• 제목/요약/키워드: Reversed-Phase HPLC

검색결과 357건 처리시간 0.032초

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

  • 이선행;오대섭;박기호
    • 대한화학회지
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    • 제34권1호
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    • pp.44-50
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    • 1990
  • 페놀류의 구조 이성질체의 액체 크로마토그래피적 머무름거동을 이동상의 성질을 변화시켜 조사했다. 역상 액체 크로마토그래피에서 페놀류의 구조이성질체에 대한 머무름거동은 첨가한 용리액의 전해질에 영향을 받았다. 이 머무름거동은 Langmuir의 흡착등온선과 페놀이온과 전해질 이온으로 코우팅 된 정지상 사이의 이온교환과정으로 설명할 수 있다. 이 거동은 전해질의 농도에 따라 두 영역으로 나누어져서 반대이온효과가 나타나는 부분과 동종이온효과가 나타나는 부분으로 설명할 수 있다. 머무름값이 극대가 나타나는 것은 유기용매의 종류에는 관계없이 전해질의 종류와 농도에 따라 다르게 나타났다.

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HPLC법에 의한 2,4-디니트로플루오로벤젠을 유도체화제로 한 Streptoalloteichus hindustanus ATCC 31218 변이균의 배양액 중 네브라마이신 펙터 2,4,5,5',6, 가나마이신 A 분석 (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)

  • 박영근;박명용;김승철;양호길
    • 약학회지
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    • 제37권1호
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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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이온교환 카트리지와 RP-HPLC를 이용한 난백 단백질의 확인 (Identification of Proteins in Egg White Using Ion Exchange Cartridge and RP-HPLC)

  • 김현문;김아름;이창수;김인호
    • Korean Chemical Engineering Research
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    • 제50권4호
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    • pp.713-717
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    • 2012
  • 난백에 있는 40여 가지의 단백질의 기능적 특성에 대한 연구가 폭 넓게 진행되고 있지만, 그 특성이 모두 확인되지 않았다. 난백에서 순수하면서 변성되지 않은 단백질을 분리하는 방법을 연구하기 위해 본 연구에서는 난백 중 함량이 많은 lysozyme, ovotransferrin, ovalbumin을 분리했다. 난백에서 단백질을 다른 분리 방법보다 선택적으로 분리가 가능한 이온 교환 크로마토그래피를 이용했다. 그리고 분리된 단백질을 동정하기 위해서 Reversed-phase HPLC (RP-HPLC)를 사용했다. 이온 교환 크로마토그래피에서 HI Trap ion exchange cartridge (GE Healthcare, USA)를 사용했고, 그 중에서 양이온 교환 수지 SP (완충용액 pH 8.0, pH 5.2)와 음이온 교환 수지 Q (완충용액 pH 8.0)로 난백 단백질을 분리하였다. 분리한 난백 단백질들의 동정용 RP-HPLC에서는 C18 컬럼(Phenomenex, USA)을 사용했고 등용매 용리에서 이동상으로 acetonitrile (ACN)/distilled water (DW)/trifluoroacetic acid (TFA)의 부피비를 50/50/0.1로 사용했다. 이온 교환 크로마토그래피에 의해 각 단계에서 분리된 용출용액을 RP-HPLC으로 분석한 크로마토그램과 표준시료의 크로마토그램의 체류시간을 비교하여 난백에 포함된 ovotransferrin, ovalbumin, lysozyme이 순차적으로 용출됨을 알았다.

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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    • 제10권6호
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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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    • 제2권6호
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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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HPLC를 이용한 화장품 중의 자외선 차단제 동시분석방법 연구 (Simultaneous determination of sunscreen agents in cosmetics by HPLC)

  • 이용화;양재찬
    • 한국응용과학기술학회지
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    • 제29권4호
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    • pp.577-584
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    • 2012
  • 액체크로마토그래피법(HPLC)을 이용하여 화장품에 들어있는 자외선차단제류 등을 동시 분석하였다. 화장품 시료를 Tetrahydrofurane(THF)에 직접 용해시키고 $0.45{\mu}m$ 필터로 여과하여 물/메탄올/THF를 이동상으로 하여 Extend C18의 비극성 컬럼을 사용하여 기울기용리조건에서 20분 안에 분리하여 UV/Vis detector방법으로 정량하였다. HPLC분석결과 검량선은 $50{\sim}800{\mu}g/mL$ 농도범위에서 상관계수가 $r^2$=0.9992 이상의 좋은 직선성을 나타내었으며 검출한계는 $0.01{\mu}g/mL$였다.

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

  • 이창영;박명용;이용문
    • 약학회지
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    • 제53권5호
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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
    • 대한약학회:학술대회논문집
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    • 대한약학회 2002년도 Proceedings of the Convention of the Pharmaceutical Society of Korea Vol.2
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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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    • 제32권1호
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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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    • 제12권2호
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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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