A Rapid Analytical Platform for Biofluid Profiling In Discovery Metabolomics and Lipidomics

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Adam King, Lee Gethings, Robert Plumb, Lauren Mullin, Ian Wilson, Garth Maker, Robert Trengove
Waters, Imperial College, Murdoch University
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Metabolic phenotyping has been widely employed in large epidemiological studies in the effort to discovery more about health and disease. UPLC and hybrid mass spectrometers have been essential tools in analyzing the matrices chosen for these studies. However, long acquisition times per samples (>15 mins) has meant large sets, typically employed in epidemiological and biobanking studies, can take days to analyze, ultimately putting strain on resources. Furthermore, this has led to sample acquisition being performed across multiple runs for the same data set, risking the development of batch effects when attempting to recombine the data. In order to address these issues, a suite of rapid profiling methods have been developed, reducing run time and solvent consumption by 75%. These methods were employed for the analysis of rodent urine and human plasma lipid extracts from breast cancer patients, using a Waters Acquity I-class UPLC system coupled to a Synapt G2-Si QTof mass spectrometer. Standard LC-MS methods for metabolite and lipid profiling using 2.1 mm i.d. columns were geometrically scaled, reducing the i.d., analysis times, injection volumes and mobile phase flow rate, while ultimately increasing the linear velocity. Each scaling demonstrated a preservation of the retention mechanism with relative retention times of probe compounds being maintained. Incorporating the ion mobility schema improved resolution of coeluting ions, ultimately improving spectral clarity and with the generation of collisional cross section values, increased confidence in identification through database searches.

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