• 제목/요약/키워드: Fluorescence method

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Drug-Biomacromolecule Interaction XII: Comparative binding study of sulfaethidole to bovine serum albumin by equilibrium dialysis, fluorescence probe technique, uv difference spectrophotometry and circular dichroism

  • Kim, Chong-Kook;Chun, Yang-Sook;Lah, Woon-Lyong
    • Archives of Pharmacal Research
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    • v.12 no.3
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    • pp.160-165
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    • 1989
  • Binding of sulfaethidole to bovine serum albumin was studied by equilibrium dialysis, fluorescence probe technique, uv difference spectrophotometry and circular dichroism. Equilibrium dialysis method enabled us to estimate the total number of drug binding sites of albumin molecule. For sulfaethidole, albumin had 6 primary and 40 secondary binding sites. The primary and secondary binding constants were 0.9 * 10/sup 5/ M/sup -1/ and 0.2 * 10/sup 6/ M/sup -1/, respectivitely. 1-Anilino-8-naphthalenesulfonate (ANS) and 2-(4-hydroxylbenzeneazo)- benzoic acid (HBAB) were used as the fluorescence probe and the uv spectrophotometric probe, respectively. In fluorescence probe technique, results indicated that the number of higher affinity drug binding site of albumin was 1 and the number of lower affinity drug binding sites of albumin was 3, and the primary and secondary drug binding constants for bovine serum albumin were 2.15 * 10/sup 5/M/sup -1/ and 1.04 * 10/sup 5/ M/sup -1/, respectively. In uv difference spectrophotometry, binding sites were 3 and binding constant was 1.88 * 10/sup 5/M/sup -1/. The above spectrophotometry, binding sites were 3 and binding constant was 1.88 * 10/sup 5/M/sup -1/. The above results suggest that several different methods should be used in ompensation for insufficient information about drug binding to albumin molecule given by only one method.

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Selective Analysis of Heavy Metal Ions Using Protein-based Biosensor (단백질 바이오센서를 이용한 중금속 이온의 선택적 측정)

  • 김균영;김지현;유영제
    • KSBB Journal
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    • v.16 no.6
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    • pp.609-613
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    • 2001
  • New protein-based biosensors using fluorescence for the detection heavy metal ions were developed. The detection range of heavy metal ions was between 10$\^$-3/ mM - 1 mM using casein and albumin as a transducer of biosensor, respectively. Casein showed better results for detecting heavy metal ions than albumin. Simple assay method was developed for the selective analysis of the two heavy metal ions by the fluorescence at wavelength of excitation and emission. This method was successfully applied to determining the concentrations Of Co$\^$2+/ and Fe$\^$3+/.

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Fluorimetric Determination of Phosphate in Sea Water by Flow Injection Analysis

  • Motomizu, Shoji;Oshima, Mitsuko;Katsumura, Naoya
    • Analytical Science and Technology
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    • v.8 no.4
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    • pp.843-848
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    • 1995
  • A sensitive method for the determination of trace amounts of phosphate by fluorescence-quenching detection / FIA is proposed. The fluorescence of Rhodamine B(RB) was quenched with the formation of the ion associate of molybdophosphate with RB;${\lambda}_{ex}$ and ${\lambda}_{em}$ were 560nm and 580nm, respectively. A calibration graph was linear over the ranges from $10^{-8}$ to $3{\times}10^{-6}M$ of phosphate (~0.3~93ppb of phosphorus). The relative standard deviation was 1.2% with $8{\times}10^{-7}M$ phosphate solution and sampling rate was 15 samples / h. The proposed method was applied to the determination of phosphate in sea and river water samples.

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Determination of Glyphosate in Whole Blood by HPLC-fluorescence Detection (HPLC 형광검출법에 의한 Glyphosate의 혈중농도 측정)

  • 이상기;김기욱;양자열;인상환;이수연
    • YAKHAK HOEJI
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    • v.45 no.4
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    • pp.347-351
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    • 2001
  • A rapid and sensitive method for the determination of glyphosate, a phosphated amino acid herbicide, in whole blood is presented. After removal of protein, the whale blood was purified by using the anion exchange resin (Dowex 1), and derivatized with 9-fluorenylmethyl chloroformate (FMCL). Derivatized glyphosate from blood sample was injected onto a Whatman partisil 10SAX column and separated with 0.1M phosphate buffer (pH 2.5) and acetonitrile (ratio=3:1). The high performance liquid chromatography-fluorescence detection gave the detection limit of 86pg and linearity of 0.9999 in the range of 0.25 $\mu$g/ml and 25 $\mu$g/ml. The recoveries of glyphosate added to the blood samples were ranged from 75.3% to 100.4% compared to the samples prepared in water. The derivatized glyphosate was stable at various acidity and temperature. This method has been successfully applied to the blood samples of lethal intoxication with the herbicide glyphosate.

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Characterization of High Pressure-High Temperature Treated Gem Diamonds (고압고온 처리된 보석용 다이아몬드의 감별 연구)

  • Song, Oh-Sung
    • Journal of Surface Science and Engineering
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    • v.39 no.5
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    • pp.229-234
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    • 2006
  • Diamonds have been widely employed as polishing media for precise machining and noble substrates for microelectronics. The recent development of the split sphere press has led to the enhancement of low quality natural diamonds. Synthesized and treated diamonds are sometimes traded deceptively as high quality natural diamonds because it is hard to distinguish among these diamonds with conventional gemological characterization method. Therefore, we need to develop a new identification method that is non-destructive, fast, and inexpensive. We proposed using new methods of UV fluorescence and X-ray Lang topography for checking the local HPHT stress field to distinguish these diamonds from natural ones. We observe unique differences in the local stress field images in treated diamonds using UV fluorescence and Lang topography characterization. Our result implies that our proposed methods may be appropriate for identification of the treated diamonds.

