Simultaneous Determination of Cyclosporine A, Tacrolimus, Sirolimus, and Everolimus in Whole-Blood Samples by LC-MS/MS

Abstract

<jats:p><jats:italic>Objectives</jats:italic>. Cyclosporine A (CyA), tacrolimus (TRL), sirolimus (SIR), and everolimus (RAD) are immunosuppressive drugs frequently used in organ transplantation. Our aim was to confirm a robust sensitive and selective liquid chromatography-tandem mass spectrometry (LC-MS/MS) method for determination of CyA, TRL, SIR, and RAD in whole-blood samples.<jats:italic>Materials and Methods</jats:italic>. We used an integrated online solid-phase extraction-LC-MS/MS system and atmospheric pressure ionization tandem mass spectrometry (API-MS/MS) in the multiple reaction monitoring (MRM) detection mode. CyA, TRL, SIR, and RAD were simultaneously analyzed in whole blood treated with precipitation reagent taken from transplant patients.<jats:italic>Results</jats:italic>. System performance parameters were suitable for using this method as a high-throughput technique in clinical practice. The high concentration of one analyte in the sample did not affect the concentration of other analytes. Total analytical time was 2.5 min, and retention times of all analytes were shorter than 2 minutes.<jats:italic>Conclusion</jats:italic>. This LC-MS/MS method can be preferable for therapeutic drug monitoring of these immunosuppressive drugs (CyA, TRL, SRL, and RAD) in whole blood. Sample preparation was too short and simple in this method, and it permits robust, rapid, sensitive, selective, and simultaneous determination of these drugs.</jats:p>

Ultra high performance liquid chromatography–tandem mass spectrometry method for cyclosporine a quantification in biological samples and lipid nanosystems

“…This was above 98.72% from whole blood samples, values comparable to those reported by other groups [22,31]. Recoveries from homogenate tissue samples were about 85.12±7.71% (table 3).…”

An UPLC–MS/MS method coupled with automated on-line SPE for quantification of tacrolimus in peripheral blood mononuclear cells

“…Capron et al highlighted that TAC concentrations in peripheral mononuclear blood cells (PBMCs) significantly correlate with both the development and the severity of rejection: lower intra-PCMCs TAC levels were significantly associated to histological rejection and, unlike whole-blood levels, these levels were significantly correlated to the onset of rejection episodes 1 week after transplantation [2]. Several HPLC, LC-MS and LC-MS/MS methods were reported for TAC quantification, but most of these evaluated TAC in whole blood [11][12][13][14]. Capron et al published a method to measure TAC concentrations in PBMCs: quantification of TAC was performed by LC-MS/MS after a cumbersome and time-expensive extraction procedure [15].…”

Label-free quantification of Tacrolimus in biological samples by atomic force microscopy

“…Several assays have been developed for the quantification of the drug in biological samples. The most common methods for routine FK506 monitoring are monoclonal antibody-based immunoassays, while liquid chromatography-tandem mass spectrometry (LC-MS) is currently considered as the reference method [4][5][6][7]. LC-MS provides the most specific routine drug measurement, whereas immunoassays show a notable crossreactivity with inactive metabolites [4,8], especially MIII and MV [9].…”