• Title/Summary/Keyword: reversed phase

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Application of Reversed-Phase Solid Phase Extraction for the HPLC Analysis of Microcystins in Water (다량 시료중 마이크로시스틴의 농축 및 분석)

  • Kim, Myeong-Hee;Kim, Tae-Seung;Kim, Tae-Keun;Park, Sun Ku
    • Analytical Science and Technology
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    • v.13 no.3
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    • pp.399-402
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    • 2000
  • To determine the concentrations of microcystins present in lake water or in tap water using high performance liquid chromatography, it is necessary to concentrate a large volume of water samples (about 20 L) into very small volume (0.1-0.3 mL). Concentration can be conveniently done when disc type solid phase extraction (SPE) apparatus is used. Using this apparatus we have investigated the recovery rates of three kinds of microcystins, RR, YR, LR. The recovery rates were relatively low and the reproducibilities were not good either. It is expected, however, that the appropriate selection of the disc conditioning and eluting solvents and reproducible reconcentration process after SPE will improve both the recovery rates and the reproducibilities.

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Tensile Properties of High Mn Austenitic Stainless Steel with Two Phases of Martensite and Austenite (마르텐사이트와 오스테나이트의 2상 조직을 갖는 고 Mn 오스테나이트계 스테인리스강의 인장성질)

  • Kim, Young-Hwa;Kang, Chang-Yong
    • Journal of Ocean Engineering and Technology
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    • v.27 no.4
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    • pp.9-13
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    • 2013
  • The tensile properties of high manganese austenitic stainless steel with the two phase structures of deformation-induced martensite and reversed austenite were studied. Reversed austenite with an ultra-fine grain size of less than $0.3{\mu}m$ was obtained by reversion treatment. The two phases structures of deformation-induced martensite and reversed austenite were obtained by an annealing treatment in the range of $500^{\circ}C-700^{\circ}C$ for various times in 70% cold- rolled high-manganese austenitic stainless steel. The volume fraction of the reversed austenite increased rapidly with increases in the annealing temperature and time. In the stainless steel with the two phases of austenite and martensite, the strength decreased rapidly, while the elongation increased slowly and then rapidly increased with an increase in the volume fraction of the reversed austenite. Therefore, the strength and elongation were strongly controlled by the volume fraction of reversed austenite. A good combination of high strength and elongation could be obtained by the mixed structure of reversed austenite and deformation-induced martensite.

Separation of Functionalized Heterocyclic Compounds by High Performance Liquid Chromatography(I) (고성능 액체 크로마토그래피에 의한 기능성 헤테로고리화합물의 분리(I))

  • Lee, Kwang-PilI;Cho, Yun Jin;Lee, Young Cheol
    • Analytical Science and Technology
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    • v.10 no.6
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    • pp.408-417
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    • 1997
  • Normal phase or reversed phase liquid chromatographic separation of some structural isomers of functionalized heterocyclic compounds has been carried out by using several different columns and various mobile phases. The optimal experimental conditions for separation of structural isomers were found on a ternary solvent system including alcohol as a modifier. This polar modifier is preferentially adsorbed onto strong adsorption site, leaving a more uniform population of weaker site that then serve to retain the sample. This 'deactivation' of the adsorbent leads to a number of improvements in subsequent separations. The optimal mobile phase system of separation were found on normal phase on structural isomers. Retention mechanism of normal phase system was also studied depending on adsorption strength between solute and stationary phase of column. However, retention factors of reversed phase system were found on hydrophobic interaction with solvophobic effect.

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Determination Method of Puerarin and Daidzin from Puerariae Radix by Reversed-Phase HPLC with Pulsed Amperometric Detection (RP-HPLC-PAD를 이용한 갈근(葛根)과 갈근(葛根) 함유 처방의 Puerarin과 Daidzin 분석)

  • Chiao, Chen-Ying;Kwon, Ha-Jeong;Jeong, Ji-Seon;Lee, Je-Hyun;Hong, Seon-Pyo
    • The Korea Journal of Herbology
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    • v.23 no.4
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    • pp.171-177
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    • 2008
  • Objectives: This study presents a reversed-phase high-performance liquid chromatography- pulsed amperometric detection(RP-HPLC-PAD) method for the determination of puerarin and daidzin in Puerariae Radix extract and Chinese medicinal preparations. Methods: Chromatographic separation was performed using a 10% acetonitrile with a reversed-phase column(Unison UK-C18, $100mm{\times}2.0mm$ I.D.; $3{\mu}m$). The analyses were detected by pulsed amperometric detector(PAD) in alkaline conditions by combining with post-column NaOH solution. Geniposide was used as an internal standard. Results: The limit of detection(S/N=3) and the limit of quantification(S/N=10) were 0.025 ng, 0.075 ng for puerarin, and 0.05 ng, 0.15 ng for daidzin, respectively. The intra- and inter-day precisions(RSDs) were less than 6.5% and average recoveries of puerarin were 99.7-101.3% and those of daidzin were 101.0-102.8%. Conclusions: According to above results, we developed a determination method for puerarin and daidzin in Puerariae Radix with high sensitivity and selectivitely.

