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標題: Development of an enzymatic chromatography strip with nicotinamide adenine dinucleotide-tetrazolium coupling reactions for quantitative l-lactate analysis
作者: Kan, Shu-Chen
Chang, Wei-Feng
Lan, Min-Chi
Lin, Chia-Chi
Lai, Wei-Shiang
Shieh, Chwen-Jen
Hsiung, Kuang-Pin
Liu, Yung-Chuan
關鍵字: Chromatographic strip;Point-of-care testing;Tetrazolium salts;l-Lactate;l-Lactate dehydrogenase;Animals;Chromatography;Clostridium kluyveri;Enzymes, Immobilized;Humans;Hydrogen-Ion Concentration;Kinetics;L-Lactate Dehydrogenase;Lactic Acid;Limit of Detection;NAD;Rabbits;Reagent Strips;Tetrazolium Salts
Project: Analytical biochemistry, Volume 471, Page(s) 61-6.
In this study, a dry assay of l-lactate via the enzymatic chromatographic test (ECT) was developed. An l-lactate dehydrogenase plus a nicotinamide adenine dinucleotide (NADH) regeneration reaction were applied simultaneously. Various tetrazolium salts were screened to reveal visible color intensities capable of determining the lactate concentrations in the sample. The optimal analysis conditions were as follows. The diaphorase (0.5 μl, 2(-6)U/μl) was immobilized in the test line of the ECT strip. Nitrotetrazolium blue chloride (5 μl, 12 mM), l-lactate dehydrogenase (1 μl, 0.25U/μl), and NAD(+) (2μl, 1.5×10(-5)M) were added into the mobile phase (100 μl) composed of 0.1% (w/w) Tween 20 in 10mM phosphate buffer (pH 9.0), and the process was left to run for 10 min. This detection had a linear range of 0.039 to 5mM with a detection limit of 0.047 mM. This quantitative analysis process for l-lactate was easy to operate with good stability and was proper for the point-of-care testing applications.
DOI: 10.1016/j.ab.2014.11.015
Appears in Collections:化學工程學系所

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