title: Quantitation of pregabalin in dried blood spots and dried plasma spots by validated LC-MS/MS methods.
abstract: In this paper, novel LC-MS/MS methods for the determination of antiepileptic drug pregabalin in dried matrix spots (DMS) are presented. This attractive technique of sample collection in micro amount was utilized in the form of dried blood spots (DBS) and dried plasma spots (DPS). Following a pre-column derivatization procedure, using n-propyl chloroformate in the presence of n-propanol, and consecutive liquid-liquid extraction, derivatized pregabalin and its internal standard, 4-aminocyclohexanecarboxylic acid, were detected in positive ion mode by applying two SRM transitions per analyte. A YMC-Pack Octyl column (50mm×4.0mm, 3μm particle size) maintained at 30°C, was utilized with running mobile phase composed of acetonitrile: 0.15% formic acid (85:15, v/v). Flow rate was 550μL/min and total run time 2min. Established methods were fully validated over the concentration range of 0.200-20.0μg/mL for DBS and 0.400-40.0μg/mL for DPS, respectively, while specificity, accuracy, precision, recovery, matrix-effect, stability, dilution integrity and spot homogeneity were found within acceptance criteria. Validated methods were applied for the determination of pregabalin levels in dried blood and plasma samples obtained from patients with epilepsy, after per os administration of commercial capsules. Comparison of drug level in blood and plasma, as well as correction steps undertaken in order to overcome hematocrit issue, when analyzing DBS, are also given.
authors.affiliation: University of Belgrade, Faculty of Pharmacy, Department of Drug Analysis, Vojvode Stepe 450, Belgrade, Serbia.
authors.forename: Nađa
authors.initials: N
authors.lastname: Kostić
authors.affiliation: University of Athens, School of Pharmacy, Department of Pharmaceutical Chemistry, Panepistimioupoli Zografou, Athens, Greece.
authors.forename: Yannis
authors.initials: Y
authors.lastname: Dotsikas
authors.affiliation: University of Belgrade, Faculty of Medicine, Clinic of Neurology and Psychiatry for Children and Youth, Dr Subotića 6a, Belgrade, Serbia.
authors.forename: Nebojša
authors.initials: N
authors.lastname: Jović
authors.affiliation: University of Belgrade, Faculty of Medicine, Clinic of Neurology and Psychiatry for Children and Youth, Dr Subotića 6a, Belgrade, Serbia.
authors.forename: Galina
authors.initials: G
authors.lastname: Stevanović
authors.affiliation: University of Belgrade, Faculty of Pharmacy, Department of Drug Analysis, Vojvode Stepe 450, Belgrade, Serbia. Electronic address: andja@pharmacy.bg.ac.rs.
authors.forename: Anđelija
authors.initials: A
authors.lastname: Malenović
authors.affiliation: University of Belgrade, Faculty of Pharmacy, Department of Physical Chemistry and Instrumental Methods, Vojvode Stepe 450, Belgrade, Serbia.
authors.forename: Mirjana
authors.initials: M
authors.lastname: Medenica
mesh_terms: D000293:Adolescent; D000328:Adult; D000927:Anticonvulsants / Q000097:blood*; D002138:Calibration; D002648:Child; D002851:Chromatography, High Pressure Liquid; D005260:Female; D006400:Hematocrit; D006801:Humans; D008297:Male; D013058:Mass Spectrometry; D000069583:Pregabalin / Q000097:blood*; D011786:Quality Control; D012015:Reference Standards; D015203:Reproducibility of Results; D055815:Young Adult
chemical_list: D000927:Anticonvulsants; D000069583:Pregabalin
keywords: Chloroformates; Dried blood/plasma spots; Hematocrit; Mass spectrometry; Pregabalin