title: Quantitative determination of fentanyl in newborn pig plasma and cerebrospinal fluid samples by HPLC-MS/MS.
abstract: In this study, a selective and sensitive high performance liquid chromatography-tandem mass spectrometry (HPLC-MS/MS) method requiring low sample volume (≤100 μL) was developed and validated for the quantitative determination of the opioid drug fentanyl in plasma and cerebrospinal fluid (CSF). A protein precipitation extraction with acetonitrile was used for plasma samples whereas CSF samples were injected directly on the HPLC column. Fentanyl and (13) C6 -fentanyl (Internal Standard) were analyzed in an electrospray ionization source in positive mode, with multiple reaction monitoring (MRM) of the transitions m/z 337.0/188.0 and m/z 337.0/105.0 for quantification and confirmation of fentanyl, and m/z 343.0/188.0 for (13) C6 -fentanyl. The respective lowest limits of quantification for plasma and CSF were 0.2 and 0.25 ng/mL. Intra- and inter-assay precision and accuracy did not exceed 15%, in accordance with bioanalytical validation guidelines. The described analytical method was proven to be robust and was successfully applied to the determination of fentanyl in plasma and CSF samples from a pharmacokinetic and pharmacodynamic study in newborn piglets receiving intravenous fentanyl (5 µg/kg bolus immediately followed by a 90-min infusion of 3 µg/kg/h).
authors.affiliation: Analytical Chemistry Department, Science and Technology Faculty, University of the Basque Country (UPV/EHU), Bilbao, Basque Country, Spain.
authors.forename: M E
authors.initials: ME
authors.lastname: Blanco
authors.affiliation: Pharmacology Department, Faculty of Medicine, University of the Basque Country (UPV/EHU), Bilbao, Basque Country, Spain.
authors.forename: E
authors.initials: E
authors.lastname: Encinas
authors.affiliation: Analytical Chemistry Department, Science and Technology Faculty, University of the Basque Country (UPV/EHU), Bilbao, Basque Country, Spain.|Analytical Bioscience Division, LACDR, Leiden University, Leiden, the Netherlands.
authors.forename: O
authors.initials: O
authors.lastname: González
authors.affiliation: Analytical Chemistry Department, Science and Technology Faculty, University of the Basque Country (UPV/EHU), Bilbao, Basque Country, Spain.
authors.forename: E
authors.initials: E
authors.lastname: Rico
authors.affiliation: Drug Modeling & Consulting, Dynakin, SL, Bilbao, Basque Country, Spain.
authors.forename: V
authors.initials: V
authors.lastname: Vozmediano
authors.affiliation: Pharmacology Department, Faculty of Medicine, University of the Basque Country (UPV/EHU), Bilbao, Basque Country, Spain.
authors.forename: E
authors.initials: E
authors.lastname: Suárez
authors.affiliation: Analytical Chemistry Department, Science and Technology Faculty, University of the Basque Country (UPV/EHU), Bilbao, Basque Country, Spain.
authors.forename: R M
authors.initials: RM
authors.lastname: Alonso
mesh_terms: D000701:Analgesics, Opioid / Q000008:administration & dosage / Q000097:blood* / Q000134:cerebrospinal fluid*; D000818:Animals; D000831:Animals, Newborn; D002851:Chromatography, High Pressure Liquid / Q000379:methods*; D005283:Fentanyl / Q000008:administration & dosage / Q000097:blood* / Q000134:cerebrospinal fluid*; D057230:Limit of Detection; D015203:Reproducibility of Results; D021241:Spectrometry, Mass, Electrospray Ionization / Q000379:methods; D013552:Swine; D053719:Tandem Mass Spectrometry / Q000379:methods*
chemical_list: D000701:Analgesics, Opioid; D005283:Fentanyl
keywords: HPLC-MS/MS; cerebrospinal fluid; fentanyl; newborn pig; plasma