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Development and validation of a high‐performance liquid chromatography–electrospray ionization–MS/MS method for the simultaneous quantitation of levodopa and carbidopa in human plasma
Author(s) -
César Isabela Costa,
Byrro Ricardo Martins Duarte,
Santana e Silva Cardoso Fabiana Fernandes,
Mundim Iram Moreira,
Souza Teixeira Leonardo,
Gomes Sandro Antônio,
Bonfim Ricardo Rodrigues,
Pianetti Gerson Antônio
Publication year - 2011
Publication title -
journal of mass spectrometry
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.475
H-Index - 121
eISSN - 1096-9888
pISSN - 1076-5174
DOI - 10.1002/jms.1973
Subject(s) - chemistry , carbidopa , chromatography , electrospray ionization , protein precipitation , formic acid , electrospray , mass spectrometry , high performance liquid chromatography , selected reaction monitoring , analytical chemistry (journal) , tandem mass spectrometry , levodopa , medicine , disease , pathology , parkinson's disease
A sensitive and fast high‐performance liquid chromatography–electrospray ionization–MS/MS method for the simultaneous quantitation of levodopa and carbidopa in human plasma was developed and validated. A simple protein precipitation step with perchloric acid was used for the cleanup of plasma, and methyldopa was added as an internal standard. The analyses were carried out using an ACE C 18 column (50 × 4.6 mm i.d.; 5 µm particle size) and a mobile phase consisting of 0.2% formic acid and acetonitrile (90:10). The triple‐quadrupole mass spectrometer equipped with an electrospray source in positive mode was set up in the selective reaction monitoring mode to detect the ion transitions m/z 198.1 → m/z 107.0, m/z 227.2 → m/z 181.0, and m/z 212.1 → m/z 139.2 for levodopa, carbidopa, and methyldopa, respectively. The method was validated and proved to be linear, accurate, and precise over the range 50–5000 ng/mL for levodopa and 3–600 ng/mL for carbidopa. The proposed method was successfully applied in a pharmacokinetic study with a levodopa/carbidopa tablet formulation in healthy volunteers. Copyright © 2011 John Wiley & Sons, Ltd.