Determination of Bovine Serum Albumin by Its Enhancement Effect of Nile Blue Fluorescence

  • Lee, Sang-Hak;Suh, Jung-Kee;Li, Ming
    • Bulletin of the Korean Chemical Society
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    • v.24 no.1
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    • pp.45-48
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    • 2003
  • A novel fluorimetric method has been developed for the determination of microgram quantities of bovine serum albumin (BSA) based on its enhancement effect of Nile Blue fluorescence at 670 nm, caused by binding of Nile Blue to BSA to produce a stable water soluble complex. The binding constant of micromole Nile Blue-BSA complex was estimated by Scatchard plot method. Under the optimal conditions, the increased fluorescence intensity was linearly related to BSA concentration in the range of 0.5-12.0 ㎍/mL. The detection limit was 0.2 ㎍/mL, and the relative standard deviation of six replicate measurements was 1.4% for 10.0 ㎍/mL BSA. There was little interference from amino acids, sugars and most of metal ions.

Photoreactivity of Anthraquinones for the Analysis of Ginsenosides Using Photoreduction Fluorescence Detection-HPLC

  • Park, Man-Ki;Kim, Bak-Kwang;Park, Jeong-Hill;Shin, Young-Geun;Cho, Kyung-Hee;Do, Young-Mi
    • Archives of Pharmacal Research
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    • v.19 no.6
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    • pp.562-565
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    • 1996
  • The photoreactivity of twelve anthraquinone derivatives was examined to evaluate its usefulness as a photo-reagent for the analysis of ginsenosides using photoreduction fluorescence (PRF) detection method. Among the tested compounds, 2-tert-butylandthraquinone (TBAQ), 2-chloroanthraquinone (CAQ) and anthraquinone (AQ) showed good characteristics as photoreagents. The detection limits of ginsenoside $Rg_{1}$PRF-HPLC method using TBAQ, CAQ or AQ as a photo-reagent were found to be ca. 35 ng, 50 ng and 50 ng, respectively.

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A Simple Method for Testing Freezing Resistance Based on Chlorophyll Fluorescence in Tea (Camellia sinensis L.)

  • Chun, Jong-Un;Jeong, In-Ho;Choi, Hyoung-Kog
    • KOREAN JOURNAL OF CROP SCIENCE
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    • v.45 no.5
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    • pp.322-327
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    • 2000
  • For the stable production of high quality tea, the freezing resistance is a very important character. Most of the farmers have planted out-pollinated seeds that are not genetically pure. So, with small sample, a quick and simple method is required to test freezing resistance of lots of germ-plasm and early generation of hybrids. The absorbances(A530 nm) of TTC reduction solution at -5$^{\circ}C$ were positively correlated with resistance to photoinhibition of PSII in 6 hour photoinhibitory treatments, being significantly fitted by simple linear regression ($R^2$=${0.64}^{**}$). Chlorophyll fluorescence measured by Fv/Fm was found to be very useful in evaluating the relative levels of freezing resistance in tea.

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The Application of Time-Resolved Laser Induced Fluorescence Spectroscopy in the Complexation Studies of Eu(III) and Cm(III) with Humic Substances

  • Joong Gill Choi;Oum Ka Won;Chang Yeoul Choi;Hichung Moon;Hyun Sang Shin;Park, Seung Min;Paul Joe Chong
    • Bulletin of the Korean Chemical Society
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    • v.14 no.1
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    • pp.72-78
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    • 1993
  • The application of time-resolved laser induced fluorescence spectroscopy (TRLIF) to the complexation studies of Eu(III) and Cm(III) with humic substances is described. Using this method, three different spectroscopic characteristics(excitation spectra, emission spectra, and lifetimes) of these aquo ions and their complexes can be directly measured. By observing shifts in the wavelength and changes in the lifetime and intensities of the fluorescence emission, the information on the complexation behavior of humic substances with these trivalent metal cations in an aqueous solution, as well as energy transfer mechanisms, can be obtained. In addition, this method allows precise spectroscopic quantification of the complexation processes at very low concentrations of both components.

Analysis of DA-6034. a New Flavonoid Derivative in Biological Fluids by Fluorescence Detector

  • Jang, Ji-Myun;Park, Kyung-Jin;Lee, Jong-Jin;Kim, Dong-Goo;Shim, Hyun-Joo;Son, Mi-Won;Kim, Dong-Sung;Kim, Soon-Hoe;Yoo, Moo-Hi
    • Proceedings of the PSK Conference
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    • 2002.10a
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    • pp.403.2-403.2
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    • 2002
  • A high performance liquid chromatographic method was developed for the determination of DA-6034 in biological fluids using fluorescence detector. The method involved deproteinization of biological sample with the same volume of acetonitrile, 0.2M zinc sulphate. and 0.15M barium hydroxide. The aliquot of supernatant was injected onto Nova-pak C18 column and detected by fluorescence detector. Emission and excitation wavelength of detector were 336nm and 440nm. (omitted)

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