• Title/Summary/Keyword: reverse phase high-performance liquid chromatography

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Liquid Chromatographic Separation of Salicylic Acid and Its Derivatives Using Amberlite XAD-Copolymers (Amberlite XAD-공중합체를 이용한 살리실산 및 크로마토그래피적 분리 그 유도체들의 액체)

  • Yong Soon Chung;Taik Hyuk Lee;Young Ja Moon;Dai Woon Lee
    • Journal of the Korean Chemical Society
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    • v.33 no.1
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    • pp.70-81
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    • 1989
  • Reverse phase liquid chromatographic elution behaviors of salicylic acid and its derivatives were studied with classical and modern high-performance liquid chromatography(HPLC) using Amberlite XAD-4 and XAD-7 resin packed columns. Capacity factors(k') were determined in the comparatively high concentration(from 0. 010F to 0. 150F) of ferric nitrate-50% methanol solution to elucidate the elution behaviors with classical method. On the other hand, k's were measured in the various concentrations of methanol and ferric nitrate(from $2.5{\times}10^{-4}F\;to\;1.0{\times}10^{-3}F$) solution of pH 2. 25 and 293K with HPLC to explain the elution mechanism of them, and to find their optimum separation condition. As a result, it was found that log k's of salicylic acid and its derivatives were decreased with increasing the concentration of ferric ion, and the decreasing slopes of the compounds on the increasing ferric ion concentrations were related with each stability constant of ferric salicylates. Some isomers of derivatives of salicylic acid could be separated in the optimum condition.

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Bitter Peptides Derived from ${\alpha}_{s1}-and\;{\beta}-Casein$ Digested with Alkaline Protease from Bacillus subtilis (Bacillus subtilis의 염기성 프로테아제로 분해된 ${\alpha}_{s1}$- 및 ${\beta}$-카세인에서 분리된 쓴 맛 펩타이드)

  • Sohn, Kyung-Hyun;Lee, Hyong-Joo
    • Korean Journal of Food Science and Technology
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    • v.20 no.5
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    • pp.659-665
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    • 1988
  • The ${\alpha}_{s1}$-and ${\beta}$-casein were purified by DEAE-cellulose chromatography and digested with alkaline protease from Bacillus subtilis. Bitter fractions from the hydrolyzates were isolated using n-butanol extraction, Sephadex G-25 gel chromatography, and high performance liquid chromatography. Peptide mixtures were separated by reverse-phase octadecyl silica column with linear gradient of 0-80% acetonitrile containing 0.1% trifluoroacetic acid. Major peaks were combined from replicate chromatographies and the bitterness of each peak was evaluated. The bitter-tasting peaks were rechromatograpied until isolated peaks were obtained. Three different bitter peptides(BP-I, BP-II, BP-III) were obtained from the ${\alpha}_{s1}$-casein hydrolyzate. BP-I was eluted at 34% acetonitrile and BP-II, 35%, BP-III, 26%, respectively. Two bitter peptides(BP-IV, BP-V) were isolated from the ${\beta}-casein$ hydrolyzate: BP-IV was eluted at 40% acetonitrile and BP-V, 42%. BP-V was the most hydrophobic peptide in the five bitter peptides. However, BP-I and BP-II tasted more bitter than BP-IV and BP-V.

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An analytical method of soap biodegradability with fatty acid p-BPB derivatives (지방산의 p-BPB 유도체에 의한 비누 생분해도 분석 방법)

  • Oh, Se-Woong;Lee, Ja-Kyoung;Chung, Yong;Chang, Sug-Youn;Kim, Yeo-Kyung
    • Analytical Science and Technology
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    • v.6 no.1
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    • pp.9-19
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    • 1993
  • Fatty acid salts derived from soap can be transferred into a typical derivative with p-bromophenacyl bromide using crown ether, a catalizer by the solid-liquid phase transfer reaction in nonpolar, aprotic solvents and separated by the reverse phase high performance liquid Chromatography (RP-HPLC) and determined using UV detector. The minimal limit of detection was defined at approximately 10~50ng in accordance with the chain length. The derivatization reaction in the presence of EDTA can be applied mot only to the calcium salts but also to the other various metal salts. The recoveries of fatty acid derivatizations in the absence and presence of the midium containing the yeast extract were obtained $95.4{\pm}1.2$, and $85.2{\pm}2.4%$ respectively. The analytical method would be applicable to determine the biodegradation of fatty acid salts in nature as well as in artificial condition such as shaker flask-medium method.

