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In many laboratories across the world, acylcarnitines and related molecules from a variety of sample types are quantified to investigate metabolic conditions, such as organic acidemias and fatty acid oxidation defects. Carnitines and acylcarnitines vary in their hydrophobicity, ranging from small, polar carnitine to acylcarnitines with very long acyl chain lengths. This variability poses a unique challenge for chromatographic method development. This assay is performed without derivatization and with minimal sample preparation to quantify these low-level analytes in a biological matrix.
SWATH Acquisition (a data independent acquisition workflow) coupled with quality spectral libraries is of increasing interest when analyzing complex biological samples due to the comprehensive and quantitative nature of the results. Here, the detection rates of metabolites in plasma from both the DDA and DIA datasets on the TripleTOF 6600 System was studied. Also a series of LC-MS/MS libraries were also evaluated in this study include NIST 17, MoNA (MassBank of North America), METLIN, and SCIEX libraries. It was found that SWATH Acquisition identifies over 70% more compounds than an optimized data dependent acquisition (DDA) method.
High sensitivity analysis is key for the analysis of lipid mediators in human plasma. Here the SCIEX lipid mediator assay has been expanded and adapted to the SCIEX 7500 System. Sensitivity was compared to the QTRAP 6500+ System and an average gain in peak areas of 53 fold was observed.
In the case of protein therapeutics, the charge heterogeneity profile needs to be characterized and monitored as it has a potential impact on a product’s safety and/or efficacy. Here, a liability study focusing on the charge heterogeneity profile via CEX in combination with the SCIEX X500B QTOF System is demonstrated. The SCIEX flexible solution for MAM within SCIEX OS Software 1.7 is employed to track the changes utilizing intact reconstructed data for each charge variant.
In this study, O. basillicum plants were grown under different light exposure conditions in a new concept microcosm to examine the underlying metabolic changes that can occur within different plant parts. SWATH acquisition, MarkerView software and library matching using the Natural Products HR-MS/MS Spectral Library provided a streamlined workflow for the discovery of differentially regulated plant metabolites.
Synthetic biology is a rapidly growing field driven by recent technology developments in gene editing and others. Large strain libraries can be developed that the must be screened for metabolomic changes. Current LC-MS/MS separation techniques are used but are often not rapid enough for efficient library screening. Here, a quantitative approach for the rapid screening of yeast strains using the Echo MS System was demonstrated, monitoring over 60 metabolites. The amount of analysis time saved was considerable, with the Echo MS system analysis time being 5.6x faster than the traditional LC-MS analysis, while still demonstrating sufficient reproducibility and sensitivity for strain profiling.
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