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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

✍ Scribed by Isabela Costa César; Ricardo Martins Duarte Byrro; Fabiana Fernandes de Santana e Silva Cardoso; Iram Moreira Mundim; Leonardo de Souza Teixeira; Sandro Antônio Gomes; Ricardo Rodrigues Bonfim; Gerson Antônio Pianetti

Publisher
John Wiley and Sons
Year
2011
Tongue
English
Weight
381 KB
Volume
46
Category
Article
ISSN
1076-5174

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✦ Synopsis

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.

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