• 제목/요약/키워드: HPLC(high performance liquid chromatography)

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젤 여과, 이온 크로마토그래피와 HPLC에 의한 효모 엑기스내의 비타민의 분석연구 (Characterization of Vitamins in Yeast Extract using Gel Filtration, Ion Exchange Chromatography and HPLC)

  • 최인호;홍억기;강환구;김인호
    • KSBB Journal
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    • 제15권1호
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    • pp.76-79
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    • 2000
  • Complex, ill-defined mixtures of natural origin are often used as nutrients in the production of biological products through microbial fermentation. Product yields are affected by variation in these natural products. Yeast extract is a typical example of these natural products. Since it is a mixture of amino acids, peptides and nucleic acids, its composition is not well characterized. In this study, we investigated the properties of thiamine hydrochloride, riboflavin and pyridoxine hydrochlride in yeast extract by using a gel filtration chromatography, ion exchange chromatography and high performance liquid chromatography. Yeast extract solution was fractionated by gel filtration chromatography and ion exchange chromatography, and then, each fraction was analyzed by using a high performance liquid chromatography.

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Simultaneous Determination of Cysteamine and Cystamine in Cosmetics by Ion-Pairing Reversed-Phase High-Performance Liquid Chromatography

  • Kim, Yejin;Na, Dong Hee
    • Toxicological Research
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    • 제35권2호
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    • pp.161-165
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    • 2019
  • Cysteamine has been used in cosmetics as an antioxidant, a hair straightening agent, and a hair waving agent. However, recent studies indicate that cysteamine can act as an allergen to hairdressers. The objective of this study was to develop and validate a simple and effective reversed-phase high-performance liquid chromatography (RP-HPLC) method for the measurement of cysteamine and its dimer, cystamine. Sodium 1-heptanesulfonate (NaHpSO) was used as an ion-pairing agent to improve chromatographic performance. Separation was performed on a Gemini C18 column ($250mm{\times}4.6mm$, $5{\mu}m$ particle size) using a mobile phase composed of 85:15 (v/v) 4 mM NaHpSO in 0.1% phosphoric acid:acetonitrile. UV absorbance was monitored at 215 nm. The RP-HPLC method developed in this study was validated for specificity, linearity, limit of detection, limit of quantitation, precision, accuracy, and recovery. Cysteamine and cystamine were chromatographically resolved from other reducing agents such as thioglycolic acid and cysteine. Extraction using water and chloroform resulted in the recovery for cysteamine and cystamine ranging from 100.2-102.7% and 90.6-98.7%, respectively. This validated RP-HPLC method would be useful for quality control and monitoring of cysteamine and cystamine in cosmetics.

HPLC-MSD 를 이용한 식육 중 합성항균제의 동시분석 (Simultaneous detection for synthetic antimicrobials in muscle by high performance liquid chromatography-mass selective detector (HPLC-MSD))

  • 홍인석;최윤화;권택부;이정학
    • 한국동물위생학회지
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    • 제29권3호
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    • pp.317-330
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    • 2006
  • This study was conducted to develop the analytical method about simultaneous determination for synthetic antimicrobials in muscle by high performance liquid chromatography - mass selective detector (HPLC- MSD). Solid phase extraction (SPE), matrix solid phase dispersion (MSPD) and liquid-liquid extraction (LLE) have been adapted as pretreatment procedures for HPLC- MSD. Among various solvent tested, methanol was chosen for extraction of synthetic antimicrobials in muscles. For the optimized response, the values of various MS parameters including fragment voltage, drying gas flow, nebulizer pressure, drying gas temperature were verified. The average recovery rates using MSPD and SPE for muscles of bovine and pork were 78.9-127.1% and 78.3-121.7%, respectively. This method was verified the satisfactory performance for fourteen synthetic antimicrobials excepting carbadox in muscle of pork as detection limit of $0.05{\mu}g/g$ on API/ES SIM mode.

