Mary E. Lame, Erin E. Chambers, and Kenneth J. Fountain
Waters Corporation, Milford, MA, USA
Application Advantages â– The sample volume is reduced by 2 times and the sensitivity is increased by 10 times. It is convenient to carry out multiple injections of samples and better meet the needs of ISR repeat analysis.
â– Reduce solvent consumption by 50 times and save on analysis costs.
â– Mixed mode SPE reduces matrix interference and enhances the selectivity of bradykinin extraction in plasma.
■The Oasis μElution 96-well plate concentrates the sample and minimizes peptide loss, allowing the detection limit of bradykinin in plasma to reach 5 pg/mL.
â– Highly selective rapid SPE extraction (<30 minutes) eliminates the time-consuming immunoaffinity purification step.
Waters Solutions
ionKey/MS® system
ACQUITY UPLC® M-Class
IonKeyTM source
Xevo® TQ-S
iKeyTM separation unit
MassLynxTM
Oasis® WCX 96-well μElution board
ACQUITY® acquisition board
Introduction <br> of peptide material stable, demand is sensitive analysis of common challenges of chromatographic separation and mass spectrometry. Common peptides are often difficult to analyze by LC-MS/MS because they are less susceptible to ionization and are less prone to produce ideal fragment ions, resulting in lower MS sensitivity, making LC and sample preparation methods very difficult to develop. The previous application note (720004833ZH) details how to develop a fast and flexible SPE-LC-MS/MS analysis method for the quantification of bradykinin (Figure 1) in human plasma; bradykinin is preclinical or Important biomarkers in the drug development phase 1 . Since the concentration of bradykinin in plasma is as low as pg/mL and the metabolism is fast, it can be artificially produced by proteolysis during blood collection and sample preparation, so accurate quantification is quite difficult.
In this experiment, the entire LC-MS platform was updated using the ionKey/MS system, which integrates UPLC® analysis into the mass spectrometer source (Figure 2). As shown in Figure 3, the iKey separation unit (150 μm id) contains flow paths, electronics, ESI interface, heater, eCord, and chromatographic packing to achieve UPLC separation performance. In addition, this technique significantly improves sensitivity compared to 2.1 mm id chromatography and is an ideal method for peptide analysis. Most bioanalytical LC-MS/MS methods often consume large amounts of solvent and sample, increasing the cost of analysis and limiting the number of repeatable analyses. In addition to increased sensitivity, ionKey/MS reduces solvent and sample consumption, providing enough samples for repeated injections to better meet ISR testing needs.
To download the application minutes, please click: http://?lid=134786602&cid=511436
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