• Title/Summary/Keyword: HPLC Chromatography

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Determination of enalapril in human blood by high-performance liquid chromatography mass spectrometer.

  • Chang, Dong-Jin;Shim, Chang-Koo;Chung, Suk-Jae
    • Proceedings of the PSK Conference
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    • 2002.10a
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    • pp.418.3-419
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    • 2002
  • Enalapril. a prodrug. is the ethyl ester of a long-acting angiotensin converting enzyme inhibitor. enalaprilat. Because enalapril does not contain any appreciable chromophore. detection of the drug in a complex matrix (e.g.. biological fluids) has been problematic with conventional detection systems in high-performance liquid chromatography (HPLC). As a result. determination of enalaprillevel in blood samples has been typically carried out using HPLC-MS/MS in the literature. (omitted)

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Isolation of Antimicrobial Substance from the Korean Traditional Leaf Mustard, Brassica juncea Coss.

  • Kang Seong-Koo
    • Plant Resources
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    • v.8 no.2
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    • pp.145-154
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    • 2005
  • The antimicrobial effect of each fraction after fractionation of an ethanol extract of leaf mustard was examined in terms of nucleic acid, chloroform, ethylacetate, and butanol. The ethylacetate fraction, which showed the strongest level of antimicrobial effect among the different ethanol extract fractions of leaf mustard, was isolated and purified using silica gel column chromatography and HPLC, respectively, to obtain a single antimicrobial substance called KLM-1. The antimicrobial effect of this substance was 10 times higher than that of the ethylacetate fraction. A further study is on the way to confirm the structure of the antimicrobial substance KLM-1 through LC/Mass and NMR.

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Detection of Undeclared Betamethasone Derivatives in Cosmetic Products Labeled to Contain Zinc Pyrithione as the Active Ingredient (아연피리치온을 유효성분으로 표기한 화장품류에서 미표기 성분인 베타메타손 유도체의 검출)

  • Lee, Jeong-Pyo;Park, Sung-Hwan;Yang, Seong-Jun;Kim, Sun-Mi;Son, Kyung-Hun;Yun, Mi-Ok;Choi, Sang-Sook
    • Journal of the Society of Cosmetic Scientists of Korea
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    • v.35 no.1
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    • pp.11-17
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    • 2009
  • Betamethasone propionate, an anti-inflammatory glucocorticosteroid, was detected in cosmetics with no indication on the label of this compound as an ingredient. The product was formulated as a topical spray or shampoo and labeled to contain zinc pyrithione as the active ingredient. A thin-layer chromatographic analysis was carried out on silica gel plates to provide a first indication about the presence of a compound with steroid structure and reactivity; then high-performance liquid chromatography (HPLC) separation allowed the identification of the corticosteroid agent and its quantification. To identify the corticosteroid agent from these commercial samples we collected the fractions suspected to have ketol steroids by prep HPLC and identified the compound as betamethasone propionate by NMR and MS spectrometry. Then we synthesized the standard for the betamethasone 17-propionate and 21-propionate and quantitate the corticosteroids from the sample by HPLC with that standards. By this method we identified the corticosteroid compounds from some commercial cosmetics such as zinc pyrithione sprays. The finding of betamethasone propionate in the products was shown by comparison to an authenticated standard of betamethasone propionate by retention time on reverse-phase HPLC. Two of the tested products contained betamethasone propionate at the levels of 0.005 ${\sim}$ 0.02% and the others were free of betamethasone propionate.

Isolation of 3,4-Dihydroxybenzoic Acid with Antimicrobial Activity from Bark of Aralia elata (두릅수피에서 항미생물활성을 갖는 3,4- dihydroxybenzoic acid의 분리)

  • Ma, Seung-Jin;Ko, Byoung-Seub;Park, Keun-Hyung
    • Korean Journal of Food Science and Technology
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    • v.27 no.5
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    • pp.807-812
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    • 1995
  • The methanol extracts of Aralia elata bark showed antimicrobial activities against bacteria, yeast, fungi. The solvent fractionated acidic fraction that showed the activity was then successively purified with silica gel adsorption column chromatography, Sephadex LH-20 column chromatography, silica gel partition column chromatography, HPLC and TLC. The isolated major active substance was identified as 3,4-dihydroxybenzoic acid by MS, GC-MS, IR, $^{1}H-NMR\;and\;^{13}C-NMR$. The content of 3,4-dihydroxybenzoic acid was 0.869㎎/g in dried bark of Aralia elata.

