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  1. Artikel ; Online: Mobile Phase Aging and Its Impact on Electrospray Ionization of Oligonucleotides.

    Guimaraes, Guilherme J / Saad, Jack G / Annavarapu, Vidya / Bartlett, Michael G

    Journal of the American Society for Mass Spectrometry

    2023  Band 34, Heft 12, Seite(n) 2691–2699

    Abstract: The implementation of fluoroalcohol/alkylamine mobile phase systems in oligonucleotide LC-MS provides a good balance between chromatographic separations and MS sensitivity. Since its introduction, several parameters including mobile phase composition, ... ...

    Abstract The implementation of fluoroalcohol/alkylamine mobile phase systems in oligonucleotide LC-MS provides a good balance between chromatographic separations and MS sensitivity. Since its introduction, several parameters including mobile phase composition, additive concentration, alkylamine hydrophobicity, and different fluoroalcohols have been carefully evaluated and optimized. While our understanding of this mobile phase system has increased over the years, there are challenges that continue to hinder method performance and remain poorly understood. One of these challenges is the constant loss of MS sensitivity over time, commonly termed mobile phase aging. This study investigates two aging mechanisms associated with loss of MS sensitivity: alkylamine oxidation and aggregate formation. The relationship between pH, organic solvent, oxygen, and mobile phase aging is characterized, and mitigation strategies to extend mobile phase lifetime are discussed.
    Mesh-Begriff(e) Oligonucleotides/chemistry ; Spectrometry, Mass, Electrospray Ionization/methods ; Chromatography, Liquid/methods ; Indicators and Reagents ; Solvents
    Chemische Substanzen Oligonucleotides ; Indicators and Reagents ; Solvents
    Sprache Englisch
    Erscheinungsdatum 2023-11-18
    Erscheinungsland United States
    Dokumenttyp Journal Article
    ZDB-ID 1073671-2
    ISSN 1879-1123 ; 1044-0305
    ISSN (online) 1879-1123
    ISSN 1044-0305
    DOI 10.1021/jasms.3c00264
    Datenquelle MEDical Literature Analysis and Retrieval System OnLINE

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  2. Artikel ; Online: Alkylamine ion-pairing reagents and the chromatographic separation of oligonucleotides.

    Li, Ning / El Zahar, N M / Saad, Jack G / van der Hage, Erik R E / Bartlett, Michael G

    Journal of chromatography. A

    2018  Band 1580, Seite(n) 110–119

    Abstract: Alkylamines are commonly used to improve both chromatographic and mass spectral performance of electrospray ionization liquid chromatography mass spectrometry based methods for the analysis of oligonucleotides. Recently several new alkylamines have been ... ...

    Abstract Alkylamines are commonly used to improve both chromatographic and mass spectral performance of electrospray ionization liquid chromatography mass spectrometry based methods for the analysis of oligonucleotides. Recently several new alkylamines have been introduced to enhance the electrospray mass spectral response for oligonucleotides; however, the chromatographic properties of these new alkylamines have not been rigorously assessed. We have investigated the retention, peak width, resolution and general chromatographic performance of fifteen different alkylamines for the separation of a model DNA, RNA and an antisense therapeutic oligonucleotide. Eleven of the fifteen alkylamines were shown to provide similar chromatographic performance across all three classes of oligonucleotides. Based on these findings, a model for the mechanism of retention of oligonucleotides using alkylamines and hexafluoroisopropanol mobile phases is proposed. Depending on the concentrations of alkylamines and pH adjustment, oligonucleotides can be retained by micellar chromatography and not the generally held ion-pairing mechanism. This conclusion is supported by light scattering, transmission electron microscopy and ion mobility experiments detecting three micron aggregates in the mobile phase at concentrations that are routinely used for LC-MS analysis of oligonucleotides. These aggregates are not detected at lower alkylamine concentrations where the retention mechanism follows an ion-pairing mechanism. The formation of these aggregates appears to be dependent on the pH of the mobile phase.
    Mesh-Begriff(e) Amines/chemistry ; Chemistry Techniques, Analytical/methods ; Chromatography, Liquid ; DNA/isolation & purification ; Indicators and Reagents/chemistry ; Oligonucleotides/analysis ; Oligonucleotides/isolation & purification ; Propanols/chemistry ; RNA/isolation & purification ; Spectrometry, Mass, Electrospray Ionization ; Tandem Mass Spectrometry
    Chemische Substanzen Amines ; Indicators and Reagents ; Oligonucleotides ; Propanols ; hexafluoroisopropanol (3D632GYQ50) ; RNA (63231-63-0) ; DNA (9007-49-2)
    Sprache Englisch
    Erscheinungsdatum 2018-10-25
    Erscheinungsland Netherlands
    Dokumenttyp Journal Article
    ZDB-ID 1171488-8
    ISSN 1873-3778 ; 0021-9673
    ISSN (online) 1873-3778
    ISSN 0021-9673
    DOI 10.1016/j.chroma.2018.10.040
    Datenquelle MEDical Literature Analysis and Retrieval System OnLINE

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