precision measure the considered variables were equipment and days. A robustness test showing the influence of deferent counter-ion concentration in mobile phase was also performed.

Simultaneous HPLC Analysis of Arachidonic Acid Metabolites in Biological Samples with Simple Solid Phase Extraction

  • Kim, Hyung-Gun;Huh, Young-Na;Park, Kun-Suk
    • The Korean Journal of Physiology and Pharmacology
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    • v.2 no.6
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    • pp.779-786
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    • 1998
  • A reversed-phase high-performance liquid chromatogrphy (RP-HPLC) has been developed to analyze the metabolites of arachidonic acid based on the specificities of ultraviolet absorption of these various metabolites and is sensitive to the nanogram level. This procedure makes it possible to extract complex mixtures of eicosanoids efficiently with a single step and to analyze them simultaneously by RP-HPLC from biological samples using octadesylsilyl silica extraction column and $PGB_2$ as an internal standard. The cyclooxygenase products {prostaglandin $(PG)D_2,\;PGE_1,\;PGE_2,\;PGF_{1{\alpha}},\;PGF{2{\alpha}},\;6-keto-PGF_{1{\alpha}},$ and thromboxane $B_2(TXB_2)}$ and lipid peroxidation product, isoprostanes, of arachidonic acid were monitored by one isocratic HPLC system at 195 nm wavelength. The lipoxygenase products ${leukotriene(LT)B_4,\;LTC_4,\;LTD_4,$ and 5-hydroxyeicosatetraenoic acid (5-HETE), 12-HETE, 15-HETE} were measured by another isocratic HPLC system at 280 nm for LTs and 235 nm for HETEs. This method provides a simple and reliable way to extract and assess quantitatively the final arachidonic acid metabolites.

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Simultaneous determination of sunscreen agents in cosmetics by HPLC (HPLC를 이용한 화장품 중의 자외선 차단제 동시분석방법 연구)

  • Lee, Yong-Hwa;Yang, Jae-Chan
    • Journal of the Korean Applied Science and Technology
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    • v.29 no.4
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    • pp.577-584
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    • 2012
  • The simultaneous analysis of sun screen agents in commercial cosmetic samples was carried out by High Perfomance Liquid Chromatography(HPLC). The cosmetic samples are directly dissolved in Tetrahydrofurane(THF) and filtered using $0.45{\mu}m$ filter. The water/methanol/THF was used for the mobile phase of gradient conditions. An Extend C18 reversed-phase column and the selected UV/Visible detector was applied. The analysis results of HPLC showed good linearity with correlation coefficient of $r^2$=0.9992 in the rage of $50{\sim}800{\mu}g/mL$ and detection limit of $0.01{\mu}g/mL$.

Prediction of Retention Behavior of Alkyl Benzenes by Hydrophobicity Parameters in Reversed-Phase Column (소수성 파라메터를 적용한 알킬벤젠류의 역상컬럼내의 용출거동 예측)

  • Lee, Chang-Young;Park, Myung-Yong;Lee, Yong-Moon
    • YAKHAK HOEJI
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    • v.53 no.5
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    • pp.281-285
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    • 2009
  • The retention of solutes in reversed-phase high-performance liquid chromatography depends on their hydrophobicity. Although the retention behaviors of alkyl benzenes have been reported so far, quite a few authors have mentioned the retention behavior of alkyl benzenes with plural hydrophobicity parameters. In this sense, we were interested in the retention behaviors of alkyl benzenes having benzene moiety and increasing alkyl chain. In this study, we therefore investigated the retention behavior of alkyl benzenes in reversed-phase high-performance liquid chromatography in order to obtain information concerning the effects of the aromatic moiety and the carbon chain on the retention mechanism by comparing their capacity factor (k') in relation to the carbon chain length. The eluent acetonitrile ($CH_3CN$) showed high selectivity on alkyl benzenes, showing the high difference of capacity factor (${\Delta}log\;k'$) between toluene and octyl benzene. Indeed, the ${\Delta}log\;k'$ of 80% $CH_3CN$ represented 1.42- and 4.25-times longer than 90% MeOH and 60% THF, respectively. The hydrophobicity parameters, van der Waals volume, bond constant, partition constant, $\pi$-energy effect and enthalpy were evaluated with the capacity factor (k') of alkyl benzenes eluted on 80% CH3CN, 90% MeOH and 60% THF, respectively. The best eluent for predicting retention behavior of alkyl benzenes was 90% MeOH ($R^2$ 0.999). The three parameters, van der Waals volume, bond constant and partition constant were well coincident to log k' by increasing alkyl benzenes. However, $\pi$-energy effect and enthalpy were severely disagreeable. Taken together, van der Waals volume, bond constant and partition constant were a reliable parameters to predict the retention behaviors of alkyl benzenes on reversed-phase column.

