Laser desorption ionization , 192 mass spectrum

Figure 5-5. A. MALDI mass spectrum of the anthocyanin pigments in the grape variety Marechal Foch. This figure is from the article Matrix-assisted laser desorption ionization mass spectrometry analysis of grape anthocyanins , Am. J. Enol. Vitic. 50 199-203 by Sugui, J. A., Wood, K. V., Yang, Z., Bonham, C. C. and Nicholson, R. L. 1999. Reprinted by permission of the American Society for Enology and Viticulture. B. MALDI mass spectrum of grape anthocyanins acquired on a MALDI mass spectrometer with delayed extraction capabilities. Peak identities are discussed in the text.

Figure 9 (A) Reflector MALDI mass spectrum of an in situ digest of apo-transferrin taken from the 2D map of rat sera displayed in Figure 4, which were alkylated with do-acrylamide and ds-acrylamide and mixed in a 30/70% ratio. (B) and (C) are two short intervals taken from (A), and are associated with the two indicated peptide sequences. (Reproduced from Gehanne S, Cecconi D, Carboni L, et al. (2002) Quantitative analysis of two-dimensional gel-separated proteins using isotopically marked alkylating agents and matrix-assisted laser desorption/ionization mass spectrometry. Rapid Communications in Mass Spectrometry 16 1692-1698.)...

Agapito et al. ° applied fuzzy logic to an array of semiconductor gas sensors to analyze different atmospheres for the different gases and Otto et al. developed a scheme based on the principles of fuzzy logic that makes use of various pieces of information available either from spectroscopic knowledge or from the particular spectrum that requires evaluation. A fuzzy expert system has also been successfully developed for the automated qualitative and semiquantitative interpretation of X-ray diffraction spectra/ automation of matrix-assisted laser desorption-ionization mass spectrometry (MALDI), and for polymer analysis. 

We have used accurate mass measurements obtained by matrix-assisted laser desorption ionization time-of-flight mass spectrometry (MALDI-TOFMS) to differentiate and profile saponins from M. truncatula roots. An example is provided (Fig.3.11) showing the MALDI-TOFMS spectra of a solid-phase extract of M truncatula root tissue. In this spectrum, we can identify multiple saponins. 

Example Peptides often contain sulfur from cysteine. Provided there are at least two cysteines in the peptide molecule, the sulfur can be incorporated as thiol group (SH, reduced) or sulfur bridge (S-S, oxidized). Often, both forms are contained in the same sample. At ultrahigh-resolution, the contributions of these compositions to the same nominal m/z can be distinguished. The ultrahigh-resolution matrix-assisted laser desorption/ionization (MALDI) FT-ICR mass spectrum of native and reduced [D-Pen jenkephalin gives an example of such a separation (Fig. 3.25). [39] The left expanded view shows fully resolved peaks due to and C2 isotopomers of the native and the all- C peak of the reduced compound at m/z 648. The right expansion reveals the peak of the native plus the... 

There are at least three possibile ways in which the inhibitor can bind to the active site (1) formation of a sulfide bond to a cysteine residue, with elimination of hydrogen bromide [Eq. (10)], (2) formation of a thiol ester bond with a cysteine residue at the active site [Eq. (11)], and (3) formation of a salt between the carboxyl group of the inhibitor and some basic side chain of the enzyme [Eq. (12)]. To distinguish between these three possibilities, the mass numbers of the enzyme and enzyme-inhibitor complex were measured with matrix-assisted laser desorption ionization time-of-flight mass spectrometry (MALDI). The mass number of the native AMDase was observed as 24766, which is in good agreement with the calculated value, 24734. An aqueous solution of a-bromo-phenylacetic acid was added to the enzyme solution, and the mass spectrum of the complex was measured after 10 minutes. The peak is observed at mass number 24967. If the inhibitor and the enzyme bind to form a sulfide with elimination of HBr, the mass number should be 24868, which is smaller by about one... 

Figure 3-11 Matrix-assisted laser desorption / ionization time-of-flight (MALDI-TOF) mass spectrum of bovine erythrocyte Cu-Zn superoxide dismutase averaged over ten shots with background smoothing. One-half pi of solution containing 10 pmol of the enzyme in 5 mM ammonium bicarbonate was mixed with 0.5 pi of 50 mM a-cyanohydroxycinnamic acid dissolved in 30% (v / v) of acetoni-trile-0.1% (v / v) of trifluoroacetic acid. The mixture was dried at 37° C before analysis. The spectrum shows a dimer of molecular mass of 31,388 Da, singly charged and doubly charged molecular ions at 15,716, and 7870 Da, respectively. The unidentified ion at mass 8095.6 may represent an adduct of the matrix with the doubly charged molecular ion. Courtesy of Louisa Tabatabai.

Fig. 1. Matrix-assisted laser desorption/ionization (MALDI)-time-of-flight (TOF) spectrum of a trypsin-digested one-dimensional gel band. Peaks are labeled with their monoisotopic masses. Note that these are not the masses of the peptides, but of the peptide (pseudo)molecular ions. In MALDI spectra, peptide molecular ions arise predominantly through the addition of a proton to the peptide, giving a mass increase of 1.007 Da. The molecular ions are usually denoted as MH+ or [M+H]+.

Since the signals are very short, simultaneous detection analysers or time-of-flight analysers are required. The probability of obtaining a useful mass spectrum depends critically on the specific physical proprieties of the analyte (e.g. photoabsorption, volatility, etc.). Furthermore, the produced ions are almost always fragmentation products of the original molecule if its mass is above approximately 500 Da. This situation changed dramatically with the development of matrix-assisted laser desorption ionization (MALDI) [17,18]. 

Since its discovery in 1987, matrix assisted laser desorption/ionization time of flight mass spectrometry (MALDI-TOF MS) has become a common technique in the mass spectral analysis of biopolymers (1, 2). Its ease of operation, theoretically unlimited mass range, and ability to acquire an entire mass spectrum without scanning make the technique an excellent method to analyze high mass biopolymers. Combining such advantages with the capability of analyzing sub-picomole quantities of biopolymers makes MALDI-TOF MS extremely useful for routine mass analysis. 

---