FLUOROMETRIC ENZYMATIC LACTATE DETERMINATION BASED ON ENZYME CYTOCHROME-B(2) FLUORESCENCE

被引:28
作者
GALBAN, J
DEMARCOS, S
CASTILLO, JR
机构
[1] Department of Analytical Chemistry, Faculty of Sciences, University of Zaragoza, Zaragoza-50009, Pza. San Francisco s/n
关键词
D O I
10.1021/ac00069a022
中图分类号
O65 [分析化学];
学科分类号
070302 ; 081704 ;
摘要
This paper presents a procedure for fluorometric-enzymatic lactate determination based on the modification of the fluorometric properties of the enzyme L-lactic dehydrogenase (cytochrome b2), during the enzymatic oxidation of the analyte with ferricyanide. During the reaction one can observe an irreversible fall in the intensity of the enzyme's fluorescence, the rate of which is proportional to the concentration of the lactate. The source of this signal has been investigated and it has been shown that, besides the formation of a complex between the enzyme and the ferricyanide (the constant of which can be determined), this signal loss can be explained by simultaneous inner filter effects caused by the ferricyanide and the ferrocyanide (generated in the enzymatic reaction). A mathematical model has been developed which makes it possible to establish a linear response between the enzyme's analytical signal of fluorescence and the concentrations of the lactate, the cytochrome, and the ferricyanide. The procedure makes it possible to determine the lactate in concentrations ranging from 0.2 to 45 mg/L. Determination of the analyte has been carried out in milk samples with great precision and accuracy.
引用
收藏
页码:3076 / 3080
页数:5
相关论文
共 19 条
[1]   FLOW-INJECTION ANALYSIS OF MICROMOLAR CONCENTRATIONS OF GLUCOSE AND LACTATE IN FERMENTATION MEDIA [J].
BENTHIN, S ;
NIELSEN, J ;
VILLADSEN, J .
ANALYTICA CHIMICA ACTA, 1992, 261 (1-2) :145-153
[2]  
BICHSEL S, 1984, PHARM ACTA HELV, V59, P62
[3]  
CASTILLO JR, IN PRESS SENS ACTUAT
[4]  
Dixon M., 1979, ENZYMES, P47
[5]   REAGENTLESS AMPEROMETRIC LACTATE ELECTRODE [J].
DURLIAT, H ;
COMTAT, M .
ANALYTICAL CHEMISTRY, 1980, 52 (13) :2109-2112
[6]   SPECTROPHOTOMETRIC METHOD FOR THE DETERMINATION OF MICROQUANTITIES OF LACTIC-ACID IN BIOLOGICAL-MATERIAL [J].
FIGENSCHOU, DL ;
MARAIS, JP .
ANALYTICAL BIOCHEMISTRY, 1991, 195 (02) :308-312
[7]  
GAFFNEY M, 1984, ANAL P, V21, P434
[8]   FLUOROMETRIC-DETERMINATION OF CADMIUM IN POLYVINYL-CHLORIDE STABILIZERS AND POLYVINYL-CHLORIDE IN NONAQUEOUS MEDIA [J].
GALBAN, J ;
DEMARCOS, S ;
VIDAL, JC ;
DIAZ, C ;
AZNAREZ, J .
ANALYTICAL SCIENCES, 1990, 6 (02) :187-190
[9]   EFFECTS OF ABSORPTION AND SELF-ABSORPTION QUENCHING ON FLUORESCENT INTENSITIES [J].
HENDERSON, G .
JOURNAL OF CHEMICAL EDUCATION, 1977, 54 (01) :57-59
[10]   FLOW-INJECTION METHOD FOR THE POLAROGRAPHIC-DETERMINATION OF L-LACTATE USING AN IMMOBILIZED ENZYME REACTOR [J].
HIKIMA, S ;
HASEBE, K ;
KAKIZAKI, T .
ANALYTICAL SCIENCES, 1992, 8 (02) :165-168