Development of a new electron ionization/field ionization ion source for gas chromatography/time-of-flight mass spectrometry.

We have developed a combined EI/FI source for gas chromatography/orthogonal acceleration time-of-flight mass spectrometry (GC/oaTOFMS). In general, EI (electron ionization) and FI (field ionization) mass spectra are complementary: the EI mass spectrum contains information about fragment ions, while the FI mass spectrum contains information about molecular ions. Thus, the comparative study of EI and FI mass spectra is useful for GC/MS analyses. Unlike the conventional ion sources for FI and EI measurements, the newly developed source can be used for both measurements without breaking the ion source vacuum or changing the ion source. Therefore, the combined EI/FI source is more preferable than the conventional EI or FI ion source from the viewpoint of the reliability of measurements and facility of operation. Using the combined EI/FI source, the complementarity between EI and FI mass spectra is demonstrated experimentally with n-hexadecane (100 pg): characteristic fragment ions for the n-alkane such as m/z 43, 57, 71, and 85 are obtained in the EI mass spectrum, while only the parent peak of m/z 226 (M+) without any fragment ions is observed in the FI mass spectrum. Moreover, the field desorption (FD) measurement is also demonstrated with poly(ethylene glycol)s M600 (10 ng) and M1000 (15 ng). Signals of [M+H]+, [M+Na]+ and [M+K]+ are clearly detected in the FD mass spectra.

Denaturation of lysozyme and myoglobin in laser spray.

In laser spray, the tip of an electrospray capillary is irradiated with a continuous CO(2) laser beam. Here, we report results from a modified laser spray method that employs a relatively low laser irradiance level. With a laser power of approximately 2 W and a focal spot size ( approximately 0.3 mm), which covered the entire front surface of the electrospray capillary, the irradiance was approximately 3 x 10(3) W/cm(2). This resulted in a quiescent and smooth vaporization of aqueous solutions. This "evaporation-mode" laser spray method yielded the best results so far obtained in our laboratory with laser-irradiated electrospray, producing higher and more stable signals. The method was applied to the analysis of aqueous solutions of lysozyme and myoglobin. Mass spectra were obtained as a function of laser power from 0 W (electrospray) to approximately 2 W. The spray generated at the tip of the stainless steel capillary was observed with a CCD camera. With increase of laser power, the droplets in the spray became finer and the Taylor cone became progressively smaller. The strongest ion signals were recorded when the sample solution protruded only slightly from the tip of the capillary. A broadening of the lysozyme charge-state distribution, attributable to protein unfolding, was observed with a laser power of 2 W. No denaturation of myoglobin took place up to a laser power of 1.6 W. However, a sudden onset of denaturation was observed at 1.8 W as a broadening of the myoglobin charge distribution and the appearance of apo-myoglobin peaks. These findings demonstrate that laser spray is capable of dissociating the noncovalent complexes selectively without breaking covalent bonds.

Study on ion formation in electrospray droplet impact secondary ion mass spectrometry.

A new type of cluster secondary ion mass spectrometry (SIMS), named electrospray droplet impact (EDI), has been developed in our laboratory. In general, rather strong negative ions as well as positive ions can be generated by EDI compared with conventional SIMS. In this work, various aspects of ion formation in EDI are investigated. The Brønsted bases (proton acceptor) and acids (proton donor) mixed in the analyte samples enhanced the signal intensities of deprotonated molecules (negative ions) and protonated molecules (positive ions), respectively, for analytes. This suggests the occurrence of heterogeneous proton transfer reactions (i.e. M + M' --> [M+H](+) + [M'-H](-)) in the shockwave-heated selvedge of the colliding interface between the water droplet and the solid sample deposited on the metal substrate. EDI-SIMS shows a remarkable tolerance to the large excess of salts present in samples. The mechanism for desorption/ionization in EDI is much simpler than those for MALDI and SIMS because only very thin sample layers take part in the shockwave-heated selvedge and complicated higher-order reactions are largely suppressed.

Ion/molecule reactions in the orifice-skimmer region of an atmospheric pressure Penning ionization mass spectrometer.

Atmospheric pressure Penning ionization mass spectra of methanol were measured as functions of Ar or He gas pressure in the first vacuum chamber, the position of the skimmer, and the voltage applied between the orifice and the skimmer. When the orifice and the skimmer were coaxial with a distance of 4 mm, the distribution of CH3OH2+(CH3OH)n clusters was only weakly dependent on both Ar pressure (in the range of 19-220 Pa) and orifice-skimmer voltage (in the range of 1-45 V). The ion/molecule reaction CH3OH2+ + CH3OH --> CH3+(CH3OH) + H2O was observed in the free jet expansion, especially at high orifice-skimmer voltage values. When the orifice and the skimmer were off-centered and the distance between them was increased to 18 mm, the formation of large CH3OH2+(CH3OH)n clusters, as well as their dissociation, were seen. The endothermic proton transfer reaction, CH3+(CH3OH) + CH3OH --> CH3OH2+ + CH3OCH3, occurred at high orifice-skimmer voltage. The collision-induced dissociation of cluster ions by He gas in the first vacuum chamber was much more efficient than by Ar. These results demonstrated that the mass spectra are highly dependent on skimmer position and on orifice-skimmer voltage and that ions observed by mass spectrometry do not necessarily reflect the abundance of ions produced in the atmospheric pressure ion source.

Atmospheric-pressure Penning ionization mass spectrometry.

A preliminary study on the atmospheric-pressure Penning ionization (APP(e)I) of gaseous organic compounds with Ar* has been made. The metastable argon atoms (Ar*: 11.55 eV for (3)P(2) and 11.72 eV for (3)P(0)) were generated by the negative-mode corona discharge of atmospheric-pressure argon gas. By applying a high positive voltage (+500 to +1000 V) to the stainless steel capillary for the sample introduction (0.1 mm i.d., 0.3 mm o.d.), strong ion signals could be obtained. The ions formed were sampled through an orifice into the vacuum and mass-analyzed by an orthogonal time-of-flight mass spectrometer. The major ions formed by APP(e)I are found to be molecular-related ions for alkanes, aromatics, and oxygen-containing compounds. Because only the molecules with ionization energies less than the internal energy of Ar* are ionized, the present method will be a selective and highly sensitive interface for gas chromatography/mass spectrometry.