High-performance liquid chromatographic assay for catecholamines and metanephrines using fluorimetric detection with pre-column 9-fluorenylmethyloxycarbonyl chloride derivatization
| Autor: | Eric Chun Yong Chan, Paul C. Ho, P.Y Wee, P.Y Ho |
|---|---|
| Rok vydání: | 2001 |
| Předmět: |
Adult
Male Chemical ionization Fluorenes Chromatography Molecular Conformation Reproducibility of Results Atmospheric-pressure chemical ionization General Chemistry Metanephrines Mass spectrometry Normetanephrine High-performance liquid chromatography Mass Spectrometry chemistry.chemical_compound Catecholamines chemistry Humans Fluorometry Indicators and Reagents Derivatization Metanephrine Chromatography High Pressure Liquid |
| Zdroj: | Journal of chromatography. B, Biomedical sciences and applications. 749(2) |
| ISSN: | 1387-2273 |
| Popis: | A convenient HPLC-fluorescent assay of norepinephrine (NE), epinephrine (E), dopamine (DA) and their 3-O-methylated metabolites, normetanephrine (NM) and metanephrine (MN) was developed. These analytes were coupled to 9-fluorenylmethyloxycarbonyl chloride (FMOC-Cl) before assays. Results showed that using a linear gradient elution, peaks of FMOC-NE, FMOC-E, FMOC-DA, FMOC-NM, FMOC-MN and FMOC-DHBA (3,4-dihydroxybenzylamine, internal standard) were simultaneously resolved within 40 min. Optimization of the chromatographic and derivatization conditions, and validation of the assay were further discussed in the paper. The structures of these derivatives were confirmed by atmospheric pressure chemical ionization mass spectrometry (APCI-MS). The molecular ions [M+H]- of FMOC-NE, FMOC-E, FMOC-DA, FMOC-NM and FMOC-MN were m/z 836, 850, 820, 628 and 642, respectively. Based on these findings, the FMOC-derivatives of metanephrines and catecholamines were confirmed to be bi-substituted and tri-substituted respectively at the amino and catechol functional groups. Finally, the assay was successfully applied to the measurement of urinary E, DA, NM and MN after direct derivatization and simple cleaning. |
| Databáze: | OpenAIRE |
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