MSACL 2023 Abstract

LC-MS/MS Analysis of Immunosuppressant Drugs in Whole Blood Using the Xevo TQ Absolute with Capitainer B Device Absorptive Microsampling for Clinical Research

Peter Harrsch(1), Stephen Balloch(2), Lisa J Calton(2), Gareth Hammond(2)
(1) Waters Corporation, 34 Maple Street, Milford, MA 01757(2) Waters Corporation, Stamford Avenue, Altrincham Road, Wilmslow, SK9 4AX, UK

Peter Harrsch, BS,PhD (Presenter) Waters Corporation >> POSTER (PDF)

Presenter Bio: Peter joined Waters Corporation in 2005 as an Executive Clinical/Forensic Specialist. Currently Peter is the Market Development Manager responsible for developing the clinical LC-MS business with the Americas sales team while interfacing with global clinical marketing. Prior to Waters, Peter held various positions as Senior Mass Spectrometry Sales Specialist at Bruker Daltonics, SFC-MS Business Development Manager at Berger Instruments, and Senior Applications Consultant for LC-MS at Agilent Technologies. Before moving to the scientific instrument segment, Peter worked in the pharmaceutical industry as a Senior Research Investigator for Sterling Pharmaceuticals and a Post-Doctoral Scientist at SmithKline focusing mainly on protein drug targets and protein-based biopharmaceuticals.
Over the years, Peter has developed numerous hyphenated mass spec methods for analysis of pharmaceuticals and drugs of abuse, protein/peptide drugs, polymers, and small molecule metabolites. Other accomplishments include the development of the first commercially available SFC-MS system. Peter has delivered many LC-MS training courses and has given poster and oral presentations at numerous scientific meetings.

Relevant Financial Disclosures (within past 24 months, reported on Mar 06, 2022) Stock/Bonds Waters Corporation Salary Waters Corporation Conflict mitigation NOT REQUIRED.

Introduction:
Traditional laboratory analysis of the immunosuppressant drugs cyclosporine, everolimus, sirolimus and tacrolimus is well-established in clinical research. However there remains a need for individuals to undergo an invasive, time-consuming and disruptive process under the supervision of trained staff in order to collect a sufficient volume of whole blood for laboratory analysis.

A reliable, remote sampling method may find utility in a clinical research setting. Here we describe the use of Capitainer™ B qDBS Devices to obtain analytically sensitive, precise and accurate data for cyclosporine, everolimus, sirolimus and tacrolimus analysis using small sample volumes.

The Waters ACQUITY™ UPLC™ I-Class System with Xevo™ TQ Absolute Mass Spectrometer was used to analyze these samples.

Methods:
An in-house laboratory developed LC-MS/MS method to analyze all four immunosuppressants in a single run was developed. Waters MassTrak™ Immunosuppressant Calibrator and Control Sets (IVD) and whole blood External Quality Assurance samples (LGC, Bury, UK) were used in conjunction with Capitainer B qDBS Devices to assess the performance of the method.

Samples (10µL) were collected using Capitainer® B qDBS devices, and the sample extracted using solvent containing internal standards. A water/methanol/ammonium acetate gradient was used with a Waters C18 HSS SB column on a Waters ACQUITY™ UPLC™ I-Class and Xevo TQ Absolute Mass Spectrometer operating in positive electrospray ionization mode with run time of less than 2 minutes.

Results:
Analytical sensitivity of the lowest calibrator at 1 ng/mL for everolimus, sirolimus and tacrolimus and 25 ng/mL for cyclosporine was demonstrated with S/N (PtP) > 10 across five analytical runs. We successfully demonstrated linearity of cyclosporine from 25-1500 ng/mL and everolimus, sirolimus and tacrolimus from 1-30 ng/mL, with r2>0.995 over five analytical runs. Total precision and repeatability across the four immunosuppressants (2, 8 and 22 ng/mL for everolimus, tacrolimus and tacrolimus; 150, 400 and 900 ng/mL for cyclosporine) with five replicates over five analytical runs (n = 25) was ≤15% CV. External quality assurance samples for all drugs met the scheme acceptance criteria, with mean bias ≤15% CV.

Conclusions:
Using Capitainer B qDBS devices and sample small volumes (10 µL) of whole blood, an in-house laboratory method was used to meet validation goals for analytical sensitivity, linearity, precision and accuracy for cyclosporine, everolimus, sirolimus and tacrolimus. Furthermore, the advantages conferred by volumetric absorptive microsampling, notably removing the requirement for travel and a venous blood draw and facilitating home sampling, render this technique applicable to clinical research.

For Research Use Only. Not for Use in Diagnostic Procedures.
ACQUITY, UPLC, Xevo, and MassTrak are trademarks of Waters Technologies Corporation. Capitainer is a trademark of Capitainer AB.