• Title/Summary/Keyword: Reversed-phase HPLC

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HPLC Determination of Carboxyl Grop usinf 2-Bromoacetyltriphenylene as Pre-labeling Reagent (III) - Separative determination of glycyrrhetinic acid contained in licorice power (2-Bromoacetyltriphenylene 유도체화제를 이용한 카르복실기 함유성분의 분석법 (III) - 감초중 glycyrrhetinic acid의 HPLC에 의한 분리정량)

  • 정해수;예덕천;김박광;박만기;이왕규
    • YAKHAK HOEJI
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    • v.31 no.5
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    • pp.315-321
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    • 1987
  • A high performance liquid chromatographic method was developed for the determination of glycyrrhetinic acid contained in licorice powder. Glycyrrhetinic acid which is hydrolysate of glycyrrhizin extracted from licorice powder, was determined with good result by HPLC using 2-bromoacetyltriphenylene labeling reagent. The glycyrrhetinic acids were labeled with 2-bromoacetyltriphenylene in acetonitrile using 18-crown-6-ether and KOH as a catalyst. Derivatized glycyrrhetinic acids were separated from the extracted licorice powder on a reversed-phase column (chemopak $C_{18}$) using 100% acetonitrile as a mobile phase and monitored by an UV-detector at 268nm. Linearity of calibration curve was obtained between 5 ng and 20 ng, and the lower limit of detection was 2 ng. The recovery of glycyrrhetinic acid to licorice powder was about 99.3%. This method was sensitive, reliable and useful for, determination of glycyrrhetinic acid.

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Determination and Ethylene Oxide(EO) Separation of Polysorbates by RP-HPLC (역상 HPLC에 의한 polysorbates의 산화에틸렌(EO) 분리 및 정량분석)

  • Lee, Yong-Hwa
    • Journal of the Korean Applied Science and Technology
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    • v.29 no.4
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    • pp.585-593
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    • 2012
  • The EO separation and quantitative determination of polysorbate 20, polysorbate 40, polysorbate 60 and polysorbate 80 was carried out by reversed phase HPLC. The water/acetonitrile was used for the mobile phase of gradient conditions. An YMC Pack Ph ($250mm{\times}4.6mm$ i.d., $5{\mu}m$) and Phenomenex C4 ($250mm{\times}4.6mm$ i.d., $5{\mu}m$) and the selected ELSD detector was applied. The analysis results of HPLC showed good linearity with correlation coefficient of $r^2$=0.997 in the rage of $180.2{\sim}980.5{\mu}g/mL$ and detection limit.

On-line Trace Enrichment for the Determination of Insulin in Biological Samples Using Reversed-Phase High Performance Liquid Chromatography with Column Switching

  • Lee, Jung-Sook;Lee, Heeyong;Lee, Hye-Suk;Lee, Kang-Choon
    • Archives of Pharmacal Research
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    • v.17 no.5
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    • pp.360-363
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    • 1994
  • Column--swtiching technique with a reversed-phase high performance liquid chromatographic method has been developed for the routine analysis of radioiodinated insulin and its degadation products in biological fluids. The diluted biological samples were loaded onto a precolumn packed with LiChrosorb RP-8 $(25-40{\;}{\mu}m)$ using 0.1% trifuoroacetic acid (TFA) in water as a washing solvent. After valve switching, the concentrated insulins were eluted in the back-flush mode and separated by a W-Porex $C_{18}$ column with a gradient of 0.1% TFA in water and 0.1% TFA in acetonitrile as the mobile phase. The method showed good precision, accuracy and speed with the detection limit of 20 pg/ml. Total analysis time per sample was about 40 min and the coefficients of variation were less than 8, 2%.

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Analysis of Flavonoid Components from Machilus thunbergii Leaves (후박나무 잎 Flavonoid 성분의 분석)

  • 김성환
    • Journal of Plant Biology
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    • v.36 no.3
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    • pp.297-300
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    • 1993
  • Six flavonoid compounds from the ethyl acetate fraction of Machilus thunbergii leaves were identified by HPLC. Separation by reversed phase chromatography on u-Bondapak C18 column was achieved by isocratic elution. The contents of the major flavonoids, guaijaverin and quercitrin were about 0.48% (w/w) and 0.98% (w/w) for the methanol extract, respectively.

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Simultaneous Determination of Tin, Nickel, Lead, Cadmium and Mercury in Cigarette Material by Solid Phase Extraction and HPLC

  • Hu, Qun;Yang, Guangyu;Ma, Jing;Liu, Jikai
    • Bulletin of the Korean Chemical Society
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    • v.24 no.10
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    • pp.1433-1436
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    • 2003
  • A new method for the simultaneous determination of heavy metal ions in cigarette material by microwave digestion and reversed-phase high-performance liquid chromatography (RP-HPLC) has been developed. The cigarette material was digested by microwave digestion. Lead, cadmium, mercury, nickel and tin ions in the digested samples were pre-column derivatized with tetra-(2-chlorophenyl)-porphyrin ($T_2$-CPP) to form color chelates, which were then enriched by solid phase extraction with a $C_{18}$ cartridge. The chelates were separated on a Waters Xterra$^{TM}RP_{18}$ column by gradient elution with methanol (containing 0.05 mol/L pyrrolidine-aceticacid buffer salt, pH = 10.0) and acetone (containin0.05 mol/L pyrrolidine-acetic acid buffer salt, pH = 10.0)as mobile phase at a flow rate of 0.5mL/min and analyzed with a photodiode array detector from 350-600 nm. The detection limits of lead, cadmium, mercury, nickel and tin were 4,3,3,8 and 5 ng/L, respectively, in the original samples. This method was afforded good results.

