MALDI mass spectrometry Matrix-assisted laser desorption

Matrix-assisted laser desorption mass spectrometry (MALDI-MS) is, after electrospray ionization (ESI), the second most commonly used method for ionization of biomolecules in mass spectrometry. Samples are mixed with a UV-absorbing matrix substance and are air-dried on a metal target. Ionization and desorption of intact molecular ions are performed using a UV laser pulse. 

Concentration detection limits in CE-MS with the ESI interface are similar to those with UV detection. Sample sensitivity can be improved by using ion-trapping or time-of-flight (TOE) mass spectrometers. MS analysis can also be performed off-line, after appropriate sample collection, using plasma desorption-mass spectrometry (PD-MS) or matrix-assisted laser desorption-mass spectrometry (MALDI-MS). 

MALDI-MS Matrix-assisted laser desorption mass spectrometry... 

Recent advances in mass spectrometry (MS) technology have provided researchers with an unparalleled ability to identify the types and patterns of secondary biochemical modifications found on proteins in living cells. Matrix-assisted laser desorption/ionization-MS (MALDI-MS) analyses have shown, for example, that HMGA proteins in vivo are simultaneously subject to complex patterns of phosphorylation, acetylation and methylation and that, within the same cell type, different isoforms of these proteins can exhibit quite different modification patterns [33]. Furthermore, these in vivo modifications have been demonstrated to markedly alter the binding affinity of HMGA proteins for both DNA and chromatin substrates in vitro [33]. Nevertheless, due to their number and complexity, it has been difficult to determine the actual biological function(s) played by these biochemical modifications in living cells. 

Mass spectrometry is a powerful qualitative and quantitative analytical tool that is used to assess the molecular mass and primary amino acid sequence of peptides and proteins. Technical advancements in mass spectrometry have resulted in the development of matrix-assisted laser desorption/ion-ization (MALDI) and electrospray ionization techniques that allow sequencing and mass determination of picomole quantities of proteins with masses greater than 100kDa (see Chapter 7). A time-of flight mass spectrometer is used to detect the small quantities of ions that are produced by MALDI. In this type of spectrometer, ions are accelerated in an electrical field and allowed to drift to a detector. The mass of the ion is calculated from the time it takes to reach the detector. To measure the masses of proteins in a mixture or to produce a peptide map of a proteolytic digest, from 0.5 to 2.0 p.L of sample is dried on the tip of tlie sample probe, which is then introduced into tire spectrometer for analysis. With this technique, proteins located on the surfaces of cells are selectively ionized and analyzed. 

Mass spectrometers see Chapter 7) coupled with gas and liquid chromatographs (see Chapter 6) afe powerful qualitative and quantitative analytical tools that are widely used to measure hormones. Technical advancements in mass spectrometry have resulted, in the, development of matrix-assisted laser desorption/ioiiizati,6n (MALDI) and electro-spray ionization techiiiques that allow siequencing of peptides and mass determination of piconiole quantities of analytes. 

New ionization methods and mass analyzers have extended the applicability and overall sensitivity of to mass spectrometry to macromolecular targets. Mass spectrometry can be used to directly study the covalent and noncovalent interactions of drug molecules and biomolecule targets. Inhibitors that bind irreversibly or covalently to a target are easily studied by use of MS because the covalent complex survives intact in the gas phase. MS is combined with other proteomic technologies as well for use in drug discovery e.g., matrix-assisted laser desorption ionization MS (MALDI-MS) and electrospray ionization (ESI). 

Soft ionization MS techniques [9] like electrospray ionization (ESI) and soft laser desorption, often known as matrix-assisted laser desorption/ioniza-tion (MALDI), facilitated the polymer analyses over the last years. The advantage of the soft ionization techniques is the transformation of dissolved liquid or solid sample into the gas phase, where no change in the molecular composition/structure of the sample will be induced, while hard ionization in mass spectrometry (e.g., electron ionization (El) or fast atom bombardment (FAB)) preferentially destroys the chemical and molecular structure into fragments prior to the detection of the molar mass fragments of the sample by mass spectrometry. 

Mass spectrometry currently has assumed a central role in protein sequencing. This development has been possible with the introduction of two highly sensitive ionization techniques electrospray ionization (ESI) and matrix-assisted laser desorption and ionization (MALDI) and the advent of improved instrumentation capable of high-mass and high-sensitivity detection. Currently, biopolymers with a molecular mass over 100,000 Da are analyzed routinely. In the past, fast atom bombardment (FAB) [6,7] and Cf plasma desorption (PD) ionization [8] also played a limited role in protein sequencing. Mass spectrometry now has assumed... 

