Christoph R. Gebhardt

Impact dynamics of molecular clusters on surfaces: Fragmentation patterns and anisotropic effects

The fragmentation dynamics of (H2O)1032 clusters colliding with a repulsive surface at incident velocities of 1753 m/s and 2909 m/s, corresponding to kinetic energies of 0.5 and 1.5 times the cluster binding energy, has been examined in a classical molecular dynamics simulations study. The results show a large anisotropy in the energy redistribution inside the cluster upon impact, which leads to asymmetric fragmentation, starting in the leading part of the cluster. The low-mass region of the fragment size distribution can be described by a power law with an exponent close to −1.6, and the range of this region increases with increasing incident velocity. The formed fragments have rather uniform internal temperatures close to the standard boiling point of water, but the translational energy of the monomers formed upon collision is much larger, pointing at the asymmetric energy distribution inside the cluster. The angular distributions of fragment mass and fragment kinetic energy peak at grazing exit angles....

Matrix-Free Formation of Gas-Phase Biomolecular Ions by Soft Cluster-Induced Desorption†

All in a ball: Neutral molecular clusters consisting of a few thousand molecules can be seen as tiny snow balls; if they are thrown fast enough onto a surface, they are able to pick up biomolecules such as insulin from that surface. Since they break down and evaporate during and after the collision, bare biomolecular ions are available for mass spectrometry after such an energetic throw.

Desorption/ionization induced by neutral cluster impact as a soft and efficient ionization source for ion trap mass spectrometry of biomolecules.

RATIONALE Desorption and ionization induced by neutral clusters (DINeC) using SO2 as cluster constituents was previously shown to produce clear and fragmentation-free spectra with low background from samples prepared with standard oligopeptides. Here we demonstrate a more general applicability of this method based on examples from different classes of (bio-)molecules. In order to make better use of the ions generated during the millisecond cluster-pulse, the DINeC source was combined with an ion trap mass spectrometer. METHODS Desorption and ionization was induced by neutral SO2 clusters with a mean size of 10(3) to 10(4) molecules seeded in a pulsed He beam. The desorbed ions were accumulated in an ion trap over the whole pulse duration prior to mass spectrometric analysis. Samples were prepared by simply drop casting the respective aqueous solution of biomolecules on Si/SiO2 substrates. RESULTS Clear and fragmentation-free spectra of oligopeptides were detected in single pulse operation mode. The very soft nature of the desorption process was demonstrated for phosphopeptides. DINeC spectra from bovine serum albumin samples after tryptic digest led to a clear identification of the original sequence using mass fingerprinting analysis. MS(n) capability was illustrated with two types of rhodamine dyes. CONCLUSIONS Desorption and ionization induced by neutral clusters can efficiently be combined with ion trap mass spectrometry since the pulse width and repetition rate of a typical pulsed cluster beam correspond well to the discontinuous accumulation time as well as the spectral rate of the ion trap. Clear mass spectra were obtained with such a setup for a variety of biosamples demonstrating the wider applicability of the DINeC process.

A comparative study of cluster-surface collisions: Molecular-dynamics simulations of (H2O)1000 and (SO2)1000

A classical molecular-dynamics study of (H2O)1000 and (SO2)1000 clusters impacting with velocities between 6 x 10(2) and 8 x 10(3) ms at normal incidence on a repulsive target is presented. Using the ratio of total kinetic energy to total binding energy of the cluster as a scaling parameter, a general description of the fragmentation dynamics as well as the final fragment size distributions is achieved for the different systems. With increasing ratio, the angular distribution of the emitted monomers rapidly shifts from isotropic to anisotropic. At the highest investigated velocities, a tendency to recover more isotropic distributions is observed. Comparable transient compression of the impacting cluster is reached, on the other hand, for the same, unscaled collision velocities in both systems. For both H2O and SO2 the obtained internal temperatures of the cluster fragments are found to be independent of impact energy and close to the boiling temperature of the respective systems.