고속역류크로마토그래피 기술을 이용한 생리활성 물질의 분리 및 정제 (Isolation and Purification of Bioactive Materials Using High-Performance Counter-Current Chromatography (HPCCC))

  • 정동수;신현재
    • KSBB Journal
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    • 제25권3호
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    • pp.205-214
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    • 2010
  • 역류크로마토그래피 (counter-current chromatography, CCC)는 일련의 분배과정을 한 개의 튜브 내에서 연속적으로 일어나도록 고안된 시스템으로서 컬럼으로는 polytetrafluoroethylene(PTFE) 튜브가 다층으로 감겨있는 원통형의 홀더 3개가 서로 기어를 통해 물려있으며, 홀더가 회전과 공전을 통해 튜브의 꼬임을 방지하는 rotary seal-free flow centrifuge 시스템으로 되어있다. 역상 HPLC (reverse phase HPLC)에서는 고정상이 실리카에 결합된 유기물단 (organic moiety)이 수용성 이동상 물질에 의해서 용매화 (solvated)되는 반면 CCC는 실리카 대신에 강한 중력장에 의해 분리되는 자유로운 용매가 고정상이 되며 이 고정상의 부피비율은 20-30%에 이른다. 즉 고체담체에 결합된 유기관능기 대신에 물과 섞이지 않는 hexane 같은 유기용제가 고정상으로 사용되는 것이다. 고속역류크로마토그래피 (high-performance countercurrent chromatography, HPCCC)는 CCC의 기능을 향상시킨 분리시스템으로서 높은 중력장하에서 높은 이동상 속도와 높은 분리효율과 짧은 분리시간을 특징으로 하고 있다. 특히 mg 단위에서 kg 단위로의 스케일업이 선형적으로 가능하다는 큰 장점을 지니고 있다. 이 총설에서는 현재까지 개발된 CCC의 일반적인 이론을 간략히 정리하고 최신 HPCCC 장비의 적용 예를 살펴보고 그 응용분야로서 생리활성물질의 분리 및 정제와 관련된 연구동향을 정리하였다.

HPLC를 이용한 식육내 타이로신의 잔류분석법 (Determination of tylosin in edible meats by high-performance liquid chromatography)

  • 김곤섭;신선혜;김종수;라도경
    • 대한수의학회지
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    • 제41권1호
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    • pp.13-19
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    • 2001
  • A simple and rapid analytical method for the determination of tylosin in chicken, pork and muscle was established by High-Performance Liquid Chromatography(HPLC). Chicken, pork and beef muscle(5 g) were fortified by adding the $0.2{\mu}g/ml$ of standard tylosin and the drug was extracted from meats with 70% acetonitrile(ACN) and followed by liquid-liquid partition for clean-up procedure. Then $20{\mu}l$ portion of ACN elution was directly analyzed by HPLC with spectra 100 variable wavelength detector, and unfortified blank control were treated similarly. The average recovery rate of tylosin added to chicken, pork and beef muscle were $83{\pm}2.3$, $96{\pm}3.3$ and $92{\pm}1.6$(%) at the level 0.2 ppm, respectively. No tylosin residues in marketing meats. These results suggested that HPLC methodology could be acceptable for the extraction, determination and screening of tylosin residues in edible meats.

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High-Performance Liquid Chromatographic Determination of Cyclosulfamuron Residues in Soil, Water, Rice Grain and Straw

  • Lee, Young-Deuk;Kwon, Chan-Hyeok
    • 한국환경농학회지
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    • 제23권4호
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    • pp.251-257
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    • 2004
  • Analytical methods were developed to determine cyclosulfamuron residues in soil, water, rice grain and straw using high-performance liquid chromatography (HPLC) with ultraviolet absorption detection. In these methods, cyclosulfamuron was extracted with aqueous $Na_2HPO_4$/acetone and acetone/methanol mixture from soil and rice samples respectively. Liquid-liquid partition coupled with ion-associated technique, Florisil column chromatography, and solid-phase extraction (SPE) were used to separate cyclosulfamuron from interfering co-extractives prior to HPLC analysis. For water sample, the residue was enriched in $C_{18}$-SPE cartridge, cleaned up in situ, and directly subjected to HPLC. Reverse-phase HPLC under ion-suppression was successfully applied to determine cyclo-sulfamuron in sample extracts with the detection at its ${\lambda}_{max}$ (254 nm). Recoveries from fortified samples averaged $87.8{\pm}7.1%$ (n=12), $97.3{\pm}7.2%$ (n=12), $90.8{\pm}6.6%$ (n=6), and $78.5{\pm}6.7%$ (n=6) for soil, water, rice grain and straw, respectively. Detection limits of the methods were 0.004 mg/kg, 0.001 mg/L, 0.01 mg/kg and 0.02 mg/kg for soil, water, rice grain and straw samples, respectively.