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Antimicrobial Substances in Leek (Allium tuberosum) (부추의 항미생물 활성물질)

  • Kim, Seon-Jae;Park, Keun-Hyung
    • Korean Journal of Food Science and Technology
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    • v.28 no.3
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    • pp.604-608
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    • 1996
  • The antimicrobial activty of leek (Allium tuberosum) was investigated against 17 strains of microorganisms. Methanol extracts of leek showed the growth inhibition effects on the wide range of microorganisms including gram positive bacteria, gram negative bacteria and yeasts. The extracts were analysed by using solvent fractionation, silica gel adsorption chromatography, Sephadex LH-20 column chromatography, TLC, silica gel partition chromatography and HPLC techniques. Six components whose molecular weights range from 200 to 400 were confirmed to have the antimicrobial activity.

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Antioxidative Activity of Hot Water and Ethanol Extracts of Lespedeza cuneata Seeds (야관문(Lespedeza cuneata) 종자의 열수 및 Ethanol 추출물의 항산화 효과)

  • Kim, Seon-Jae;Kim, Du-Woon
    • Food Science and Preservation
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    • v.14 no.3
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    • pp.332-335
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    • 2007
  • Hot water and 95%(v/v) ethanol extracts were prepared from dried Lespedeza cuneata seeds and antioxidant compounds were isolated by solvent fractionation, silica gel adslorption chromatography, Sephadex LH-20 column chromatography, and preparative HPLC. Antioxidant activity was measured using DPPH radical scavenging activity. The 80%(v/v)ethanol and ethylacetate fraction of Lespedeza cuneata seed extracts had stronger antioxidant effects than did the n-hexane fraction. The active antioxidant compounds obtained from hot water and 95%(v/v) ethanol extracts may be identical, based on analysis by Sephadex LH-20 column chromatography and preparative HPLC.

Avantor® ACE® Wide Pore HPLC Columns for the Separation and Purification of Proteins in Biopharmaceuticals (바이오의약품의 단백질 분리 및 정제를 위한 Avantor® ACE® 와이드 포어 HPLC 컬럼 가이드)

  • Matt James;Mark Fever;Tony Edge
    • FOCUS: LIFE SCIENCE
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    • no.1
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    • pp.3.1-3.7
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    • 2024
  • The article discusses the critical role of chromatography in the analysis and purification of proteins in biopharmaceuticals, emphasizing the importance of comprehensive characterization for ensuring their safety and efficacy. It highlights the use of Avantor® ACE® HPLC columns for the separation and purification of proteins, focusing on the analysis of intact proteins using reversed-phase liquid chromatography (RPLC) with fully porous particles. This article also details the application of different mobile phase additives, such as TFA and formic acid, and emphasizes the advantages of using type B ultra-pure silica-based columns for efficiency and peak shape in biomolecule analysis. Additionally, it addresses the challenges of analyzing intact proteins due to slow molecular diffusion and introduces the concept of solid-core (or superficially porous) particles, emphasizing their benefits over traditional porous particles for the analysis of therapeutic proteins. Furthermore, it discusses the development of Avantor® ACE® UltraCore BIO columns, specifically designed for the high-efficiency separation of large biomolecules, such as proteins, and demonstrates their effectiveness in achieving high-resolution separations, even for higher molecular weight proteins like monoclonal antibodies (mAbs). In addition, it underscores the complexity of analyzing and characterizing intact protein biopharmaceuticals, requiring a range of analytical techniques and the use of wide-pore stationary phases, operated at elevated temperatures and with relatively shallow gradients. It highlights the comprehensive range of options offered by Avantor® ACE® wide pore columns, including both fully porous and solid-core particles, bonded with a variety of complementary stationary phase chemistries to optimize selectivity during method development. The use of ultrapure and highly inert base silica is emphasized for enabling the use of lower concentrations of mobile phase modifiers without compromising analyte peak shape, particularly beneficial for LC-MS applications. Then the article concludes by emphasizing the significance of reversed-phase liquid chromatography and its compatibility with mass spectrometry as a valuable tool for the separation and analysis of intact proteins and their closely related variants in biopharmaceuticals.

