A rapid HPLC-MS/MS method for the simultaneous quantification of cyclosporine A, tacrolimus, sirolimus and everolimus in human blood samples

Abstract

In the past few years, high-performance liquid chromatography tandem mass spectrometry (HPLC-MS/MS) has matured to a true alternative to antibody-based immunoassays in routine therapeutic drug monitoring. In transplantation medicine, mass spectrometry-based assessment of immunosuppressant drug levels is considered a gold standard diagnostic procedure. We describe a fast state-of-the-art routine online solid-phase extraction (SPE) HPLC-MS/MS analysis platform that allows monitoring of cyclosporine A, tacrolimus, sirolimus and everolimus from 50-microl aliquots of EDTA whole blood specimens within 3.4 min total analysis time. Sample purification is done by offline protein precipitation followed by two automated chromatographic separation steps. Mass spectrometry-based analyte quantification relies on selected reaction monitoring experiments. The assay underwent complete validation and performance evaluation studies and performs very well in several international proficiency testing schemes. In daily routine, it allows reporting of about 75 patient sample results per work shift with a typical total individual sample turnaround time of less than 3 h.

Liposomes containing bile salts as novel ocular delivery systems for tacrolimus (FK506): in vitro characterization and improved corneal permeation

“…23 Chromatographic separation was performed on an Agilent 1200 system composing of a quaternary pump, a degasser, an autosampler, and a column heater (Agilent Technologies Inc). The high-performance liquid chromatography column was a Zorbax XDB-C18 (2.1 × 50 mm, 5 µm) column from Agilent.…”

A comparison of liquid chromatography-tandem mass spectrometry (LC-MS/MS) and enzyme-multiplied immunoassay technique (EMIT) for the determination of the cyclosporin A concentration in whole blood from Chinese patients

“…This technique offers analytical specificity superior to that of immunoassays or conventional highperformance/pressure liquid chromatography (HPLC) for low molecular weight analytes. LC-MS/MS has superior analytical specificity and lower reagent cost, especially if one considers the ability to multiplex several different immunosuppressive drugs (8)(9)(10). LC-MS/MS is a sensitive and selective technique that could prove useful in TDM for CyA.…”

Plasma clot formation and clot lysis to compare effects of different anticoagulation treatments on hemostasis in patients with atrial fibrillation

“…DOAC quantification by LC-MS/MS was facilitated by utilizing a previously established two-dimensional chromatography setup (Agilent Technologies, Waldbronn, Germany) hyphenated to an API 4000 Qtrap instrument (ABSciex, Framingham, MA, USA) operated in the electrospray ionization (ESI) mode [11]. Based on this instrumentation, a research-use-only assay was used as previously published [2].…”