Chapter title |
Quantification of Free Phenytoin by Liquid Chromatography Tandem Mass Spectrometry (LC/MS/MS)
|
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Chapter number | 25 |
Book title |
Clinical Applications of Mass Spectrometry in Drug Analysis
|
Published in |
Methods in molecular biology, January 2016
|
DOI | 10.1007/978-1-4939-3252-8_25 |
Pubmed ID | |
Book ISBNs |
978-1-4939-3251-1, 978-1-4939-3252-8
|
Authors |
Judy Peat, Clint Frazee, Uttam Garg |
Abstract |
Phenytoin (diphenylhydantoin) is an anticonvulsant drug that has been used for decades for the treatment of many types of seizures. The drug is highly protein bound and measurement of free-active form of the drug is warranted particularly in patients with conditions that can affect drug protein binding. Here, we describe a LC/MS/MS method for the measurement of free phenytoin. Free drug is separated by ultrafiltration of serum or plasma. Ultrafiltrate is treated with acetonitrile containing internal standard phenytoin d-10 to precipitate proteins. The mixture is centrifuged and supernatant is injected onto LC-MS-MS, and analyzed using multiple reaction monitoring. This method is linear from 0.1 to 4.0 μg/mL and does not demonstrate any significant ion suppression or enhancement. |
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