Chapter 6, titled Selection of Ionization Methods of Analytes in the TLC-MS Techniques provides an overview of mass spectrometric techniques that can be coupled with TLC and act as specific detectors in this hyphenated approach. The mass spectrometric techniques discussed in this chapter are secondary mass spectrometry (SIMS), liquid secondary ion mass spectrometry (LSIMS), fast atom bombardment (FAB), matrix-assisted laser desorption/ionization (MALDI), atmospheric pressure matrix-assisted laser desorption/ionization (AP-MALDI), electrospray ionization (ESI), desorption electrospray ionization (DESI), electrospry-assisted laser desorption/ionization (ELDI), easy ambient sonic spray ionization (EASI), direct analysis in real time (DART), laser-induced acoustic desorption/electrospray ionization (LIAD/ESI), plasma-assisted multiwavelength laser desorption/ionization (PAMLDI), atmospheric-pressure chemical ionization (APCI), and dielectric barrier discharge ionization (DBDI). For the sake of illustration, the authors introduce practical examples of implementing TLC separations with detection carried out by means of individual mass spectrometric techniques for the systematically arranged compounds belonging to different chemical classes. 

The major reason for this increase has been the use of matrix-assisted laser desorption/ionisation-MS (MALDI-MS) for numerous polymer applications. MALDI is by no means the only mass spectral method that is useful for polymer analysis, but it has provided the impetus to get polymer people interested in what mass spectrometry can do. 

In addition to these characterization tools, surface analytical techniques such as surface matrix-assisted laser desorption ionization (Surface MALDI) mass spectrometry, Rutherford backscattering spectroscopy (RBS), and near-edge X-ray absorption fine stmcture spectroscopy (NEXAFS) are used to obtain structural and chemical details about surface thin films. Surface MALDI, also known as MALDI-ToF MS (see Section 5.4.2), offers high mass resolution for analyzing surface films and molecular layers using the m/z of various ions generated from the sample surface (mixed with an... 

Until about the 1990s, visible light played little intrinsic part in the development of mainstream mass spectrometry for analysis, but, more recently, lasers have become very important as ionization and ablation sources, particularly for polar organic substances (matrix-assisted laser desorption ionization, MALDI) and intractable solids (isotope analysis), respectively. 

Laser-desorption mass spectrometry (LDMS) or matrix-assisted laser desorption ionization (MALDI) coupled to a time-of-flight analyzer produces protonated or deprotonated molecular ion clusters for peptides and proteins up to masses of several thousand. 

MALDI = matrix assisted laser desorption, ftms = Fourier transform mass spectrometry TOF = time of flight. 

With the identities and amounts of amino acids known, the peptide is sequenced to find out in what order the amino acids are linked together. Much peptide sequencing is now done by mass spectrometry, using either electrospray ionization (ESI) or matrix-assisted laser desorption ionization (MALDI) linked to a time-of-flight (TOF) mass analyzer, as described in Section 12.4. Also in common use is a chemical method of peptide sequencing called the Edman degradation. 

MALDI (Section 12.4) Matrix-assisted laser desorption ionization a mild method for ionizing a molecule so that fragmentation is minimized during mass spectrometry. 

Two relatively new techniques, matrix assisted laser desorption ionization-lime of flight mass spectrometry (MALDI-TOF) and electrospray ionization (FS1), offer new possibilities for analysis of polymers with molecular weights in the tens of thousands. PS molecular weights as high as 1.5 million have been determined by MALDI-TOF. Recent reviews on the application of these techniques to synthetic polymers include those by Ilantoif54 and Nielen.555 The methods have been much used to provide evidence for initiation and termination mechanisms in various forms of living and controlled radical polymerization.550 Some examples of the application of MALDI-TOF and ESI in end group determination are provided in Table 3.12. The table is not intended to be a comprehensive survey. 

The molecular weights and molecular weight distributions (MWD) of phenolic oligomers have been evaluated using gel permeation chromatography (GPC),23,24 NMR spectroscopy,25 vapor pressure osmometry (VPO),26 intrinsic viscosity,27 and more recently matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF MS).28... 

Maleamic acid, cyclization of, 293 Maleic anhydride, 59 Maleimido azine, 307 Manganese diacetate catalysts, 71 Mark-Houwink-Sakurada equation, 57 Material safety data sheets (MSDSs), 246 Matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOFMS), 385, 388 McGrath, J. E., 327 MDI isomers, 210 MDIs. See Methylene diphenyl diisocyanates (MDIs)... 

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