Determination of -Carnitine by Flow Injection Analysis with NADH Fluorescence Detection
| Autor: | J.L. Iborra, Arturo Manjón, J.M. Obon |
|---|---|
| Rok vydání: | 2000 |
| Předmět: |
Biophysics
Sensitivity and Specificity Biochemistry Carnitine medicine Nadh fluorescence Molecular Biology Flow injection analysis Detection limit chemistry.chemical_classification Chromatography Chemistry Carnitine dehydrogenase Reproducibility of Results Cell Biology Enzymes Immobilized NAD Fluorescence Alcohol Oxidoreductases Spectrometry Fluorescence Enzyme Pharmaceutical Preparations Calibration Flow Injection Analysis medicine.drug |
| Zdroj: | Analytical Biochemistry. 281:176-181 |
| ISSN: | 0003-2697 |
| DOI: | 10.1006/abio.2000.4573 |
| Popis: | A flow injection analysis method for determining l -carnitine is reported. The system uses the enzyme l -carnitine dehydrogenase covalently immobilized to Eupergit C. The NADH produced by the action of the enzyme, which is proportional to the l -carnitine concentration, is quantified using fluorescence detection. The system response was rapid and had a wide range of linearity. At a flow rate of 0.2 ml/min, a detection limit of 1 μM (20 pmol) was obtained for l -carnitine, peak areas were linear up to 100 μM, and samples could be injected every 4 min. The method performed well as a routine assay, showing high sensitivity (54,000 AU/μM), a precision of 0.96%, and the ability to carry out 144 consecutive assays with an RSD of 1.47% (good stability). Comparisons were made with other known methods for l -carnitine determination. Presence of d -carnitine had no effect on l -carnitine assay. The analysis was valid for determining l -carnitine concentrations in commercial pharmaceutical preparations. |
| Databáze: | OpenAIRE |
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