This investigation concerns the initial chemical reactions that affect the ionization of matrixes in matrix-assisted laser desorption/ionization (MALDI). The study focuses on the relaxations of photon energy that occur on a comparable time scale to that of ionization, in which the available laser energy is shared and the ionization condition is changed. The relaxations include fluorescence, fragmentation, and nonradiative relaxation from the excited state to the ground state. With high absorption cross section and long excited-state lifetime, photoionization of matrix plays an important role if sufficient laser energy is used. Under other conditions, thermal ionization of the molecule in the ground state is predicted to be one of the important reactions. Evidence of change in the branching ratio of initial reactions with the matrix and the excitation wavelength was obtained with α-cyano-4-hydroxycinnamic acid, sinapinic acid, 2,5-dihydroxybenzoic acid, and 2,4,6-trihydroxyacetophenone. These matrixes are studied by obtaining their mixed crystal absorption spectra, fluorescence properties, laser-induced infrared emission, and product ions. The exact ionization pathway depends on the chemical properties of matrixes and the excitation conditions. This concept may explain the diversity of experimental results observed in MALDI experiments, which provides an insight into the ensemble of chemical reactions that govern the generation of ions.

Positive and negative ions produced from matrix-assisted laser desorption/ionization (MALDI) were simultaneously measured using a newly developed dual-polarity time-of-flight mass spectrometer. This instrument is effective not only for express and comprehensive mass analysis but also for studying the ionization mechanisms of biomolecules. It comprises two identical time-of-flight mass analyzers located symmetrically about a MALDI ion source. The ion optics are arranged to be able to extract positive and negative ions synchronously with equal efficiency to each corresponding mass analyzer. Mass spectra of various proteins with molecular weights as large as that of myoglobin monomer and dimer were obtained. The spectral patterns obtained in this work are approximately mirror images with opposite polarities.

This work presents the unique features of a novel configuration of a synchronized dual-polarity time-of-flight mass spectrometer for comprehensive surface imaging. Mass spectrometry imaging of surface samples covering positive and negative ion modes is difficult due to rapid signal depletion. This limitation is overcome here by dual-polarity time-of-flight mass spectrometry (DP-TOFMS) via two separate TOF mass analyzers that are installed above a sample surface. The new instrument eliminates the polarity bias characteristic of most mass spectrometers, which is important for the analysis of samples with diverse physical and chemical properties. The experimental results show for the first time that the spatial distribution of positive and negative ions of various photolabile samples can be distinguished, including pigments and conventional matrix-assisted laser desorption/ionization samples. The different positive and negative ion distributions suggest that accurate quantitative information can only be obtained when the entire sample region is examined by DP-TOFMS, which was unfeasible in the past. Such a comprehensive diagnostic method is essential for the molecular imaging of trace compositions in delicate biological tissues, as demonstrated here with a Phyllanthus urinaria leaf that only produced ion signals in the first examination and not in the subsequent measurements.

This work demonstrates the feasibility of a novel scintillation detector with greater detection efficiency than that of chevron-type microchannel plate (MCP) detectors. The detection mechanism involves sequential conversion reactions induced by ion-surface impacts. Identical detection conditions can be utilized to monitor both positive and negative ions in mass spectrometers. The proposed detector comprises an ion beam guiding device, a negatively biased washer-shaped conversion dynode, and an aluminum-coated scintillation detector. The beam guide changes the electric field around the washer-shaped conversion dynode, and it allows the primary and secondary ions to propagate toward the scintillation phosphor and the conversion dynode, respectively. The detection is achieved by the detection of electron-induced luminescence on a phosphor. The amplification efficiency of this bipolar ion detector increases as the conversion dynode voltage increases. For ions with a mass-to-charge ratio of up to 90 000, the sensitivity of the BID is 1.4-14.4 times that of the MCP. Further improvement of the sensitivity can be achieved by increasing the conversion dynode voltage or the ion acceleration voltage. Results of this study demonstrate that this detector is a promising alternative for efficient ion detection.