Improvement of resolution, mass accuracy, and reproducibility in reflected mode DE-MALDI-TOF analysis of DNA using fast evaporation - Overlayer sample preparations

John M. Koomen, William K. Russell, Justin M. Hettick, David H. Russell

Research output: Contribution to journalArticle

35 Scopus citations

Abstract

DNA analysis by matrix-assisted laser desorption/ionization (MALDI) mass spectrometry is hindered by two processes: alkali metal adduction and fragmentation of the intact ionized molecule. The adverse effects of both processes can be reduced by adding ammonium ion salts or compounds such as fructose to the sample preparations. Matrix additives improve sensitivity and resolution of DNA analysis by MALDI. In addition, spot-to-spot reproducibility, resolution, and mass accuracy for DNA oligonucleotides (≤ 12 mer) can be improved by the use of overlayer sample preparations with matrixes that have low aqueous solubilities, such as α-cyano-4-hydroxycinnamic acid, ferulic acid, and 2,4,6-trihydroxyacetophenone. For example, resolution for 5-12-mer oligonucleofides is greater than 7000 using overlayer matrix preparations and mass accuracy values are well below 20 ppm. In addition to these methods, a new method for analyzing DNA in positive ion mode is reported using acidified 3-hydroxypicolinic acid. This method does not lose sensitivity for higher mass oligonucleotides as quickly as overlayer methods, and spectra retain > 6000 resolution and mass accuracies of ~20 ppm between different overlayer depositions.

Original languageEnglish (US)
Pages (from-to)3860-3866
Number of pages7
JournalAnalytical Chemistry
Volume72
Issue number16
DOIs
StatePublished - Aug 15 2000
Externally publishedYes

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ASJC Scopus subject areas

  • Analytical Chemistry

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