Dried blood spot assay for the quantification of phenytoin using Liquid Chromatography-Mass Spectrometry

Fabio Villanelli, Elisa Giocaliere, Sabrina Malvagia, Anna Rosati, Giulia Forni, Silvia Funghini, Engy Shokry, Daniela Ombrone, Maria Luisa Della Bona, Renzo Guerrini, Giancarlo la Marca

Research output: Contribution to journalArticlepeer-review


Phenytoin (PHT) is one of the most commonly used anticonvulsant drugs for the treatment of epilepsy and bipolar disorders.The large amount of plasma required by conventional methods for drug quantification makes mass spectrometry combined with dried blood spot (DBS) sampling crucial for pediatric patients where therapeutic drug monitoring or pharmacokinetic studies may be difficult to realize. DBS represents a new convenient sampling support requiring minimally invasive blood drawing and providing long-term stability of samples and less expensive shipment and storage.The aim of this study was to develop a LC-MS/MS method for the quantification of PHT on DBS.This analytical method was validated and gave good linearity (r2=0.999) in the range of 0-100mg/l. LOQ and LOD were 1.0mg/l and 0.3mg/l, respectively.The drug extraction from paper was performed in a few minutes using a mixture composed of organic solvent for 80%. The recovery ranged from 85 to 90%; PHT in DBS showed to be stable at different storage temperatures for one month. A good correlation was also obtained between PHT plasma and DBS concentrations.This method is both precise and accurate and appears to be particularly suitable to monitor treatment with a simple and convenient sample collection procedure.

Original languageEnglish
Pages (from-to)31-35
Number of pages5
JournalClinica Chimica Acta
Publication statusPublished - Feb 2 2015


  • Anticonvulsant drug
  • Dried blood spot
  • Epilepsy
  • Phenytoin
  • Tandem mass spectrometry
  • Therapeutic drug monitoring

ASJC Scopus subject areas

  • Biochemistry
  • Clinical Biochemistry
  • Biochemistry, medical
  • Medicine(all)


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