A tunable Cr:LiSAF laser-pumped optical parametric oscillator was used for mid-infrared matrix-assisted laser desorption/ionization (MALDI) mass spectrometry experiments. The mass spectra of substance P, bovine insulin and pd(T)10 in the 2.88-2.96 microns range showed excellent single shot signal quality (signal-to-noise-ratio, resolution) but poor shot-to-shot reproducibility. The reproducibility is expected to improve with more stable laser design. No correlation was found between the absorption spectrum of the matrix and the MALDI response.