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Isolation and Purification of Berberine in Cortex Phellodendri by Centrifugal Partition Chromatography (Centrifugal Partition Chromatography에 의한 황백으로부터 Berberine의 분리 및 정제)

  • Kim, Jung-Bae;Bang, Byung-Ho
    • The Korean Journal of Food And Nutrition
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    • v.27 no.3
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    • pp.532-537
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    • 2014
  • Cortex Phellodendri (CP) is derived from the dried bark of Phellodendron amurense. It has been widely used as a drug in traditional Korea medicine for treating diarrhea, jaundice, swelling pains in the knees and feet, urinary tract infections, and infections of the body surface. Many analytical methods have been used to study oriental herbal medicines, such as thin-layer chromatography, column liquid chromatography, and high performance liquid chromatography (HPLC). In this study, preparative centrifugal partition chromatography (CPC) was successfully carried out in order to separate pure compounds from a CP methanol extract. The optimum two-phase CPC solvent system was composed of n-butanol: acetic acid: water (4:1:5 v/v/v). The flow rate of the mobile phase was 3 mL/min in ascending mode with rotation at 1,000 rpm. The CPC-separated fraction and purification procedures were carried out by preparatory HPLC. The $^1H$ NMR spectrum revealed that the resonances at ${\delta}$ 4.10 and 4.20 ppm corresponded to three protons ($-OCH_3$), whereas those at ${\delta}$ 6.10 ppm corresponded to two protons ($-OCH_2O-$). Further, two aromatic protons (H-11 and H-12) conveys a doublet-doublet pattern. The H-11 doublet and H-12 doublet appear at ${\delta}$ 7.98 and 8.11, respectively. The $^{13}C$ NMR. spectrum showed a tetrasubstituted with a methylenedioxy group at C2 and C3, and two methoxy groups at C9 and C10. The chemical structure of the berberine was identified by $^1H$, $^{13}C$-nuclear magnetic resonance and electrospray ionization-mass spectroscopy spectral data analysis.

Chemiluminescence immunochromatographic analysis for the quantitative determination of algal toxins

  • Pyo, Dongjin;Kim, Taehoon
    • ALGAE
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    • v.28 no.3
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    • pp.289-296
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    • 2013
  • For the quantitative detection of algal toxin, microcystin, a chemiluminescence immunochromatographic assay method was developed. The developed system consists of four parts, chemiluminescence assay strip (nitrocellulose membrane), horse radish peroxidase labeled microcystin monoclonal antibodies, chemiluminescence substrate (luminol and hydrogen peroxide), and luminometer. The performance of the chemiluminescence immunochromatographic assay system was compared with high performance liquid chromatography (HPLC) detection. The detection limit of chemiluminescence immunochromatographic assay system is several orders of magnitude lower than with HPLC. The chemiluminescence immunochromatography and HPLC results correlated very well with the correlation coefficient ($r^2$) of 0.979.

HPLC Resolution of Enantiomers Using Polysaccharide Derivatives as Chiral Stationay Phases

  • Okamoto, Yoshio
    • Proceedings of the PSK Conference
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    • 2002.10a
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    • pp.163-164
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    • 2002
  • In the past two decades, separations of enantiomers (optical isomers) by high-performance liquid chromatography (HPLC) have remarkably advanced [1]. Among many commercially available chiral stationary phases (CSPs) for HPLC, polysaccharide-based CSPs are the most popular ones, which can cover the resolution of a wide range of the chiral compounds [2, 3, 4]. Here, I will explain mainly the HPLC separation of enantiomers using these CSPs. (omitted)

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