Article ID Journal Published Year Pages File Type
7617616 Journal of Chromatography B 2014 6 Pages PDF
Abstract
An ultra high-pressure liquid chromatography/mass spectrometry (UHPLC/MS) separation and analysis method has been devised for open access analysis of synthetic reactions used in the production of DNA-encoded chemical libraries. The aqueous mobile phase is 100 mM hexafluoroisopropanol and 8.6 mM triethylamine; the organic mobile phase is methanol. The UHPLC separation uses a C18 OST column (50 mm × 2.1 mm × 1.7 μm) at 60 °C, with a flow rate of 0.6 mL/min. Gradient concentration is from 10 to 40% B in 1.0 min, increasing to 95% B at 1.2 min. Cycle time was about 5 min. This method provides a detection limit of a 20-mer oligonucleotide by mass spectrometry of better than 1 pmol on-column. Linear UV response for 20-mer extends from 2 to 200 pmol/μL in concentration, same-day relative average deviations are less than 5% and bias (observed minus expected) is less than 10%. Deconvoluted mass spectra are generated for components in the predicted mass range using a maximum entropy algorithm. Mass accuracy of deconvoluted spectra is typically 20 ppm or better for isotopomers of oligonucleotides up to 7000 Da.
Related Topics
Physical Sciences and Engineering Chemistry Analytical Chemistry
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