Determination of Tacrolimus in Plasma by Supramolecular Solvent Extraction/Ultra-high Performance Liquid Chromatography£­ Tandem Mass Spectrometry

KeyWord:tacrolimus  immunosuppresive agent  supramolecular solvent  plasma  ultra-high performance liquid chromatography£­tandem mass spectrometry

Abstract:

      An analytical method for the determination of tacrolimus£¨TAC£© in blood plasma was established by supramolecular solvent£¨SUPRAS£© extraction combined with ultra-high performance liquid chromatography£­tandem mass spectrometry£®After optimizing the key factors such as the composition and amount of SUPRAS£¬and vortex extraction time through single factor experiment and response surface design£¬blood plasma samples were extracted efficiently with SUPRAS formed by pentanol£¬tetrahydrofuran and water£®The extract was separated on a Waters ACQUITY UPLC BEH C18 column£¨50 mm ¡Á 2.1 mm£¬1.7 ¦Ìm£©£¬analyzed by electrospray ionization mass spectrometry in positive ion mode under multiple reaction monitoring£¨MRM£© mode£¬and quantified by internal standard method£®Experimental results demonstrated that there was a good linear relationship for TAC in the concentration range of 0.5£­30 ng/mL£¬with a correlation coefficient£¨r£© of 0.998 6£®The limit of detection£¨LOD£© and quantitation£¨LOQ£© were 0.1 ng/mL and 0.5 ng/mL£¬respectively£®The average recoveries£¨n = 3£© at low£¬medium and high spiked concentration levels ranged from 91.9% to 99.9%£¬with relative standard deviations£¨RSDs£© of 1.7%£­5.7%£®The proposed method is rapid£¬sensitive and stable£¬and it was suitable for the accurate determination of TAC in blood plasma.