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Determination of Bisphenols Migrating from Epoxy Can Coatings to Aqueous Food Simulants (통조림관 코팅제에서 식품유사용매로 이행되는 비스페놀류의 분석)

  • Kang, Kyung-Mo;Shin, Hyo-Sun
    • Korean Journal of Food Science and Technology
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    • v.32 no.3
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    • pp.570-577
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    • 2000
  • A method was presented for determination of bisphenol F(BPF), bisphenol A(BPA), bisphenol F diglycidyl ether(BFDGE) and bisphenol A diglycidyl ether(BADGE) in 3 aqueous-based food simulants (water, 4% acetic acid, 20% ethanol) by reverse-phase high performance liquid chromatography(RP-HPLC)with fluorescence detection and gas chromatography with mass selective detection(GC/MSD). All the calibration lines in the range of $5{\sim}800\;{\mu}g/L$ had correlation coefficients greater than 0.9998 and detection limits of less than $1.2\;{\mu}g$ bisphenols/L. Precision at $200\;{\mu}g/L$ was under 3.1%. Recoveries of bisphenols simultaneously spiked to aqueous food simulants exceeded 95% for BPF and BPA but about 80% for BFDGE and BADGE. However, recoveris of BFDGE and BPADGE respectively spiked increased upto 95%. Detection limits in recovery test were less than $0.40\;{\mu}g$ bisphenols/L. In migration test bisphenols were determined by RP-HPLC coupled with confirmation by GC/MSD. Can coatings of epoxy phenol, modified epoxy, epoxy ester phenol and thermoset vinyl were exposed to the 3 aqueous food simulants. BPF, BFDGE and BADGE were not detected in all the can coatings but BPA was detected in 4% acetic acid and 20% ethanol in all the can coatings except modified epoxy.

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Optimization of HPLC Method and Clean-up Process for Simultaneous and Systematic Analysis of Synthetic Color Additives in Foods (식품 중 타르색소의 동시분석 및 계통분석을 위한 HPLC 분석조건 및 정제과정 확립)

  • Park, Sung-Kwan;Hong, Yeun;Jung, Yong-Hyun;Lee, Chang-Hee;Yoon, Hae-Jung;Kim, So-Hee;Lee, Jong-Ok
    • Korean Journal of Food Science and Technology
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    • v.33 no.1
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    • pp.33-39
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    • 2001
  • To develop a method for separation process using Sep-pak $C_18$, simultaneous and systematic analysis of 8 permitted and 11 non-permitted synthetic food colors in Korea, optimization of analysis conditions for reverse phase ion-pair high performance liquid chromatography was carried out. For the best result of Sep-pak $C_18$ separation the pH of color standard mixture solution was $5{\sim}6$ and 0.1% HCl-methanol solution were set as eluent. The colors eluated from Sep-pak $C_18$ cartridge were determined and confirmed by high performance liquid chromatography with a photodiode array detector at 420 nm for yellow colors type, at 520 nm for red colors type, at 600 nm for blue and green colors type and at 254 nm for mixed colors. Conditions for HPLC analysis were as follows: column, Symmetry $C_18$ (5 m, 3.9 mm $i.d.{\times}150\;mm$); mobile phase, 0.025 M ammonium acetate (containing 0.01 M tetrabutylammonium bromide) : acetonitrile : methanol (65 : 25 : 10) and 0.025 M ammonium acetate(containing 0.01 M tetrabutylammonium bromide) : acetonitrile : methanol (40 : 50 : 10); flow rate, 1 mL/min. It takes 35 minutes for simultaneaus analysis and 18 minutes for systematic analysis. The detection limits range of each colors were $0.01{\sim}0.05\;{\mu}g/g$.

