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Liquid Chromatography, Multiple Reaction Monitoring by Mass Spectrometry (LC-MRM-MS) method, using a stable isotope-labeled internal standard has shown major progress in biological analysis. In this approach, optimizing the selected precursor ion in Q1 and product ion in Q3, for each monitoring transition, is important for establishing the most sensitive MRM assay. The sensitivity and accuracy improvement requires an experimental study for determination of optimal instrumental parameters, in order to generate abundant quantities of precursor and product ion pair. The use of LC allows a fast separation, generating narrow peaks, reproducible retention time and peak area. The control of chromatographic parameter is performed by system suitability test. The use of Scheduled MRM requires determination of dwell and MRM detection window. The dwell time is the time spent for recording the signal intensity of one data point; a longer dwell time results in more accurate measurements of signal intensity. All these instrument settings are important to determine the exact amount of scans time of a particular event, leading to the best quality and quantity data possible.

Evaluation of Scheduled MRM parameters: a powerful tool for analysis of modified carrier protein of meningococcal polysaccharide C conjugate vaccine