Quantification of perifosine, an alkylphosphocholine anti-tumour agent, in plasma by pneumatically assisted electrospray tandem mass spectrometry coupled with high-performance liquid chromatography.
An HPLC assay with tandem mass spectrometric detection in the positive-ion Turbo-Ion-Spray (TISP) mode for the fast and sensitive determination of perifosine ((I), D-21266) in human plasma was developed, utilising the structural analogue, miltefosine ((II), D-18506), as internal standard. Automated solid-phase extraction of diluted plasma samples, based on 250-microl plasma aliquots, at pH 6.5, allowed a reliable quantification of perifosine down to 4 ng/ml. Injection of 200 microl of plasma extracts onto a 100x3 mm normal-phase analytical column at a flow-rate of 0.5 ml/min provided retention-times of 2.4 and 2.1 min for perifosine (I) and the internal standard (II), respectively. The standard curves were linear from 4 to 2000 ng/ml using weighted linear regression analysis (1/Y2). The inter-assay and intra-assay accuracies for the calibration standards were within +0.9% and -0.2%, exhibiting precisions (C.V.) of +/-6.5 and +/-7.3%, respectively. Up to 100 unknowns may be analysed each 24 h per analyst.
Department of Biological Research Biochemistry, ASTA Medica AG, Frankfurt, Germany. Dr_Norbert.Knebel@astamedica.de
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Knebel
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Grieb
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Winkler
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Locher
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van der Vlis
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Verheij
D002138:Calibration; D002851:Chromatography, High Pressure Liquid; D006801:Humans; D013058:Mass Spectrometry; D010767:Phosphorylcholine / Q000031:analogs & derivatives* / Q000097:blood / Q000493:pharmacokinetics; D011786:Quality Control; D013050:Spectrometry, Fluorescence; D013056:Spectrophotometry, Ultraviolet
D010767:Phosphorylcholine; C105905:perifosine