Analysis of Nucleotides by Micellar Electrokinetic Capillary Chromatography-Electrospray Ionization/Mass Spectrometry (MEKC-ESI/MS)
- Analysis of Nucleotides by Micellar Electrokinetic Capillary Chromatography-Electrospray Ionization/Mass Spectrometry (MEKC-ESI/MS)
- 샬릴 보우믹; 정봉철; 정병화
- MEKC; ESI/MS; nucleotides
- Issue Date
- The second Korea Metabolomics Society Symposium
- A method for determination of nucleotides using Micellar Electrokinetic Capillary Chromatography-Electrospray Ionization/Mass Spectrometry (MEKC-ESI/MS) is described. Extraction, separation and detection of analytes were optimized to get optimal condition of MEKC-ESI/MS for nucleotides analysis. CE was performed using an untreated silica capillary column [50 µm (i.d.) × 52 µm (o.d.) × 125 cm (length)]. Reverse polarity and selected ion monitoring (SIM) mode were applied for capillary electrophoresis (CE) and MS operation. 2 mM DTAB in 20 mM ammonium acetate (pH 3.5) selected as micellar running buffer for the best separation of analytes. 2 mM EDTA in 60% ethanol was selected for the sample preparation steps after testing the different deproteination and solid phase extraction techniques. The assay with the solvent met the commonly accepted validation limits for biological samples. LOD and LOQ in the limit of 1.0-20 μmol/L. The calibration curve was found to be linear (r2 > 0.99) for all analytes. Accuracy and precession were within ±15%. Finally, enhanced analysis of nucleotides in biological sample can be performed with the developed method.
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