• Application Note

ACQUITY UPLC I-Class/Xevo TQD IVD System: Analytical Performance for an Antifolate Agent

ACQUITY UPLC I-Class/Xevo TQD IVD System: Analytical Performance for an Antifolate Agent

  • Waters Corporation

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.

ACQUITY UPLC I-Class/Xevo TQD IVD System
ACQUITY UPLC I-Class/Xevo TQD IVD System.

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.

Performance characteristics of methotrexate
Table 1. Performance characteristics of methotrexate. Range defined by linear fit where r2 >0.99. LLOQ defined by S/N (PtP) >10 and %RSD ≤20%. %RSD at LLOQ determined through analytical sensitivity experiments performed over three occasions (n=30). Total precision and repeatability of QCs performed over five occasions in plasma (n=25). EQA mean bias determined by comparison of obtained values to the gravimetric reference value.
Note: To convert SI units to conventional mass units divide by 2.2005 (μmol/L to μg/mL).
Chromatogram showing the analysis of methotrexate using the ACQUITY UPLC I-Class/Xevo TQD IVD System
Figure 1. Chromatogram showing the analysis of methotrexate using the ACQUITY UPLC I-Class/Xevo TQD IVD System.

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

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