Continuous flow reduced-pressure infrared laser desorption/ionization mass spectrometry

Rationale: Continuous flow ionization methods using infrared (IR) lasers have several favorable characteristics, including ionization without any additional matrices and tolerance to contaminants such as detergents and buffer salts. However, poor sensitivity due to low ion-transfer efficiency from the sample plate to the inlet capillary of the mass spectrometer under atmospheric pressure remains a serious problem. Methods: We developed a new continuous flow IR laser desorption/ionization (IR-LDI) method using a frit plate and wavelength-tunable mid-IR laser with an optical parametric oscillator. Continuous flow samples were directly injected into the ion source without any additional matrices. The ion source was covered with a decompression chamber, and could vary the pressure of the ion source from 21 to 101 kPa. Results: Reduction of the pressure of the IR-LDI source from 101 to 71 kPa increased the signal intensity for the [M + H](+) ion of angiotensin II by 1.8-fold. On the other hand, the ion signal intensity was reduced at pressures lower than 71 kPa. It became clear that reducing pressure was more effective when ionization occurred with lower laser pulse energy and lower ion source temperature. In addition, signal intensities for the [M + 2H](2+) and [M + 3H](3+) ions of insulin were also increased, by 1.4-fold and 1.1-fold, respectively, upon reduction of the pressure to 91 and 81 kPa. Conclusions: Although many studies have described IR-LDI using a differential pumping mass spectrometer, the optimal pressure of the ion source has never been investigated. We found that a slight reduction in pressure enhances sensitivity. This knowledge may be applicable to a number of ambient ionization methods using IR lasers.