Development of a HPLC/MS/MS methodology for determining 3-O-methyldopa in human plasma and its application in a bioequivalence study
| Autor: | Fábio Coelho Amendoeira, Heliana Figueiredo Martins, Marlice Aparecida Sipoli Marques, Viviane de Assis Nascimento, Douglas Pereira Pinto |
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| Rok vydání: | 2000 |
| Předmět: |
Analyte
Chromatography Formic acid Calibration curve Mechanical Engineering Extraction (chemistry) Selected reaction monitoring Analytical chemistry Energy Engineering and Power Technology Management Science and Operations Research Mass spectrometry High-performance liquid chromatography Triple quadrupole mass spectrometer chemistry.chemical_compound chemistry |
| Zdroj: | Revista do Instituto Adolfo Lutz. |
| ISSN: | 1983-3814 0073-9855 |
| DOI: | 10.18241/0073-98552014731593 |
| Popis: | A simple, sensitive and specific HPLC/MS/MS methodology was developed and it was validated for determining 3-O-methyldopa, the major metabolite of dopamine, in human plasma. The separation was achieved on Atlantis T3 C18 analytical column (5 μm; 150 x 4.6 mm i.d.) using a mobile phase consisted of a solution of water and methanol (85:15, v/v) and containing formic acid 0.05%. The extraction from the analyte and the internal standard sample was performed using a simple protein plasma precipitation with perchloric acid. The detection was conducted on a triple quadrupole tandem mass spectrometer with a positive multiple reaction monitoring mode (MRM). The monitored fragmentation transitions were m/z 212.0 - m/z 166.0 for 3-O-methyldopa and m/z 227.10 - m/z 181.0 for carbidopa (internal standard). The calibration curves were linear in the range of 50–4000 ng/mL for 3-O-methyldopa. The methodology presented a good precision and accuracy in accordance to the criteria for biomedical analysis. And it was successfully applied to the bioequivalence study of two formulations levodopa + benserazide (200 + 50 mg) in plasma samples from healthy human volunteers, following the ANVISA guidelines. |
| Databáze: | OpenAIRE |
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