Quantification of the ionization probability during desorption/ionization of oligopeptides induced by neutral cluster impact

RATIONALE Desorption-and-ionization induced by neutral cluster impact is a soft and matrix-free method, which leads to the formation of free ions of oligopeptides and smaller proteins without fragmentation. As a prerequisite for its successful application in bioanalytics, especially with respect to sensitivity, the ionization efficiency, i.e., the ion-to-neutral ratio of the desorbing molecules, was determined. METHODS Neutral SO2 clusters of 10(3) to 10(4) molecules in size were seeded in a pulsed He beam and used to desorb and ionize oligopeptides by means of cluster surface impact. The samples were prepared by drop casting a well-defined amount of substance on the substrate surface; the desorbing ions were identified by means of time-of-flight mass spectrometry. Furthermore, the ion current leaving the surface was determined for positive ions, which predominate in the investigated oligopeptides. RESULTS For angiotensin II, bradykinin (1-7), and adrenocorticotropic hormone (34-39), the number of ions desorbed from the respective samples was compared with the amount of substance applied on the substrate. Assuming that all biomolecules were desorbed during the experiment, the ion-to-neutral ratio or ionization efficiency η was determined. For the tested molecules, values of η between 0.5% and 3% were observed; the substrate material and the total amount of substance applied were shown to have a minor effect on the results. CONCLUSIONS The ion-to-neutral ratio in desorption/ionization of oligopeptides induced by neutral cluster impact was determined to be of the order of 10(-3) to 10(-2). The soft and matrix-free nature of the method in combination with this value of η might be interesting for applications in bioanalytics.

Mass spectrometry of oligopeptides in the presence of large amounts of alkali halides using desorption/ionization induced by neutral cluster impact

Oligopeptides in the presence of large amounts of salt were desorbed and ionized using desorption/ionization induced by neutral clusters (DINeC) for further analysis by means of mass spectrometry (MS). Using oligopeptides in alkali halide solutions as a model system, DINeC was shown to yield clear and fragmentation free mass spectra of the biomolecules even from environments with a large excess of salt. The results were traced back to a phase separation between salt and biomolecules during sample preparation. The ratio between alkali metal complexes [M+A](+) and bare biomolecules [M+H](+) was controlled using different preparation schemes. DINeC was applied to the products of a tryptic digest of bovine serum albumin in the presence of sodium chloride; the results of a mass fingerprint analysis did not show a major difference for the spectra with and without salt in the original solution. The metal-ion/peptide interaction was further investigated by means of tandem-MS.

Influence of the cluster constituents’ reactivity on the desorption/ionization process induced by neutral SO2 clusters

The influence of the chemical nature of the cluster constituents on the desorption/ionization process was investigated for desorption/ionization induced by neutral SO2 clusters (DINeC). The polar clusters act as a transient matrix in which the desorbed analyte molecules are dissolved during the desorption process. For drop-cast samples, the desorption/ionization efficiency was found to be largely independent of the pH value of the initial solution the samples were prepared from; positive ions were almost always dominant and no multiply charged negative ions were observed. The results were traced back to the interaction of SO2 with water present in the samples. Both H/D exchange experiments and surface charge measurements showed that SO2 from the cluster beam interacts with water on and in the sample forming sulfurous acid. The latter then acts as an efficient proton supply leading to an enhanced ionization efficiency. The results demonstrate the possibility to control the ionization efficiency when using reactive cluster constituents in desorption-based ionization methods such as DINeC and cluster-based secondary ion mass spectrometry.

Real-Time Investigation of the H/D Exchange Kinetics of Porphyrins and Oligopeptides by Means of Neutral Cluster Induced Desorption/ionization Mass Spectrometry

The kinetics of the H/D exchange reaction in angiotensin II, hexaglycine (Gly6), Co(II)tetra(3-carboxyphenyl)porphyrin, and tetra(4-carboxyphenyl)porphyrin were followed in real time by mass spectrometry employing desorption/ionization induced by neutral SO2 clusters. The change of the isotope patterns with increasing degree of deuteration was recorded as a function of D2O exposure and the underlying H/D exchange kinetics, i.e., the dependence of the different degrees of deuteration on time, were deduced. The results were modeled by means of Monte Carlo simulations taking into account different reaction constants for the H/D exchange reaction at different functional groups. In the case of the investigated porphyrins, the rate constants were directly assigned to the functional groups involved; in the case of the peptides, reaction at the explicit functional groups and the backbone chain of the molecules could be discriminated.