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A Study on Qualitative and Quantitative Analysis of Major ingredients in Scutellariae radix (황금(Scutellariae radix)의 주요 성분의 정성 및 정량분석에 관한 연구)

  • Rhee, Jae-Seong;Woo, Eun Ran;Kim, Nam-Hyuk;Lee, Eun-Ju;An, Duk-Kyun;Lee, Je-Hyun;Park, Seong Kyu;Park, Ho-Koon
    • Analytical Science and Technology
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    • v.10 no.2
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    • pp.91-104
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    • 1997
  • Scutellariae radix has been used on the control of body fever as oriental medicine for thousand years. Analytical aspect for the main components of Scutellariae radix was set up improving sensitivity and resolution. The analysis of 3 different flavonoids present in Scutellariae radix-baicalin, baicalein, wogonin-was conducted by means of high performance liquid chromatography with ODS reverse phase column in conjunction with a Photo Diode Array UV detector(280nm) at $40^{\circ}C$. Mobile phase was carried out at 1mL/min, composed of acetonitrile and 0.1M phosphoric acid in the form of a gradient method. Under these circumstances the retention time for baicalin, baicalein, wogonin was 7.65, 11.65 and 14.12 minutes respectively. As a result for the efficiency on extraction of active ingredients with proposed analytical process according to it's growing districts, Sunchang in Junbuk for baicalin and Bulkyo in, Junnam for bicalein and wogonin have shown the best results. Even the extraction at room temperature was satisfactory. Among acids, 0.1M acetic acid revealed the best achievements. The mixture of acetonitrile and 0.2M phosphoric acid(75:25) has been shown the best efficiency as well as stability for the extraction of active ingredients.

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The Optimum Conditions for the Simultaneous Determination of Neurotransmitters in Rat Brain Striatum by High Performance Liquid Chromatography with Electrochemical Detection (HPLC-ECD를 이용한 흰쥐 뇌의 선조체 중 신경전달물질의 동시분석시 최적 조건)

  • Kang, Jong-Seong;Mun, Min-Seon;Shin, Hyung-Seon;Lee, Soon-Chul
    • Analytical Science and Technology
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    • v.8 no.2
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    • pp.215-220
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    • 1995
  • A simple, efficient and sensitive method was described for the simultaneous determination of catecholamine, indoleamine and related metabolites from the homogenates of the rat brain striatum by HPLC-ECD. The optimum mobile phase on a reverse phase $C_{18}$ column was 35mM sodium acetate buffer(included 10mM citric acid, 0.13mM $Na_4EDTA$, 0.58mM SOS, pH3-4):MeOH=85:15. The column temperature was $30^{\circ}C$. Dopamine(DA), 3, 4-dihydroxyphenyl acetic acid(DOPAC), homovanilic acid(HVA), 5-hydroxyindole acetic acid(5-HIAA), serotonin(5-HT) and noradrenaline(NA) could be separated and analysed to very small amount. The detection limits of this method were 2~10pg per injection for all components. The effects of age and sex of rat on the contents of the catecholamines and their metabolites in rat brain striatum were studied. The levels of DA and 5-HT contents of the 7 week old female rats were higher than those of the 7 week old male rats. As the age of rat increases, the contents of DOPAC increased significantly.

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Triglyceride Composition of Some Vegetable Oils 1. Triglyceride Composition of Sesame Oil (식물유의 Triglyceride 조성 1. 참기름의 Triglyceride 조성)

  • Park Yeung-Ho;WADA Shun;KOIZUMI Chiaki
    • Korean Journal of Fisheries and Aquatic Sciences
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    • v.14 no.1
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    • pp.1-6
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    • 1981
  • In this study, sesame oil was chosen as the experimental sample and analysed for its triglyceride composition by high-performance liquid chromatography(HPLC) in combination with gas liquid chromatography(GLC). The triglycerides were separated from sesame oil by liquid chromatographies on Bio-Beads SX-2 and on Sephadex LH-20, and fractionated into five groups on the basis of their partition numbers by reverse phase HPLC on a column packed with $\mu-Bondapak$ C18 using methanol-chloroform mix-ture as a solvent. Each of these collected fractions gave one to three peaks in the GLC chromatograms according to the acyl carbon number of the triglyceride, and fatty acid composition of the triglyceride was also analysed by GLC. From the results, it was found that the sesame oil consists with twenty one kinds of triglyceri-des, and the major triglycerides of sesame oil are those of $(2\;{\times}\;C18:1,\;C18:2\;;\;17.1\%),\;(C18:1,\;2{\times}C18:2\;;\;17.0\%),$ $(3\;{\times}\;C18:2\;;\;17.0\%),\;(3\;{\times}\;C18:1\;;\;10.9\%),$ $(3\;{\times}\;C18:2\;;\;9.6\%),\;(C16:0,\;C18:1,C18:2\;;\;7.9\%),$ $(C16:0,\;2\;{\times}\;C18:1\;;\;7.4\%),\;(C16:0,\;2\;{\times}\;C18:2\;;\;6.8\%),$ $(C18:0,\;C18:1,\;C18:2\;;\;3.1\%),\;(2\;{\times}\;C18:0,\;C18:2\;;\;1.5\%)$ $(C18:0,\;2\;{\times}\;C18:1\;;\;1.4\%),\;(C16:0,\;C18:0,\;C18:1\;;\;1.3\%),$ $(2\;{\times}\;C16:0,\;C18:1\;;\;1.2\%),\;and\;(C16:0,C18:0,\;C18:2\;;\;1.0\%)$.

