ACQUITY™ UPLC™ I-Class SM-FL/Xevo™ TQ Absolute System: Analytical Performance for Free Testosterone
Este es un resumen de la aplicación y no contiene una sección experimental detallada.
Abstract
The Waters™ ACQUITY UPLC I-Class SM-FL/Xevo TQ Absolute 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 SM-FL/Xevo TQ Absolute IVD System for the analysis of free testosterone in serum.
Introduction
The Waters™ ACQUITY UPLC I-Class SM-FL/Xevo TQ Absolute 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 SM-FL/Xevo TQ Absolute IVD System for the analysis of free testosterone in serum.
Experimental
The ACQUITY UPLC I-Class SM-FL/Xevo TQ Absolute IVD System was controlled by MassLynx™ Software (v4.2) and the data processed using the TargetLynx™ XS Application Manager. Calibrators were prepared in a 52.75 mM HEPES buffer adjusted to pH 7.4, to match the constitution of the diasylate obtained during sample preparation. Samples were processed using the following conditions:
Sample Preparation Conditions
200 µL sample was processed using an equilibrium dialysis incubation, then treated with methyl tert-butyl ether, and a liquid-liquid extraction procedure was performed to obtain the final extract for analysis.
LC Conditions
|
Sample manager: |
SM-FL (PL, Load Ahead disabled) |
|
Solvent manager: |
BSM |
|
Column: |
ACQUITY UPLC BEH™ C18 1.7 µm, 2.1 mm x 100 mm |
|
Mobile phase A: |
0.2 mM Ammonium fluoride in water |
|
Mobile phase B: |
0.2 mM Ammonium fluoride in methanol |
|
Flow rate: |
0.30 mL/min |
|
Gradient: |
50% B for 0.5 minutes, 55% B at 0.5 minutes, 55–75% B over 2.7 minutes, 75–98% B over 0.05 minutes, hold 98% B for 0.45 minutes, equilibrate with 50% B for 0.7 minutes |
MS Conditions
|
Resolution: |
MS1 (0.75 FWHM), MS2 (0.75FWHM) |
|
Acquisition mode: |
MRM |
|
Polarity: |
ESI (+) |
Results and Discussion
Performance characteristics of free testosterone on the ACQUITY UPLC I-Class SM-FL/Xevo TQ Absolute IVD System are shown in Table 1. Analytical sensitivity of the chromatographic separation is illustrated in Figure 1.
Table 1. Performance characteristics of free testosterone. Range defined by linear fit where r2 >0.995. LLOQ defined by S/N (PtP) >10 and %RSD ≤20%. %RSD at LLOQ determined through analytical sensitivity experiments performed over five occasions (n=50). Total precision and repeatability of QCs performed over five occasions in serum (n=25). EQA mean bias determined by comparison of obtained values to the all laboratory trimmed mean (ALTM) value (n=45).
Figure 1. Chromatogram showing the analysis of free testosterone using the ACQUITY UPLC I-Class SM-FL/ Xevo TQ Absolute IVD System.
Conclusion
The Waters ACQUITY UPLC I-Class SM-FL/Xevo TQ Absolute IVD System has demonstrated the capability to analyse free testosterone in serum samples whilst delivering analytical sensitivity and precision.
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.
720008233, February 2024