Chiral separation of $\beta$-blockers after derivatizalion wilh a new chiral derivalizalion agen1.GATC and Comparison wilh GITC on derivalizing conditions and chromalographic paramelers

  • Ko, Mi-Young;Shin, Dae-Hong;Seo, Sang-Hun;Kim, Kyeong-Ho
    • Proceedings of the PSK Conference
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    • 2002.10a
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    • pp.398.1-398.1
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    • 2002
  • A new chiral derivatization agent with sugar moiety. 2, 3.4, 6-tetra-O-acetyl-D-galactopyranosyl isothiocyanate (GATC) was synthesized. Several $\beta$-blockers were investigated for the possible separation of the enantiomers by reversed-phase HPLC after derivatization with this new chiral derivatization agent (GATC). GATC was reacted readily with $\beta$-blockers at room temperature and the reaction mixture could directly be injected into the HPLC system. (omitted)

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Synthesis, Characterization and Enhanced Selectivity in RP-HPLC of Polar Carbonyl Group Embedded Poly (Vinyl Octadecanoate) Grafted Stationary Phase by Simple Heterogeneous "Graft from" Technique

  • Rana, Ashequl A.;Karim, Mohammad Mainul;Takafuji, Makoto
    • Bulletin of the Korean Chemical Society
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    • v.32 no.1
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    • pp.77-82
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    • 2011
  • A new high performance liquid chromatograpgy (HPLC) stationary phase that possesses an internal carbonyl functional group is synthesized by heterogeneous "graft from" method. This new stationary phase, poly (vinyl octadecanoate) grafted silica (Sil-2) is then characterized by different physico-chemical methods such as diffuse reflectance infrared fourier transform, suspension state $^1H$ NMR, solid state $^{13}C$ CP/MAS NMR, $^{29}Si$ CP/MAS NMR, elemental analysis and thermogravimetric analysis. Chromatographic properties of Sil-2 were evaluated under reversed phase condition by separating polycyclic aromatic hydrocarbons (PAHs) and comparing the chromatographic results with those on polymeric as well as monomeric octadecylated silica stationary phases.

Solid-Phase Extraction of Sulfamerazine from Shrimp Residue and Determination by Reversed Phase High Performance Liquid Chromatography

  • Jang, Won-Cheoul;Heo, Gang-Joon
    • Toxicological Research
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    • v.12 no.2
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    • pp.163-169
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    • 1996
  • The focus of this study was to investigate the suitable analytical methods for measurement of sulfamerazine and its metabolite in shrimp hepatopancreas and tail tissue, in addition to the methods for the optimization of solid-phase extraction cartridge conditions and the elucidation of sulfamerazine concentrations in aqueous buffer using HPLC with UV and EC detectors. Compared with UV detector the EC detector appears to be 10 times more sensitive than that of the UV detector. After the shrimp was exposed to 10 ppm sulfamerazine, the accumulation levels of sulfamerazine and its metabolite in tail tissue, which is edible portion, were considerably lower than 0.1 ppm. The data indicate that sulfamerazine continues to be a candidate for use at levels of sulfamerazine concentration used in aquaculture of shrimp.

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