An analysis of avidin, biotin and their interaction at attomole levels by voltammetric and chromatographic techniques
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- 作者：Rene Kizek ; Michal Masarik ; Karl J. Kramer ; David Potesil ; Michele Bailey ; John A. Howard ; Borivoj Klejdus ; Radka Mikelova ; Vojtech Adam and Libuse Trnkova ; et al.
- 关键词：Avidin ; Biotin ; Avidin– ; biotin technology ; Square ; wave voltammetry ; Modified electrode ; High ; performance liquid chromatography ; Diode array detection ; Electrochemical transfer technique ; Carbon paste electrode ; Drug analysis
- 刊名：Analytical and Bioanalytical Chemistry
- 出版年：2005
- 出版时间：March 2005
- 年：2005
- 卷：381
- 期：6
- 页码：1167-1178
- 全文大小：602 KB
- 刊物类别：Chemistry and Materials Science
- 刊物主题：Chemistry
Analytical Chemistry
Food Science
Inorganic Chemistry
Physical Chemistry
Monitoring, Environmental Analysis and Environmental Ecotoxicology - 出版者：Springer Berlin / Heidelberg
- ISSN：1618-2650
文摘
The electroanalytical determination of avidin in solution, in a carbon paste, and in a transgenic maize extract was performed in acidic medium at a carbon paste electrode (CPE). The oxidative voltammetric signal resulting from the presence of tyrosine and tryptophan in avidin was observed using square-wave voltammetry. The process could be used to determine avidin concentrations up to 3 fM (100 amol in 3 l drop) in solution, 700 fM (174 fmol in 250 l solution) in an avidin-modified electrode, and 174 nM in a maize seed extract. In the case of the avidin-modified CPE, several parameters were studied in order to optimize the measurements, such as electrode accumulation time, composition of the avidin-modified CPE, and the elution time of avidin. In addition, the avidin-modified electrode was used to detect biotin in solution (the detection limit was 7.6 pmol in a 6 l drop) and to detect biotin in a pharmaceutical drug after various solvent extraction procedures. Comparable studies for the detection of biotin were developed using HPLC with diode array detection (HPLC-DAD) and flow injection analysis with electrochemical detection, which allowed biotin to be detected at levels as low as 614 pM and 6.6 nM, respectively. The effects of applied potential, acetonitrile content, and flow rate of the mobile phase on the FIA-ED signal were also studied.