Mass Spectrometry Quantitation of Immunosuppressive Drugs in Clinical Specimens Using Online Solid-phase Extraction and Accurate-mass Full Scan-single Ion Monitoring

Overview

Journal J Mass Spectrom Adv Clin Lab

Specialty General Medicine

Date 2023 Mar 20

PMID 36937810

Authors

Priscilla S-W Yeung

Paige Miller

Tran Bao Lai-Nyugen

Phil Cheng

Amira Ibrahim

Run-Zhang Shi

Raffick A R Bowen

Ruben Yiqi Luo

Affiliations

Soon will be listed here.

Abstract

Introduction: Therapeutic drug monitoring (TDM) of immunosuppressants is essential for optimal care of transplant patients. Immunoassays and liquid chromatography-mass spectrometry (LC-MS) are the most commonly used methods for TDM. However, immunoassays can suffer from interference from heterophile antibodies and structurally similar drugs and metabolites. Additionally, nominal-mass LC-MS assays can be difficult to optimize and are limited in the number of detectable compounds.

Objectives: The aim of this study was to implement a mass spectrometry-based test for immunosuppressant TDM using online solid-phase extraction (SPE) and accurate-mass full scan-single ion monitoring (FS-SIM) data acquisition mode.

Methods: LC-MS analysis was performed on a TLX-2 multi-channel HPLC with a Q-Exactive Plus mass spectrometer. TurboFlow online SPE was used for sample clean up. The accurate-mass MS was set to positive electrospray ionization mode with FS-SIM for quantitation of tacrolimus, sirolimus, everolimus, and cyclosporine A. MS fragmentation pattern was used for compound confirmation.

Results: The method was validated in terms of precision, analytical bias, limit of quantitation, linearity, carryover, sample stability, and interference. Quantitation of tacrolimus, sirolimus, everolimus, and cyclosporine A correlated well with results from an independent reference laboratory (r = 0.926-0.984).

Conclusions: Accurate-mass FS-SIM can be successfully utilized for immunosuppressant TDM with good correlation with results generated by standard methods. TurboFlow online SPE allows for a simple "protein crash and shoot" sample preparation protocol. Compared to traditional MRM, analyte quantitation by FS-SIM facilitates a streamlined assay optimization process.

Citing Articles

High-resolution mass spectrometry measurement of N-terminal carbamylated hemoglobin as a potential marker for chronic diseases with elevated blood urea levels.

Chen F, Yeung P, Wong C, Luo R J Mass Spectrom Adv Clin Lab. 2025; 35:8-13.

PMID: 40034798 PMC: 11873320. DOI: 10.1016/j.jmsacl.2025.01.001.

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