Cyclosporin A, Tacrolimus and Sirolimus are routinely measured in whole blood to monitor immunosuppressive therapy in transplant patients. Automating the procedure offers several advantages over the manual process, including a reduction in health and safety concerns for analysts. This work outlines a fully automated method for immunosuppressant analysis featuring automated, internal standard and reagent addition, protein precipitation and centrifugation. An optional solid phase extraction clean-up stage was developed and validated. The system is capable of handling 98 samples in under 9 hours (extraction procedure without SPE). If cleaner extracts are required, the SPE method offers greater levels of sensitivity and a batch of 98 samples can still be prepared in 16 hours and 33 minutes. Despite the increase in sample preparation time, the benefits of sensitivity and reduced potential for repeat analysis offsets the negative impacts on preparation time.
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Automating the analysis of selected Immunosuppressants in whole blood using a GERSTEL MultiPurpose Sampler coupled to liquid chromatography tandem mass spectrometry detection