Near-ultraviolet-induced matrix-assisted laser desorption/ionization as a function of wavelength

The matrix-assisted laser desorption/ionization (MALDI) effect for the ablation of polypeptide ions was studied as a function of wavelength in the range 360–450 nm, using a tunable titanium:sapphire laser and the matrices (trans)α-cyano-4-hydroxycinnamic acid, (trans)3,5-dimethoxy-4-hydroxycinnamic acid, and (trans)β-indoleacrylic acid. The results demonstrated that the qualitative aspects of the MALDI effect remained largely unchanged as a function of wavelength until the matrix crystal’s absorption coefficient approached 6 × 10⁴ cm⁻¹. Wavelengths at which the absorption coefficient was lower than this value did not produce observable polypeptide ions. In the range of wavelengths where polypeptide ions could be observed, the effect was generated only when the average energy absorbed in the 10-nm layer immediately below the crystal surface was >8 kJ/cm³ regardless of the total amount of energy absorbed in the remainder of the crystal. It was demonstrated that on the basis of the other results in this paper and those in the literature that the mechanism that generates polypeptide ions cannot be either photochemically induced or the same mechanism that gives rise to matrix ions. A model for ionization is proposed that decouples matrix and protein ionization in a simple manner that has direct analogy to the behavior of ionic solutes in polar solvents. The model satisfies the currently available experimental evidence and unifies the results obtained by MALDI at all wavelengths.