Simultaneous quantification of cyclosporin, tacrolimus, sirolimus and everolimus in whole blood by UHPLC–MS/MS for therapeutic drug monitoring

Abstract

The aim of this study was to develop and validate a UHPLC–MS/MS assay to quantify cyclosporin (CYC), tacrolimus (TAC), sirolimus (SIR) and everolimus (EVE) in human whole blood for therapeutic drug monitoring. Analytes were extracted from 50 μL human whole blood by protein precipitation. The separation of the drugs was performed on an Acquity UPLC BEH C18 column. Analytes were eluted with a mobile phase consisting of 2 mM ammonium acetate with 0.1% formic acid (v/v) in deionised water and 2 mM ammonium acetate with 0.1% formic acid (v/v) in methanol at a flow rate of 300 μL/min in gradient elution. The method performance was evaluated by analysing patient blood samples and/or external quality control samples [proficiency testing (PT) scheme]. The method was linear from 23.75 to 1094.0, 1.3 to 42.4, 1.3 to 47.0 and 1.2–41.6 μg/mL for CYC, TAC, SIR and EVE, respectively. The within‐ and between‐assay reproducibility results were ˂ 11%. Results from PT and patient sample quantification were comparable to those obtained previously by an in‐house validated method using protein precipitation and liquid–liquid extraction. This method showed good analytical performance for quantifying CYC, TAC, SIR and EVE in whole blood over their respective calibration ranges.

High-Sensitivity and High-Throughput Quantification of Everolimus in Human Whole Blood Using Ultrahigh-Performance Liquid Chromatography Coupled With Tandem Mass Spectrometry

“…Numerous sole or simultaneous methods for measuring blood EVR concentration using HPLC-MS/MS or UHPLC-MS/MS have been developed to date. [9][10][11][12][13][18][19][20][21][22][23][24][25][26][27][28][29][30][31] To the best of our knowledge, the method developed by Rickert et al 21 has the widest calibration curve range, but the quotient of the upper and lower limits is 400, which is narrower than that of our method (500). Furthermore, among the methods reported, those of Rickert et al 21 and Taibon et al 22 have the lowest limit of calibration range of 0.25 ng/mL, which is higher than that of our method (0.1 ng/mL).…”

“…Most of the previous reports on HPLC-MS/MS or UHPLC-MS/MS methods for blood EVR concentration measurement used deproteinization as the sample pretreatment method. 10,12,13,[18][19][20]23,24,[28][29][30] This pretreatment method removes proteins by adding an organic solvent, such as acetonitrile, to whole blood followed by centrifugation. This method is relatively easy, but matrix removal is inadequate.…”

“…8 The latter include high-performance liquid chromatography coupled with tandem mass spectrometry (HPLC-MS/MS) 10,11 and ultrahigh-performance liquid chromatography coupled with tandem mass spectrometry (UHPLC-MS/ MS). 12,13 LTIA is generally used in clinical practice because of its ease of operation and short measurement time; however, this method has several limitations. The lower limit of quantification (LLOQ) of 2.0 ng/mL is high and close to the lower limit of the target trough concentration range.…”

Non-immobilized GO-SELEX screening of aptamers against cyclosporine A and its application in a AuNPs colorimetric aptasensor

“…10,13,14 There are various methods reported for monitoring CsA, mainly including chromatography and immunoassay techniques. [15][16][17][18][19][20][21] Although the immunoassay is relatively sensitive, it has some problems, such as poor repeatability, cross-reaction, and high cost. Compared with the immunoassay, chromatography shows better selectivity, accuracy, and lower variability.…”

Development of UHPLC–MS/MS method for simultaneous determination of tacrolimus and sirolimus in human whole blood and comparisons with two immunoassays

“…The quantification and comparison of immunosuppressive drugs using LC-MS/MS and immunoassay have already been reported (Antunes et al, 2021;Bodnar-Broniarczyk et al, 2019;Koster et al, 2009;Koster et al, 2013;Mei et al, 2018;Schmid et al, 2009;Shigematsu et al, 2020). However, few studies compare chemiluminescence microparticle immunoassay (CMIA), electrochemiluminescence immunoassay (ECLIA) and UHPLC-MS/MS at the same time.…”