Enantioselective quantitative analysis of amphetamine in human plasma by liquid chromatography/high-resolution mass spectrometry.
A method for the quantitative enantioselective analysis of amphetamine in human plasma by LC-HRMS is presented. High-resolution detection, alone and in combination with targeted MS/MS, was validated and compared to a highly sensitive GC-NICI-Method. Derivatization with (S)-N-(heptafluorobutyryl)-prolyl chloride was accomplished to yield derivatives suitable for enantioselective analysis of amphetamine on a nonchiral reversed phase column with MS-compatible mobile phase. Equal analytical performance was observed for the methods presented and the GC-NICI-MS method. A dynamic range of 4,000 was found for the established calibration curves. A fivefold deuterated analogue of both enantiomeres was used as an internal standard. Full validation data are given to demonstrate the usefulness of the assay, including specificity, linearity, accuracy and precision, autosampler stability, matrix effect, and prospective analytical batch size accuracy. The method has been successfully applied to pharmacokinetic profiling of the drug after oral application.
Department of Pediatrics, Research Unit of Analytical Mass Spectrometry, Cell Biology and Biochemistry of Inborn Errors of Metabolism, Medical University of Graz, Auenbruggerplatz 34/2, 8036, Graz, Austria, hans.leis@medunigraz.at.
Hans J
HJ
Leis
Günter
G
Fauler
Werner
W
Windischhofer
D000284:Administration, Oral; D000661:Amphetamine / Q000008:administration & dosage / Q000097:blood* / Q000737:chemistry / Q000493:pharmacokinetics; D002138:Calibration; D002853:Chromatography, Liquid / Q000379:methods*; D005466:Fluorocarbons / Q000737:chemistry; D006801:Humans; D011392:Proline / Q000031:analogs & derivatives / Q000737:chemistry; D015203:Reproducibility of Results; D012680:Sensitivity and Specificity; D013237:Stereoisomerism; D053719:Tandem Mass Spectrometry / Q000379:methods*
D005466:Fluorocarbons; C058908:N-heptafluorobutyrylproline; D011392:Proline; D000661:Amphetamine