title: Development and validation of a sensitive UHPLC-MS/MS method for the simultaneous analysis of tramadol, dextromethorphan chlorpheniramine and their major metabolites in human plasma in forensic context: application to pharmacokinetics.
abstract: The prerequisites for forensic confirmatory analysis by LC/MS/MS with respect to European Union guidelines are chromatographic separation, a minimum number of two MS/MS transitions to obtain the required identification points and predefined thresholds for the variability of the relative intensities of the MS/MS transitions (MRM transitions) in samples and reference standards. In the present study, a fast, sensitive and robust method to quantify tramadol, chlorpheniramine, dextromethorphan and their major metabolites, O-desmethyltramadol, dsmethyl-chlorpheniramine and dextrophan, respectively, in human plasma using ibuprofen as internal standard (IS) is described. The analytes and the IS were extracted from plasma by a liquid-liquid extraction method using ethyl acetate-diethyl-ether (1:1). Extracted samples were analyzed by ultra-high-performance liquid chromatography coupled to electrospray ionization tandem mass spectrometry (UHPLC-ESI-MS/MS). Chromatographic separation was performed by pumping the mobile phase containing acetonitrile, water and formic acid (89.2:11.7:0.1) for 2.0 min at a flow rate of 0.25 μL/min into a Hypersil-Gold C18 column, 20 × 2.0 mm (1.9 µm) from Thermoscientific, New York, USA. The calibration curve was linear for the six analytes. The intraday precision (RSD) and accuracy (RE) of the method were 3-9.8 and -1.7-4.5%, respectively. The analytical procedure herein described was used to assess the pharmacokinetics of the analytes in 24 healthy volunteers after a single oral dose containing 50 mg of tramadol hydrochloride, 3 mg chlorpheniramine maleate and 15 mg of dextromethorphan hydrobromide.
authors.affiliation: Analytical Toxicology Laboratory, Forensic Medicine Authority, Ismailia, Egypt.
authors.forename: Hala M
authors.initials: HM
authors.lastname: Heneedak
authors.affiliation: Department of Pharmaceutical Chemistry, Faculty of Pharmacy, Suez Canal University, Ismailia, Egypt.
authors.forename: Ismail
authors.initials: I
authors.lastname: Salama
authors.affiliation: Department of Pharmaceutical Chemistry, Faculty of Pharmacy, Suez Canal University, Ismailia, Egypt.
authors.forename: Samia
authors.initials: S
authors.lastname: Mostafa
authors.affiliation: Department of Pharmaceutical Chemistry, Faculty of Pharmacy, Cairo University, Cairo, Egypt.
authors.forename: Ehab
authors.initials: E
authors.lastname: El-Kady
authors.affiliation: Department of Medicinal Chemistry, Faculty of Pharmacy, Zagazig University, Zagazig, Egypt.
authors.forename: Mohamed
authors.initials: M
authors.lastname: El-Sadek
mesh_terms: D002744:Chlorpheniramine / Q000097:blood* / Q000737:chemistry / Q000493:pharmacokinetics; D002851:Chromatography, High Pressure Liquid / Q000379:methods*; D003915:Dextromethorphan / Q000097:blood* / Q000737:chemistry / Q000493:pharmacokinetics; D004355:Drug Stability; D006801:Humans; D057230:Limit of Detection; D016014:Linear Models; D015203:Reproducibility of Results; D053719:Tandem Mass Spectrometry / Q000379:methods*; D014147:Tramadol / Q000097:blood* / Q000737:chemistry / Q000493:pharmacokinetics
chemical_list: D014147:Tramadol; D002744:Chlorpheniramine; D003915:Dextromethorphan
keywords: LC-MS/MS; chlorpheniramine; dextromethorphan; plasma; tramadol