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Positive ion formation in the ultraviolet matrix-assisted laser desorption / ionization analysis of oligonucleotides by using 2,5-dihydroxybenzoic acid

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Journal of the American Society for Mass Spectrometry

Abstract

The development of matrix-assisted laser desorption/ionization (MALDI) mass spectrometry and its demonstrated performance with large proteins has generated substantial interest in utilizing this technique as an alternative to gel electrophoresis for DNA sequence analysis. However, a lack of understanding of the desorption and ionization processes has greatly hampered advances in this field. This article explores the formation of positively charged oligonucleotides in UV (355-nm) MALDI analysis by using the matrix 2,5-dihydroxybenzoic acid. Whereas substantial fragmentation is observed in the positive-ion mode by using the short oligomer d(TAGGT), no fragmentation is evident in the negative-ion mode under identical conditions. The fragmentation products are consistent with a previously published model in which base protonation initiates base loss, which leads to subsequent cleavage of the phosphodiester backbone. Several polydeoxythymidilic acids containing modified nucleosides were used to investigate positive-ion formation. The results support the hypothesis that positive ions are formed by protonation of the nucleobases. Because base protonation initiates base loss, fragmentation is intrinsic to positive-ion formation in the MALDI analysis of oligonucleotides. This result explains the dramatic difference in fragmentation observed in positive-ion compared to negative-ion UV-MALDI mass spectra of oligonucleotides.

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Authors and Affiliations

  1. Department of Chemistry, University of Wisconsin, Madison, Wisconsin, USA

    Wei Tang, Christine M. Nelson, Lin Zhu & Lloyd M. Smith

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  1. Wei Tang
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  2. Christine M. Nelson
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  3. Lin Zhu
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  4. Lloyd M. Smith
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Tang, W., Nelson, C.M., Zhu, L. et al. Positive ion formation in the ultraviolet matrix-assisted laser desorption / ionization analysis of oligonucleotides by using 2,5-dihydroxybenzoic acid. J Am Soc Mass Spectrom 8, 218–224 (1997). https://doi.org/10.1016/S1044-0305(96)00237-1

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  • DOI: https://doi.org/10.1016/S1044-0305(96)00237-1

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