title: Reaction profiling by ultra high-pressure liquid chromatography/time-of-flight mass spectrometry in support of the synthesis of DNA-encoded libraries.
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 100mM hexafluoroisopropanol and 8.6mM triethylamine; the organic mobile phase is methanol. The UHPLC separation uses a C18 OST column (50mm×2.1mm×1.7μm) at 60°C, with a flow rate of 0.6mL/min. Gradient concentration is from 10 to 40% B in 1.0min, increasing to 95% B at 1.2min. Cycle time was about 5min. This method provides a detection limit of a 20-mer oligonucleotide by mass spectrometry of better than 1pmol on-column. Linear UV response for 20-mer extends from 2 to 200pmol/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 20ppm or better for isotopomers of oligonucleotides up to 7000Da.
authors.affiliation: Lexicon Pharmaceuticals, 350 Carter Road, Princeton, NJ 08540 USA. Electronic address: lhargiss@verizon.net.
authors.forename: Leonard O
authors.initials: LO
authors.lastname: Hargiss
authors.affiliation: Lexicon Pharmaceuticals, 350 Carter Road, Princeton, NJ 08540 USA.
authors.forename: G Greg
authors.initials: GG
authors.lastname: Zipp
authors.affiliation: Lilly Centro de Investigación, Avda. de la Industria, 30, 28108 Alcobendas, Madrid, Spain.
authors.forename: Theodore C
authors.initials: TC
authors.lastname: Jessop
authors.affiliation: Lexicon Pharmaceuticals, 350 Carter Road, Princeton, NJ 08540 USA.
authors.forename: Xuejun
authors.initials: X
authors.lastname: Sun
authors.affiliation: Lexicon Pharmaceuticals, 350 Carter Road, Princeton, NJ 08540 USA.
authors.forename: Philip
authors.initials: P
authors.lastname: Keyes
authors.affiliation: Lexicon Pharmaceuticals, 350 Carter Road, Princeton, NJ 08540 USA.
authors.forename: David B
authors.initials: DB
authors.lastname: Rawlins
authors.affiliation: Lexicon Pharmaceuticals, 350 Carter Road, Princeton, NJ 08540 USA.
authors.forename: Zhi
authors.initials: Z
authors.lastname: Liang
authors.affiliation: Lexicon Pharmaceuticals, 350 Carter Road, Princeton, NJ 08540 USA.
authors.forename: Kum Joo
authors.initials: KJ
authors.lastname: Park
authors.affiliation: Lexicon Pharmaceuticals, 350 Carter Road, Princeton, NJ 08540 USA.
authors.forename: Huizhong
authors.initials: H
authors.lastname: Gu
mesh_terms: D000465:Algorithms; D002851:Chromatography, High Pressure Liquid / Q000379:methods*; D004247:DNA / Q000138:chemical synthesis / Q000737:chemistry*; D015723:Gene Library; D013058:Mass Spectrometry / Q000379:methods*; D009841:Oligonucleotides / Q000032:analysis / Q000737:chemistry*; D021241:Spectrometry, Mass, Electrospray Ionization; D013816:Thermodynamics
chemical_list: D009841:Oligonucleotides; D004247:DNA
keywords: DNA-encoded libraries; Electrospray ionization; Mass spectrometry; Maximum entropy; Oligonucleotides; UHPLC