Abstract
Quantitative nontarget analysis is intended to provide a measurement of concentration of newly identified components in complex biological or environmental samples for which authentic or labeled standard do not exist. Electrospray ionization-high field asymmetric waveform ion mobility spectrometry-mass spectrometry (ESI-FAIMS-MS) has unique advantages that allowed us to develop a novel approach for quantification of nontarget analytes. In the nontarget analysis of urinary metabolites by ESI-FAIMS-MS, we find it practical and beneficial to analyze highly diluted urine samples. We show that urine extracts can be analyzed directly at very high dilutions (up to 20,000 times) by extending MS analysis times during slow FAIMS scanning. We explore the effects of sample dilution on ionization efficiency and ionization suppression in direct electrospray of complex sample matrixes. We consistently observe two distinct regimes in ESI operation related to the limited ionization capacity of this method. In the linear dynamic concentration range below the limiting ionization capacity, the analytical sensitivity of an analyte is constant and does not depend on matrix composition and concentration. Once the capacity of ESI is exceeded, all species exhibit log-log linearity in signal response. We show how quantification can be carried out using two different approaches, one for analytes which can be detected in the linear regime and another for those only detected in the suppression regime that overcomes the effects of ionization suppression. Our new insight into ionization suppression effects in ESI is of broad interest to anyone using ESI as an ionization technique for the MS analysis of complex samples.
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