Signal Enhancement of Selected Norepinephrine Metabolites Extracted from Artificial Urine Samples by Capillary Electrophoretic Separation

• Published in: International journal of molecular sciences 2024-11, Vol.25 (22), p.12227
• Main Authors: Kowalski, Piotr, Hermann, Natalia, Kroll, Dagmara, Belka, Mariusz, Bączek, Tomasz, Olędzka, Ilona
• Format: Article
• Language: English
• Subjects: Catecholamines; Chromatography; Chromatography, Micellar Electrokinetic Capillary - methods; Disease; Efficiency; Electrophoresis, Capillary - methods; Extraction (Chemistry); Health aspects; Humans; Immunoassay; Liquid-Liquid Extraction - methods; Mass spectrometry; Mental disorders; Metabolites; Methods; Methoxyhydroxyphenylglycol - analogs & derivatives; Methoxyhydroxyphenylglycol - urine; Noradrenaline; Norepinephrine - urine; Optimization; Physiological aspects; Plasma; Scientific imaging; Solid Phase Extraction - methods; Solvents; Urine; Vanilmandelic Acid - urine
• ISSN: 1422-0067; 1661-6596; 1422-0067
• DOI: 10.3390/ijms252212227

The measurement of selected norepinephrine metabolites, such as 3,4-dihydroxyphenylglycol (DHPG), 3-methoxy-4-hydroxyphenylethylenglycol (MHPG), and vanillylmandelic acid (VMA), in biological matrices-including urine-is of great clinical importance for the diagnosis and monitoring of diseases. This fact has forced researchers to evaluate new analytical methodologies for their isolation and preconcentration from biological samples. In this study, the three most popular extraction techniques-liquid-liquid extraction (LLE), solid-phase extraction (SPE), and a new 3D-printed system for dispersive solid-phase extraction (3D-DSPE)-were investigated. Micellar electrokinetic chromatography (MEKC) with a diode array detector (DAD) at 200 nm wavelength was applied to the separation of analytes, allowing for the assessment of the extraction efficiency (R) and enrichment factor (EF) for the tested extraction types. The separation buffer (BGE) consisted of 5 mM sodium tetraborate decahydrate, 50 mM SDS, 15% ( / ) MeOH, 150 mM boric acid, and 1 mM of 1-hexyl-3-methylimidazolium chloride (the apparent pH of the BGE equaled 7.3). The EF for each extraction procedure was calculated with respect to standard mixtures of the analytes at the same concentration levels. The 3D-DSPE procedure, using DVB sorbent and acetone as the desorption solvent, proved to be the most effective approach for the simultaneous extraction and determination of the chosen compounds, achieving over 3-fold signal amplification for DHPG and MHPG and over 2-fold for VMA. Moreover, all extraction protocols used for the selected norepinephrine metabolites were estimated and discussed. It was also confirmed that the 3D-DSPE-MEKC approach could be considered an effective tool for sample pretreatment and separation of chosen endogenous analytes in urine samples.