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Determination of Uranium and Thorium in Natural Ground Water by Reversed Phase Liquid Chromatography (역상액체크로마토그래피에 의한 지하수 중 U 및 Th의 분리정량)

  • Lee, Chang Heon;Jo, Gi Su;Seo, Mu Yeol;Lee, Won
    • Journal of the Korean Chemical Society
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    • v.38 no.7
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    • pp.502-508
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    • 1994
  • The measurement of trace amounts of uranium(VI) and thorium(VI) in the solutions containing high concentration of dissolved salts was carried out. The procedure using reversed phase liquid chromatography with trace enrichment techniques has been developed to cope with the high salt content of the samples. 2 ml of sample were passed through a small C_{18}$ reversed phase enrichment column with ${\alpha}$-HIBA eluent (0.11 M, pH 5.5) where the uranium and thorium were separated from other constituents and concentrated. The uranium and thorium were then backflushed from the column onto a deactivated C_{18}$ reversed phase analytical column where furthe separation was achieved with a mixed eluent (pH3.0, 0.17M ${\alpha}$-HIBA/0.0038 M 1-pentanesulfonate). The separated species were determined spectrophotometrically by postcolumn reaction with Arsenazo III, the chromogenic reagent. Detection limits were found within 1 ppb range for both species.

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Mass Transfer of Lysozyme Extraction Using Reversed Micelles (역미셀을 이용한 Lysozyme 추출에 대한 물질 전달)

  • 전병수;김석규;윤성옥;송승구
    • KSBB Journal
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    • v.16 no.3
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    • pp.241-245
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    • 2001
  • Mass transfer rates have been measured for the extraction of enzyme from aqueous solution into a reverse-micelle phase at $25^{\circ}C$. The 420 mL vessel was carefully designed to maintain a planar interface between the aqueous and solvent phases, so allowing precise measurement of interfacial area, has been investigated. Sodium di-2-ethylhexyl sulfosuccinate(AOT) was the surfactant used. Factors varied included: agitator speed, pH, ionic strength and surfactant concentration. Samples were taken from the solvent phase at 15min intervals, and the amount of enzyme extracted was measured by UV absorption at 280 nm. The observed Sherwood numbers for the aqueous phase $Sh_1$were correlated interms of the aqueous phase Reynolds number $Re_1$, and modified Schmidt number $Sc_1$. $Sh_1=0.664Re_1^{0.5}Sc_1^{0.33}$

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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.

Enantiomeric Purity Test of Bevantolol by Reversed-Phase High Performance Liquid Chromatography after Derivatization with 2,3,4,6-tetra-O-acetyl-$\beta$-D-glucopyranosyl Isothiocyanate

  • Kim, Kyeong-Ho;Heo, Sung-Young;Hong, Seon-Pyo;Lee, Beom-Chan
    • Archives of Pharmacal Research
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    • v.23 no.6
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    • pp.568-573
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    • 2000
  • A reversed-phase high-performance liquid chromatographic method was developed to determine the optical purity of bevantolol enantiomers. (S)-(-)-Menthyl chloroformate((-)-MCF), (S)-(-)-$\alpha$-methylbenzyl isocyanate((-)-MBIC) and 2,3,4,6-tetra-O-acetyl-$\beta$-D-glucopyranosyl isothiocyanate(GITC), which can react with the secondary amine group of bevantolol were investigated as chiral derivatization reagents. Among them indirect chiral HPLC method using CITC gave the best result. The derivatization proceeded quantitatively within 20 min at room temperature. Separation of the enantiomers as diastereomers was achieved by reversed-phase HPLC within 20min using ODS column. Different ratios of (S)-(-)-bevantolol and (R)-(+)-bevantolol were prepared. Enantiomeric separation of these mixtures took place on a chiralcel OD column or, after derivatization with GITC, on a ODS column. No racemization was found during the experiment. This method allowed determination of 0.05% of either enantiomer in the presence of its stereoisomer and method validation showed adequete linearity over the required range.

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Isolation of Phytochemicals from Salvia plebeia Using Countercurrent Chromatography Coupled with Reversed-phase HPLC

  • Kil, Hyun Woo;Rho, Taewoong;Seo, Young Ju;Yu, Aram;Yoon, Kee Dong
    • Natural Product Sciences
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    • v.26 no.3
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    • pp.236-243
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    • 2020
  • Salvia plebeia R. Br. is a plant which has been used as an edible crop and traditional medicine in Asian countries. In this study, HPLC-PDA analysis and countercurrent chromatography (CCC) coupled with reversed-phase (RP) HPLC method were applied to isolate ten isolates from 3.3 g of n-butanol soluble extract from hot-water extract of S. plebeia. The use of CCC enabled us to efficiently fractionate the starting material with less sample loss and facilitate the isolation of compounds from S. plebeia extract using RP-HPLC. The isolates were determined to be caffeic acid (1), 6-hydroxyluteolin 7-O-β-D-glucoside (2), eudebeiolide B (3), (R)-rosmarinic acid (4), homoplantaginin (5), eudebeiolide D (6), plebeiolide C (7), salpleflavone (8), eupafolin (9) and hispidulin (10) based on the spectroscopic evidence.

Analysis of amino acids in the liver of Hamster treated with Dimethylnitrosamine (Dimethylnitrosamine이 투여된 Hamster 간 속의 아미노산 분석)

  • Kim, Soo Gyung;Jung, Ha Seung;Park, Taek Kyu
    • Analytical Science and Technology
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    • v.7 no.2
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    • pp.165-171
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    • 1994
  • The effect of the chemical carcinogen dimetylnitrosamine(DMN) on the composition of amino acids of the liver in hamsters orally administered with DMN was studied by using the reversed phase high performance liquid chromatography technique. In the liver, the concentration of aspartic acid, glycine, glutamine, histidine, proline, tyrosine and leucine were increased ca. 2-fold of those observed in liver of control group, valine and tryptophan were increased ca. 3-fold, phenylalanine was markedly increased ca. 10-fold, whereas the concentration of threonine was decreased, serine, alanine, arginine, methionine, isoleucine and lysine were unchanged, respectively.

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