A Specific Method for the Measurement of Cyclosporin A in Human Whole Blood by Liquid Chromatography-Tandem Mass Spectrometry

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Abstract
Therapeutic monitoring of the immunosuppressant cyclosporin A (CsA) is routinely performed by immunoassays to make individual dosage adjustments for patients after organ transplantation. High-performance liquid chromatography with ultraviolet detection (HPLC-UV) has been used as the reference method. However, HPLC-UV methods frequently suffer from chromatographic interferences that affect accuracy and reproducibility. A sensitive, specific HPLC-tandem mass spectrometry (HPLC/MS/MS) method for the quantitation of CsA has been developed. One hundred microliters CsA whole blood sample containing cyclosporin C (CsC) as the internal standard was extracted with ethyl ether. High-performance liquid chromatography separation was accomplished on an RP-C18 narrow-bore column at 50°C with a linear gradient elution followed by on-line ion-spray ionization MS/MS analysis. The standard curve was established in the range of 10 to 1000 µg/l (r = 0.9989,n = 8). Limits of detection and quantitation were 1 µg/l and 5 µg/l, respectively. Imprecision was r

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