Determination of isovitexin from Lespedeza cuneata using a validated HPLC-UV method

  • Lee, Ju Sung;Paje, Leo Adrianne;Yoo, Sang-Woo;Lee, Seong;Ku, Ja-Jung;Lee, Sanghyun
    • Journal of Applied Biological Chemistry
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    • 제64권1호
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    • pp.63-67
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    • 2021
  • Isovitexin, a marker compound with various pharmacological activities, in Lespedeza cuneata, was analyzed using high performance liquid chromatography coupled with UV (HPLC/UV). There are no previous reports on using L. cuneata as the source material for the quantification of isovitexin. In this study, we developed an optimized method using HPLC-UV analysis, which was validated using various parameters. Our method demonstrated high specificity, and good separation of the chromatographic peak was achieved. Parameters such as linearity (r2≥0.9997), precision, and accuracy indicated that our proposed analytical method had good reliability and sensitivity. These results demonstrate the utility and convenience of our method for rapidly quantifying isovitexin in L. cuneata extracts.

High-Performance Liquid Chromatographic Determination of Tricyclazole Residues in Rice Grain, Rice Straw, and Soil

  • Lee, Young-Deuk;Lee, Jung-Hun
    • Applied Biological Chemistry
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    • 제41권8호
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    • pp.595-599
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    • 1998
  • An analytical method was developed to determine tricyclazole residues in rice grain, straw, and soil using high-performance liquid chromatography (HPLC) with ultraviolet absorption detection. Tricyclazole was extracted with methanol from moist rice grain, straw, and soil samples. n-Hexane washing was employed to remove nonpolar co-extractives during liquid-liquid partition. Tricyclazole was then extracted with dichloromethane from alkaline aqueous phase, while acidic interferences remained in the phase. Dichloromethane extract was further purified by silica gel column chromatography prior to HPLC determination. Reverse-phase HPLC using an octadecylsilyl column was successfully applied to separate and quantitate the tricyclazole residue in sample extracts monitored at ${\lambda}_{max}$ 225nm. Recoveries from fortified samples averaged $95.5{\pm}3.0%\;(n=6),\;87.5{\pm}20.%\;(n=6),\;and\;84.3{\pm}2.8%$ (n=12) for rice grain, straw, and soil, respectively. Detection limit of the method was 0.02 mg/kg for rice grain and soil samples while 0.05 mg/kg for rice straw samples. The proposed method was reproducible and sensitive enough to evaluate the safety of tricyclazole residues in rice grain, straw, and soil.

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Quality Evaluation of Modified Bo-Yang-Hwan-O-Tang by Capillary Electrophoresis and High-performance Liquid Chromatography

  • Chen, Jianbo;Wu, Enqi;Zhu, Hongmei;Lee, Kwan-Jun;Chu, Van Men;Cho, Cheong-Weon;Kim, Young-Ho;Park, Yong-Ki;Lee, Won-Jae;Kang, Jong-Seong
    • Bulletin of the Korean Chemical Society
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    • 제32권8호
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    • pp.2666-2670
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    • 2011
  • High-performance liquid chromatography (HPLC) and capillary electrophoresis (CE) were used to identify five active components in the modified herbal decoction Bo-Yang-Hwan-O-Tang (mBHT), i.e., amygdalin, decursin, paeoniflorin, salvianolic acid B, and calycosin-7-O-${\beta}$-D-glycoside. These components were identified by comparing their retention times and mass spectra with those of reference compounds. The conditions of both analytical methods were optimized and validated. Sufficient separation of target analytes was achieved using a buffer consisting of 40 mM sodium borate and 60 mM sodium dodecylsulfate (SDS) containing 10% methanol (pH 9.5) at 250 nm for CE analysis and gradient elution with a water-methanol mobile phase and ultraviolet (UV) photodiode array detector (DAD) at 250 nm for HPLC analysis. The mBHT components were determined within 65 min by HPLC and 16 min by CE. All calibration curves showed high linearity (R > 0.999) within the ranges tested. Intra-day and inter-day precision were less than 1.6% and 1.8% for HPLC and 2.5% and 4.8% for CE, respectively. The accuracy of the methods ranged from 98.8% to 102.3% for HPLC and from 95.9% to 108.2% for CE.