Dynamic Data-Independent Acquisition Mass Spectrometry with Real-Time Retrospective Alignment

Data-independentacquisition (DIA) mass spectrometry has grownin popularity in recent years, because of the reproducibility andquantitative rigor of a systematic tandem mass spectrometry (MS/MS)sampling method. However, traditional DIA methods may spend valuableinstrument time acquiring MS/MS spectra with no usable informationin them, affecting sensitivity and quantitative performance. We developeda DIA strategy that dynamically adjusts the MS/MS windows during thechromatographic separation. The method focuses MS/MS acquisition onthe most relevant mass range at each point in time Rincreasingthe quantitative sensitivity by increasing the time spent on eachDIA window. We demonstrate an improved lower limit of quantification,on average, without sacrificing the number of peptides detected.