Quantitative analysis of sirolimus (Rapamycin) in blood by high-performance liquid chromatography-electrospray tandem...

We report here a quantitative method for the analysis of sirolimus in blood using solid-phase sample preparation and HPLC-electrospray-tandem mass spectrometry detection. Blood samples (500 microl) were prepared by pre-treatment with acetonitrile: 15 mM zinc sulphate (70:30, v/v), containing 32-demethoxysirolimus (internal standard) and C18 solid-phase extraction. The electrospray conditions were chosen to enhance the [M+NH4]+ species at the expense of other species. Detection was by multiple reactant monitoring with the mass transitions m/z 931.8-->864.6 and m/z 901.8-->834.4 employed for sirolimus and the internal standard, respectively. The method was linear over the range 0.2 to 100.0 microg l(-1). The accuracy and inter-day precision, over this concentration range, was 94.4% to 104.4% and 1.4% to 5.0%, respectively. The accuracy and total precision at the limit of quantitation (0.2 microg l(-1)) was 103.0% and 10.8%, respectively. The mean absolute recovery of sirolimus and the internal standard were 80.5% and 81.3%, respectively. The sensitivity and analytical concentration range of the method make it suitable for therapeutic drug monitoring and pharmacokinetic studies. Further, the ability of the method to measure parent drug specifically will facilitate the evaluation of immunoassays for sirolimus.