Reverse-Phase HPLC Method for Identification of Diastereomeric Constituents from Sasa borealis (Sasa borealis의 Diastereomeric 성분들의 역상 고속액체크로마토그래프 분석방법)

  • Jeong Yeon Hee;Lee Jun;Kwon Youngjoo;Seo Eun-Hyoung
    • YAKHAK HOEJI
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    • v.50 no.1
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    • pp.21-25
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    • 2006
  • Reiterated normal-phase column chromatography lead to the isolation and purification of six known compounds but for the first time from the whole plant of Sasa borealis (Hack.) Makino (Gramineae): tricin 4'-O-(erythro-${\beta}$-guaia-cylglyceryl) ether (1), tricin 4'-O-(threo-${\beta}$-guaiacylglyceryl) ether (2), tricin 4'-O-[erythro-${\beta}$-guaiacyl-(9'-O-acetyl)-glyceryl] ether (3), tricin 4'-O-[threo-${\beta}$-guaiacyl-(9'-O-acetyl)-glyceryl] ether (4), (-)-pinoresinol (5), and vanillin (6). The structures of the compounds (1-6) were established based on interpretation of high resolution NMR (COSY, HSQC, HMBC, and NOESY) spectral data. In particular, compounds 1 and 3 were diastereomers of compounds 2 and 4, respectively. These two sets of diastereomers were able to be simultaneously identified and quantified by a gradient reversed-phase HPLC method with UV photodiode array, This sensitive HPLC method is noteworthy as a simultaneous separation and identification method to test the extract of the family Gramineae which contains these compounds.

Retention Behavior of Organic Compounds on Reversed-Phase Column expected by Van der Waals Volume (Van der Waals Volume을 이용한 역상 컬럼에서의 유기화합물들의 용출거동)

  • Park, Wun-Kyu;Lee, Yong-Moon;Moon, Dong-Cheul;Kang, Jong-Seong
    • Analytical Science and Technology
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    • v.6 no.4
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    • pp.383-390
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    • 1993
  • Retention behaviors of aliphatic and aromatic compounds were investigated with Van der Waals volume which represented the molecular size. The organic solvents, methanol, acetonitrile and tetrahydrofuran, were mixed with water at various ratio, respectively. The selectivity of organic solvents were tested by change of column temperature. The capacity factor was increased linearly according to the enlargement of molecular size. Therefore, Van der Waals volume was useful to predict the elution of organic molecules in reversed-phase column. The order of elution capacity of solvents was methanol

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Determination of Rosmarinic Acid and Caffeic Acid from Perilla frutescens var. japonica and var. acuta by Reversed-Phase HPLC (RP-HPLC를 이용한 백소엽(白蘇葉)과 자소엽(紫蘇葉)의 카페익산과 로즈마린산 분석)

  • Kim, Byung-Youn;Jeong, Ji-Seon;Kwon, Ha-Jeong;Lee, Je-Hyun;Hong, Seon-Pyo
    • The Korea Journal of Herbology
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    • v.23 no.3
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    • pp.67-72
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    • 2008
  • Objectives : This study presents a high performance liquid chromatography methods for the quantitative and qualitative analysis of rosmarinic acid (RA) and caffeic acid (CA) in Perilla frutescens var. japonica and var. acuta. Methods : Chromatographic separation was performed using a mixture of methanol, water and formic acid (35 : 64.2 : 0.8) with a reversed-phase column (Gemini C18, 4.6 ${\times}$ 150 mm, 3 ${\mu}m$). The analyses were detected at UV (280 nm). Results : The samples were extracted with 50% EtOH under reflux for 1 h, and simultaneous determination for RA and CA in hyang-so-san and haeng-so-san was possible without interference peaks Conclusions : According the results, we developed a determination method for RA and CA in Perillae Folium. Owing to Perilla frutescens var. japonica and var. acuta did not show significant difference in contents of RA and CA, both Perilla frutescens could be available as herbal medicine.

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Quantitative analysis of selenium species in sea food using solid phase extraction and HPLC-ICP/MS (해산물 시료에서 solid-phase extraction 및 HPLC-ICP/MS를 이용한 셀레늄 화학종의 정량분석)

  • Kim, Eunju;Joo, Minkyu;Kwon, Hyosik;Pak, Yongnam
    • Analytical Science and Technology
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    • v.26 no.5
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    • pp.307-314
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    • 2013
  • Selenium exists in various forms of chemical species. The activity and bioavailability is strongly dependent on its chemical form and concentration. Consequently the information on each selenium species and its concentration must be exactly determined for the food we take in. In this study, selenium species in seafood were separated and quantified by RP (reversed phase) HPLC (high performance liquid chromatography) coupled with ICP-MS (inductively coupled plasma mass spectrometry) using post-column isotope dilution. $^{79}Br$, which interferes on $^{80}Se$, has mostly been removed by solid phase extraction and then mathematical correction has been applied for the more accurate correction. The experimental result for CRM (certified reference material) DOLT-4 agreed well with the certified value but each selenium species could not be compared. SeCys (selenocysteine) and SeMet (selenomethionine) were the major species detected in seafood such as belt fish, spanish mackerel, and squid that have been serving as Korean diet. The concentrations found in Korean sea food for SeCys and SeMet were in the range of 0-661.6 mg/kg and 137.3-462.7 mg/kg, respectively.