Ionization mechanism of oligonucleotides in matrix-assisted laser desorption/ionization time-of-flight mass spectrometry

The ionization of nucleosides in matrix‐assisted laser desorption/ionization time‐of‐flight mass spectrometry was systematically investigated using adenine (A), thymine (T), guanine (G) and cytosine (C) with several common matrices. Experimental results of the protonation and deprotonation of the bases of A, T, G and C in the matrices 2,5‐dihydroxybenzoic acid (2,5‐DHB), α‐cyano‐4‐hydroxycinnamic acid (α‐CHCA) and 3‐hydroxypicolinic acid (3‐HPA) provide an insight into the ionization mechanism of oligonucleotides in MALDI. It was found that the low ion signal from DNA in poly‐G in MALDI as reported in earlier work could be attributed to the fact that the base of G is difficult to ionize. Our results suggest that the ionization of DNA in MALDI is dominated by the protonation and deprotonation of bases and it is basically independent of the backbone of DNA. Both the protonation and deprotonation are strongly structure dependent. The protonation is dominated by pre‐protonation before laser ablation, while the deprotonation is controlled by the thermal reaction. Copyright © 2000 John Wiley & Sons, Ltd.