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Variation of Bioactive Component Contents in Plant Parts of Paeonia lactiflora Pall. (작약 식물체 부위별 성분 함량 변이)

  • Choung, Myoung-Gun
    • Korean Journal of Medicinal Crop Science
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    • v.10 no.5
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    • pp.392-398
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    • 2002
  • Comparative analysis of paeoniflorin, albiflorin and phenolic compound contents as bioactive components of peony was performed by Reverse Phase-High Performance Liquid Chromatography (RPHPLC) using the four- year-old peony which were different plant parts and pretreatment, such as removing or unremoving the cork layer of peony root before drying. The contents of paeoniflorin, albiflorin, (+)-taxifolin $3-O-{\beta}-D-glucopyranoside$, (+)-catechin and (-)-epicatechin were the highest in rhizome part, but those of gallic acid and benzoic acid in the leaves were higher than other parts. The contents of albiflorin, gallic acid, benzoic acid and (-)-epicatechin in the cork layer were higher than in those of the core, but the contents of paeoniflorin, (+)-taxifolin $3-O-{\beta}-D-glucopyranoside$ and (+)-catechin in the core were higher than those in the cork layer. In general, the rhizome part of peony root has been used only propagation purpose, but this part contained high contents of bioactive component. Therefore, it is needed that medicinal application of rhizome part in peony root was firmly investigated. Also, In the use of peony root for medicinal purpose, the use of peony root with cork layer can be efficient way on the practical use of useful components and the reduction of labor for removing the cork layer.

Development of Analysis Method of Gardenia Yellow as Natural Colorants and Content Survey in Commercial Foods by HPLC (HPLC를 이용한 천연착색료인 치자황색소의 분석법 개발 및 시판 식품중 함유량조사)

  • Kim, Hee-Yun;Kim, So-Hee;Hong, Ki-Hyoung;Lee, Chul-Won;Kim, Kil-Saeng;Ha, Sang-Chul;Jo, Jae-Sun
    • Korean Journal of Food Science and Technology
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    • v.31 no.4
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    • pp.945-951
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    • 1999
  • A simple and practical method for the determination of gardenia yellow in foods was developed. In this method, analysis of gardenia yellow in food products has been carried out by the detection of crocetin and/or geniposide as indicator compounds. As a new analytical method for gardenia yellow, we adopted crocetin, which is produced from colored components of gardenia yellow by alkaline hydrolysis, as an indicator compound. The analysis of gardenia yellow was performed by reverse phase high performance liquid chromatography using a Capcell pak $C_{18}$ column at wave length 240 nm (geniposide) and 435 nm (crocetin). The recovery rates of geniposide and crocetin were found to be 93.4% and 87.8% for Dan Mu Ji, 90.2% and 85.9% for milk, 92.8% and 86.5% for snack, respectively. With this method, the range of crocetin and geniposide contents $({\mu}g/g)$ were as follows: $ND{\sim}1.7$ and $ND{\sim}14.1$ for Dan Mu Ji, $ND{\sim}0.2$ and $ND{\sim}13.6$ for milk, $ND{\sim}1.6$ and $ND{\sim}0.9$ for snack, respectively. The detection limits of crocetin and geniposide were 0.07 ${\mu}g/g$ and 0.05 ${\mu}g/g$, respectively.

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