Microscale high-performance liquid chromatography–electrospray tandem mass spectrometry assay for cyclosporin A in blood |
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| Authors: | Paul J Taylor Christopher E Jones Paul T Martin Stephen V Lynch Anthony G Johnson Susan M Pond |
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| Institution: | aDepartment of Clinical Pharmacology, First Floor Lions Clinical Research Building, Princess Alexandra Hospital, Ipswich Road, Brisbane, QLD 4102, Australia;bUniversity of Queensland Department of Medicine, Princess Alexandra Hospital, Brisbane, QLD 4102, Australia;cUniversity of Queensland Department of Surgery, Princess Alexandra Hospital, Brisbane, QLD 4102, Australia;dJohnson and Johnson Research, Sydney, NSW 2065, Australia |
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| Abstract: | To facilitate quantitative analysis of cyclosporin A in low volume blood samples we developed a sensitive and specific microscale reversed-phase HPLC–electrospray tandem mass spectrometry assay. Blood samples (100 μl) were prepared by acetonitrile precipitation and C18 solid-phase extraction. Detection was by multiple-reactant monitoring. The method was linear over the range 5–1000 μg/l (r≥0.997) with accuracy between 95.4 and 102.0% over this range. Total imprecision was 11.1% at 10 μg/l and 2.8% at 800 μg/l. Absolute recovery of cyclosporin A and internal standard was 72.5 and 73.3%, respectively. When this method was evaluated against a conventional HPLC with UV detection, in patient samples, they were interchangeable (y=0.988x+10.0, r=0.996). This HPLC–ESI-MS–MS method will be applicable to therapeutic monitoring in paediatric transplant patients and multiple point pharmacokinetic studies in animals and humans. |
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| Keywords: | Cyclosporin A |
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