ACQUITY UPLC I-Class/Xevo TQD IVD System: Analytical Performance for an Antifolate Agent
For in vitro diagnostic use. Not available in all countries.
Introduction
The Waters ACQUITY UPLC I-Class/Xevo TQD IVD System enables the quantification of organic compounds in human biological liquid matrices.
This document describes a test of the analytical performance of the ACQUITY UPLC I-Class/Xevo TQD IVD System for the analysis of methotrexate in plasma.

Experimental
The ACQUITY UPLC I-Class/Xevo TQD IVD System was controlled by MassLynx IVD Software (v4.1) and the data processed using the TargetLynx Application Manager. Calibrators and Quality Controls were prepared by spiking commercially available reference material in plasma. The samples were processed using the following conditions:
Sample Preparation Conditions
50 μL sample was processed with methanol and centrifuged, then subsequently diluted with water prior to analysis.
LC Conditions
Column: |
ACQUITY UPLC HSS C18 SB 1.8 μm, 2.1 mm × 30 mm |
Mobile phase A: |
2 mM Ammonium acetate + 0.1% formic acid in water |
Mobile phase B: |
2 mM Ammonium acetate + 0.1% formic acid in methanol |
Flow rate: |
0.4 mL/min |
Gradient: |
77% A isocratic for 5 min |
MS Conditions
Resolution: |
MS1 (0.75 FWHM), MS2 (0.75 FWHM) |
Acquisition mode: |
MRM |
Polarity: |
ESI (+) |
Results and Discussion
Performance characteristics of methotrexate on the ACQUITY UPLC I-Class/Xevo TQD IVD System are shown in Table 1. Analytical sensitivity of the chromatographic separation is illustrated in Figure 1.

Note: To convert SI units to conventional mass units divide by 2.2005 (μmol/L to μg/mL).

Conclusion
The Waters ACQUITY UPLC I-Class/Xevo TQD IVD System has demonstrated the capability to deliver an analytically sensitive and precise method for methotrexate in plasma.
Disclaimer
The analytical performance data presented here is for illustrative purposes only. Waters does not recommend or suggest analysis of the analytes described herein. These data are intended solely to demonstrate the performance capabilities of the system for analytes representative of those commonly analyzed using liquid chromatography and tandem mass spectrometry. Performance in an individual laboratory may differ due to a number of factors, including laboratory methods, materials used, intra-operator technique, and system conditions. This document does not constitute a warranty of merchantability or fitness for any particular purpose, express or implied, including for the testing of the analytes in this analysis.
720006318, August 2018