title: Validation of a SPE-LC-MS/MS method for the determination of ketamine and norketamine in micropulverized hair after a single IV dose.
abstract: A SPE-LC-MS/MS method for the determination of ketamine (KET) and norketamine (NKET) was developed and validated. Extensive pulverization (25min at 25Hz) of previously cooled samples (5min in liquid nitrogen) allowed for extraction in a phosphate buffer (pH 6) solution after 10min vortex agitation at room temperature, simplifying the coupling of the extraction to an effective mixed-mode SPE (solid phase extraction) clean-up procedure. The extraction optimization was performed with samples fortified by drug incorporation according to a published procedure involving incubation of blank matrices for 16days. The method was validated for selectivity, matrix effect, linearity, LLQ (lower limit of quantification), precision, accuracy, recovery, carryover and stability after preparation and has proven to be accurate and reliable within a range of 0.02-10ng/mg for KET and 0.04-4ng/mg for NKET, meeting proposed KET cutoffs for discrimination from chronic use. In addition, the method was sensitive enough to detect the drugs after unique small (1mg/kg) intravenous doses received by patients submitted to general anesthesia before surgical procedures. Ketamine levels varied from 0.060 to 0.111ng/mg and norketamine was positive (<LLQ) in all samples. The high sensitivity achieved with conventional 3D-ion-trap MS/MS detection is probably due to the simple but very effective proposed procedure.
authors: Instituto de Química, Universidade Federal Fluminense, Outeiro de S. João Batista S/N, Niterói, RJ, Brazil.
authors: Adriana S
authors: AS
authors: Barreto
authors: Instituto de Química, Universidade Federal Fluminense, Outeiro de S. João Batista S/N, Niterói, RJ, Brazil.
authors: Vinnicius F
authors: VF
authors: Brant
authors: LaboratórioToxFree, Faculdade de Farmácia, Universidade Federal Fluminense, Rua Mário Viana, 523, Santa Rosa, Niterói, RJ, Brazil.
authors: Eliani
authors: E
authors: Spinelli
authors: Instituto de Química, Universidade Federal Fluminense, Outeiro de S. João Batista S/N, Niterói, RJ, Brazil. Electronic address: silvanavr@id.uff.br.
authors: Silvana V
authors: SV
authors: Rodrigues
mesh_terms: D000328:Adult; D000368:Aged; D002138:Calibration; D002853:Chromatography, Liquid / Q000379:methods*; D004305:Dose-Response Relationship, Drug; D005260:Female; D006197:Hair / Q000737:chemistry*; D006801:Humans; D007275:Injections, Intravenous; D007649:Ketamine / Q000008:administration & dosage* / Q000031:analogs & derivatives* / Q000032:analysis*; D057230:Limit of Detection; D015203:Reproducibility of Results; D052616:Solid Phase Extraction / Q000379:methods*; D053719:Tandem Mass Spectrometry / Q000379:methods*
chemical_list: D007649:Ketamine; C033419:norketamine
keywords: Extraction; Hair analysis; Ketamine; LC–MS/MS; Norketamine