Sensitive, wide‐range and high‐throughput quantification of cyclosporine in whole blood using ultra‐performance liquid chromatography coupled to tandem mass spectrometry and comparison with an antibody‐conjugated magnetic immunoassay

Abstract

Because either trough or peak concentration at 2 h after administration is measured in routine therapeutic drug monitoring for cyclosporine A (CyA), a quantification method with a wide-range calibration curve capable of simultaneously measuring both concentrations is required. We developed a sensitive, wide-range and highthroughput quantification method for CyA in whole blood using ultra-performance liquid chromatography-tandem mass spectrometry (UPLC-MS/MS), and compared patients' blood CyA levels measured by UPLC-MS/MS and antibody-conjugated magnetic immunoassay (ACMIA). Whole blood samples were prepared by solid-phase extraction using Oasis HLB μElution plate. The UPLC-MS/MS assay showed excellent linearity over a wide calibration range of 5-2500 ng/mL. Within-batch accuracy and precision as well as batch-to-batch accuracy and precision fulfilled the criteria of US Food and Drug Administration guidelines. The blood CyA concentrations measured by the UPLC-MS/MS assay correlated strongly with those measured by ACMIA. A Bland-Altman plot showed a fixed error between CyA concentrations measured by the two methods, and the concentrations measured by the UPLC-MS/MS method were consistently lower than those measured by ACMIA. We have succeeded to develop a sensitive, wide-range and high-throughput quantification method for CyA in whole blood using UPLC-MS/MS. K E Y W O R D Santibody-conjugated magnetic immunoassay, cyclosporine, therapeutic drug monitoring, ultraperformance liquid chromatography-tandem mass spectrometry | INTRODUCTIONCyclosporine A (CyA) is a calcineurin inhibitor and exhibits immunosuppressive effect by inhibiting cytokines, particular interleukin-2, which are secreted by T lymphocytes (Taylor et al., 2005). Like tacrolimus, CyA is an important immunosuppressant used in various organ transplantations and autoimmune diseases (Kaufman et al., 2004). CyA has strong pharmacological activity at low doses, but its therapeutic window is narrow. Furthermore, due to the large intra-and inter-individual variability in pharmacokinetics as well as significant disposition in erythrocytes,

Positive correlation between organic anion transporter 1B function indicated by plasma concentration of coproporphyrin‐I and blood concentration of cyclosporin A in real‐world patients

“…CyA concentration was measured according to the method described in our previous report 22 . Briefly, 50 μL of whole blood was pretreated by a solid‐phase extraction method using Oasis HLB μElution Plate.…”

“…CyA concentration was measured according to the method described in our previous report. 22 Briefly, 50 μL of whole blood was pretreated by a solid-phase extraction method using Oasis HLB μElution Plate. The extracted samples were analysed by ultrahigh-performance LC coupled to tandem MS (UHPLC-MS/MS; Xevo TQD; Waters, Milford, MA, USA).…”

Metabolomics for personalized medicine: the input of analytical chemistry from biomarker discovery to point-of-care tests

“…Most of them deal with drugs and their metabolites, and, to a lesser extent, with endogenous metabolites such as steroids, thyroid hormones, and vitamin D derivatives. Some of these studies show that these two methods exhibit the same kind of performances [158][159][160], whereas others point out variable quantitative performances between immunoassays [161], and specificity issue with over-or underestimated concentrations measured in immunoassays [162][163][164][165].…”

Pharmacokinetics and bioequivalence of two cyclosporine oral solution formulations in cats

“…It is a powerful immunosuppressant drug that acts by inhibiting the proliferation of T-lymphocytes ( 6 , 7 ). CyA's direct effect is via inhibition of calcineurin and exhibits an immunosuppressive effect by inhibiting cytokines, which are secreted by T lymphocytes ( 8 – 10 ). Cyclosporine is lipophilic, distributes widely, and is stored in the skin and adipose tissue.…”