Abstract
We describe a multiple reaction monitoring positive ion HPLC/tandem mass spectrometric method for quantification of tacrolimus in human whole blood with online extraction and cleanup. Included in this procedure: API 2000 triple quadrupole mass spectrometer with turbo-ion spray source (Applied Biosystems, Foster City, CA); 10-port diverter/switching valve (Valco, Houston, TX); HPLC system (Agilent Technologies series 1100, Wilmington, DE); 10 mm (C18) guard cartridge (Perkin Elmer, Norwalk, CT) used as an extraction column; a Nova-Pak C18 analytical column (2.1 × 150 mm I.D., 4 μm, Waters Corp, Milford, MA); washing solution, methanol: 30 mM ammonium acetate pH 5.1 (80:20); eluting solution, methanol:30 mM ammonium acetate pH 5.1 (97:3); flow rate 0.8 mL/min; and a run-time of 2.8 min. The first and third quadrupoles were set to detect the ammonium adduct ion and a high mass fragment of tacrolimus (m/z 821.5→768.3), and of an internal standard (ascomycin) (m/z 901.8→834.4). The lower limit of quantification of this method is 3.75 mg/L. The concentration of drug is determined by comparing peak-area ratios for tacrolimus and internal standard to a standard curve constructed using non-weighted linear through zero regression.
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© 2010 Humana Press, a part of Springer Science+Business Media, LLC
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Donaldson, K.J., Shaw, L.M. (2010). Quantitation of Tacrolimus in Whole Blood Using High Performance Liquid Chromatography-Tandem Mass Spectrometry (HPLC-MS-MS). In: Garg, U., Hammett-Stabler, C. (eds) Clinical Applications of Mass Spectrometry. Methods in Molecular Biology, vol 603. Humana Press. https://doi.org/10.1007/978-1-60761-459-3_47
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DOI: https://doi.org/10.1007/978-1-60761-459-3_47
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