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Updated: Apr 9, 2026

Preparation of Homogeneous MALDI Samples for Quantitative Applications
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Minimizing the Coffee Ring Effect: Improving MALDI Dried-Drop Analysis by Sequential Spotting.

Joy L Maabadi1, Arbil Lopez1, Joseph H Holbrook1

  • 1Department of Chemistry and Biochemistry, The Ohio State University, Columbus, Ohio 43210, United States.

Journal of the American Society for Mass Spectrometry
|April 8, 2026
PubMed
Summary
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Sequential spotting in matrix-assisted laser desorption/ionization mass spectrometry (MALDI MS) reduces the "coffee ring effect". This method enhances signal intensity and minimizes analyte heterogeneity in dried-drop analyses.

Area of Science:

  • Analytical Chemistry
  • Mass Spectrometry

Background:

  • Dried-drop analysis in MALDI MS is a common technique for assessing analyte ionizability and fragmentation.
  • The "coffee ring effect" causes analyte to concentrate at the edge of dried spots, leading to inconsistent signals and "dead zones".

Purpose of the Study:

  • To investigate a novel sequential spotting technique to mitigate the "coffee ring effect" in MALDI MS.
  • To improve analyte distribution and signal consistency in dried-drop analyses.

Main Methods:

  • Sequential spotting of sample solutions at reduced volumes (halved and quartered pipet solution levels).
  • Analysis of dried drops prepared using the sequential spotting method via MALDI MS.

Main Results:

  • The sequential spotting technique significantly reduced the diameter of dried spots.

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  • This method led to enhanced signal intensity and reduced prevalence of "sweet spots" and "dead zones".

    • Conclusions:

    • Sequential spotting is a viable method to improve analyte distribution and signal robustness in MALDI MS dried-drop analyses.
    • While increasing preparation time, this technique offers a more reliable approach for sample analysis.