A new approach to data reduction and evaluation in high-resolution time-of-flight mass spectrometry using a time-to-digital convertor data-recording system.

A new effective and robust approach to the detection of incompletely resolved peaks, and evaluation of their parameters in high-resolution time-of-flight mass spectra for time-to-digital convertor (TDC) data acquisition mode, is described. The method is based on fast construction of a smoothed continuous curve that approximates the initial data (transformed to a constant relative width of time intervals for ion counting) with respect to precision of measurements. The first derivative of this curve is used for correction of skewness of the peak shape as far as possible. A contribution of the second derivative is subtracted from the smoothed curve for better resolution of partially resolved peaks. The comparison of local maxima of this resulting final curve with those for the initial smoothed curve allows reliable detection of the peaks and to test whether or not they are spoiled by overlapping. Ion counting performed by TDC gives an opportunity to estimate standard deviations of peak locations and their intensities. These values proved to be close to theoretically minimal standard deviations for these parameters for single fully resolved peaks. Thus, estimates of the main parameters of mass peaks by the described method are close to statistically efficient estimators for these parameters.

The solvent shells of cluster ions produced by direct electric field extraction from glycerol/water mixtures.

Electric field extraction of gaseous negative ions directly from water/glycerol solutions by use of a track membrane technique was investigated. The distributions of numbers of solvent molecules in the extracted cluster ions for different compounds were obtained. It is shown that the extraction mechanism is a direct field-stimulated evaporation of cluster ions from liquid, with a subsequent loss of several solvent molecules in the vacuum. For relatively simple ions a good correspondence of results was obtained with a continuous medium model. It was found that the number of solvent molecules in a cluster shell, for more complicated ions such as amino acids, is significantly greater than that for halide ions or ions of simple organic acids. An increase in the number of solvent molecules in the case of amino acid negative ions is rationalized in terms of the existence of several charged groups, each of which gives an additional contribution to the cluster shell.

Electrospray mass spectrometric study of melittin trypsinolysis by a kinetic approach.

The kinetics of tryptic digestion of melittin was studied by combined electrospray ionization time-of-flight mass spectrometry and high-performance liquid chromatography. The ratios of the kinetic constants for cleavage of the peptide bonds that are susceptible to trypsin action were determined. It is shown that trypsin does not manifest affinity for the hydrolysis of the peptide bonds inside the Arg,Lys cluster series as efficiently as it cleaves the peptide at the separately localized Lys residue. This feature demonstrates clearly the advantage of the kinetic approach to tryptic mapping of proteins. The kinetic approach allows the determination of not only discrete structural segments in protein structure but also their relative locations and their amino acid sequences. Using the melittin digests and some artificially prepared amino acids and dipeptides mixtures as models, it is shown that the presence and nature of basic amino acids predetermines the charge states of the molecules analyzed by electrospray but not the yields of their ions. The aliphatic parts of the molecules seem to be more important in determining the actual ion yields.