Development of a HPLC/MS/MS methodology for determining 3-O-methyldopa in human plasma and its application in a bioequivalence study
Rev. Inst. Adolfo Lutz (Online)
;
73(1): 96-105, jan.-mar. 2014. tab, graf
Article
in English
| LILACS, SES-SP
| ID: lil-782590
ABSTRACT
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 (8515, 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/z212.0 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 504000 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 + 50mg) in plasma samples from healthy human volunteers, following the ANVISA guidelines...
Full text:
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Index:
LILACS (Americas)
Main subject:
Plasma
/
Skatole
/
Therapeutic Equivalency
/
Chromatography, High Pressure Liquid
Limits:
Female
/
Humans
/
Male
Language:
English
Journal:
Rev. Inst. Adolfo Lutz (Online)
Year:
2014
Type:
Article
Institution/Affiliation country:
Fundação Oswaldo Cruz/BR
/
Universidade Federal do Rio de Janeiro/BR
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