Enzymatic determination of ethanol based on the intrinsic fluorescence of alcohol dehydrogenase

被引:22
作者
deMarcos, S
Galban, J
Albajez, R
Castillo, JR
机构
[1] Analyt. Spectrosc. and Sensors Grp., Analytical Department, University of Zaragoza
关键词
alcohol dehydrogenase; intrinsic fluorescence; ethanol and drink samples;
D O I
10.1016/S0003-2670(96)00617-4
中图分类号
O65 [分析化学];
学科分类号
070302 ; 081704 ;
摘要
A fluorimetric enzymatic method for the determination of ethanol is presented. The method is based on the intrinsic fluorescence changes caused in alcohol dehydrogenase (ADH) during the enzymatic reaction. This Variation is due to the different inner filter effects of the co-enzyme (NAD(+)) and its reduced form (NADH). The latter is generated during the enzymatic reaction. A study is made of the origin of the ADH fluorescence signal using a mathematical model, which makes it possible to explain the linear response between the enzyme's analytical signal and the concentration of ethanol ranging from 5.5 x 10(-5) to 3.2 x 10(-3) M. The 3 sigma limit of detection is 3.1 x 10(-6) M, and the relative standard deviation for 6 determinations at 1.0 x 10(-4) M ethanol is 2.8%. The method is applied to low ethanol concentration drink samples and vinegar without pre-treatment.
引用
收藏
页码:117 / 123
页数:7
相关论文
共 19 条
  • [1] QUANTITATIVE-DETERMINATION OF ORGANIC-SOLVENTS BY CAPILLARY ELECTROPHORESIS USING INDIRECT UV DETECTION
    ALTRIA, KD
    HOWELLS, JS
    [J]. JOURNAL OF CHROMATOGRAPHY A, 1995, 696 (02) : 341 - 348
  • [2] GAS-CHROMATOGRAPHY MASS-SPECTROMETRY ANALYSIS OF VOLATILE ORGANIC-COMPOUNDS PRODUCED BY SOME MICROMYCETES
    CAILLEUX, A
    BOUCHARA, JP
    DANIEL, V
    CHABASSE, D
    ALLAIN, P
    [J]. CHROMATOGRAPHIA, 1992, 34 (11-12) : 613 - 617
  • [3] DEMARCOS S, 1995, ANAL SCI, V11, P233, DOI 10.2116/analsci.11.233
  • [4] ENZYMATIC DETERMINATION OF ALCOHOL MIXTURES AT THE NANOGRAM LEVEL BY THE STOPPED-FLOW TECHNIQUE
    FORSTER, E
    SILVA, M
    OTTO, M
    PEREZBENDITO, D
    [J]. ANALYTICA CHIMICA ACTA, 1993, 274 (01) : 109 - 116
  • [5] DETERMINATION OF LACTATE BY THE INTRINSIC FLUORESCENCE OF LACTATE OXIDASE
    GALBAN, J
    DEMARCOS, S
    SEGURA, P
    CASTILLO, JR
    [J]. ANALYTICA CHIMICA ACTA, 1994, 299 (02) : 277 - 284
  • [6] FLUOROMETRIC ENZYMATIC LACTATE DETERMINATION BASED ON ENZYME CYTOCHROME-B(2) FLUORESCENCE
    GALBAN, J
    DEMARCOS, S
    CASTILLO, JR
    [J]. ANALYTICAL CHEMISTRY, 1993, 65 (21) : 3076 - 3080
  • [7] DERIVATIVE FOURIER-TRANSFORM INFRARED SPECTROMETRIC DETERMINATION OF ETHANOL IN BEERS
    GALLIGNANI, M
    GARRIGUES, S
    DELAGUARDIA, M
    [J]. ANALYST, 1994, 119 (08) : 1773 - 1778
  • [8] ETHANOL BIOSENSOR USING IMMOBILIZED COENZYME
    GOTOH, M
    KARUBE, I
    [J]. ANALYTICAL LETTERS, 1994, 27 (02) : 273 - 284
  • [9] An amperometric aqueous ethanol sensor based on the electrocatalytic oxidation at a cobalt-nickel oxide electrode
    Hayes, ET
    Bellingham, BK
    Mark, HB
    Galal, A
    [J]. ELECTROCHIMICA ACTA, 1996, 41 (02) : 337 - 344
  • [10] LAKOWICK JR, 1991, TOPICS FLUORESCENCE, V2, pCH3