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Comparison of capillary electrophoresis and zwitterionic-hydrophilic interaction capillary liquid chromatography with ultraviolet and mass spectrometry detection for the analysis of microRNA biomarkers

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This study evaluates zwitterionic-hydrophilic interaction capillary liquid chromatography (capZIC-HILIC) and capillary electrophoresis (CE) with ultraviolet (UV) and mass spectrometry (MS) detection for the direct, label-free and multiplex analysis of microribonucleic acids (miRNAs). CapZIC-HILIC-UV and CE-UV methods were first optimized, resulting in similar separations for a mixture of three miRNAs (hsa-iso-miR-16-5p, hsa-let-7g-5p, and hsa-miR-21-5p) but with reversal of elution/migration orders and small differences in repeatability, linearity, limit of detection (LOD) and separation efficiency. The established UV methods were transferred and validated in these terms with mass spectrometry (MS) detection, which allowed identifying the miRNAs and characterizing their post-transcriptional modifications. LOD by capZIC-HILIC-MS was 1 µM of miRNA, around 5 times lower than by CE-MS due to the analyte dilution with the sheathflow CE-MS interface and to the slightly increased abundance of alkali metals adducts in the CE-MS mass spectra. In addition, the suction effect promoted by the nebulizer gas in CE-MS negatively affected the already compromised separations. In contrast, CE-MS showed superior repeatabilities with spiked serum samples, as well as reduced costs, extended capillary column durabilities and shorter conditioning times. The comparison of the different methods allows disclosing the current advantages and disadvantages of capZIC-HILIC and CE for the analysis of miRNA biomarkers.

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Articles publicats en revistes  (Enginyeria Química i Química Analítica)

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Comparison of capillary electrophoresis and zwitterionic-hydrophilic interaction capillary liquid chromatography with ultraviolet and mass spectrometry detection for the analysis of microRNA biomarkers