MALDI sensitivity

A MALDI-MS study of DNA analysis using 2,4,6-THAP and 2,3,4-trihydroxyaceto-phenone (2,3,4-THAP), separately and in combination, was performed. The results showed that a mixture of 2,3,4-THAP, 2,4,6-THAP, and ammonium citrate with molar ratios of 2 1 1 serves as a good matrix for the detection of DNA, especially for samples containing a small quantity of DNA such as PCR products. The resolution and shot-to-shot reproducibility using this matrix combination were better than, and the MALDI sensitivity comparable to, that obtained when using 3-HPA, PA, and ammonium citrate matrix (9 1 1). 

The hybrid can be used with El, Cl, FI, FD, LSIMS, APCI, ES, and MALDI ionization/inlet systems. The nature of the hybrid leads to high sensitivity in both MS and MS/MS modes, and there is rapid switching between the two. The combination is particularly useful for biochemical and environmental analyses because of its high sensitivity and the ease of obtaining MS/MS structural information from very small amounts of material. The structural information can be controlled by operating the gas cell at high or low collision energies. 

The mass spectrometer is a mass-flow sensitive device, which means that the signal is proportional to the mass flow dm/dl of the analyte, i.e. the concentration times the flow-rate. It is only now possible to realise the high (theoretically unlimited) mass range and the high-sensitivity multichannel recording capabilities that were anticipated many years ago. Of considerable interest to the problem of polymer/additive deformulation are some of the latest developments in mass spectrometry, namely atmospheric pressure ionisation (API), and the revival of time-of-flight spectrometers (allowing GC-ToFMS, MALDI-ToFMS, etc.). 

Tables 6.27 and 6.31 show the main characteristics of ToF-MS. ToF-MS shows an optimum combination of resolution and sensitivity. ToF-MS instruments provide up to 40000 spectra s-1, a mass range exceeding 100000 (in principle unlimited), a resolution of 5000, and peak widths as short as 200 ms. This is better than quadruples and most ion traps can handle. Unlike the quadrupole-type instrument, the detector is detecting every introduced ion (high duty factor). This leads to a 20- to 100-times increase in sensitivity, compared to QMS used in scan mode. The mass range increases quadratically with the time range that is recorded. Only the ion source and detector impose the limits on the mass range. Mass accuracy in ToF-MS is sufficient to gain access to the elemental composition of a molecule. A single point is sufficient for the mass calibration of the instrument. ToF mass spectra are commonly calibrated using two known species, aluminium (27 Da) and coronene (300 Da). ToF is well established in combination with quite different ion sources like in SIMS, MALDI and ESI.

Da of the monoisotopic masses, high sensitivity (down to 20fmolp,L 1), and the absence of any fragmentation, are important advantages for a L-ToF system. The main characteristics of MALDI-ToFMS, as applied directly to polymer/additive dissolutions, are summarised in Table 9.7. 

Applications MALDI-ToFMS is at its best as a rapid screening technique for quick identification of known additives. However, this screening is rendered slightly more complicated by the fact that MALDI-ToFMS spectra of pure additives and of additives in the presence of excess macromolecules are not always identical (matrix effect) [55]. For unknown additives, the relation MALDI-ToFMS spectrum-chemical structure is not easily established, and the use of FD or MALDI-MS/MS is then needed. As MALDI-MS shows a sensitivity difference for the various additives, it cannot easily quantify them unless the analytes are very similar. For differentiation of additives with the same mass number (e.g. Tinuvin 315 and Cyasorb UV3638 with m/z = 368) high resolution is required, as provided by delayed extraction MALDI-ToFMS. 

Wang et al. also addressed the mass spectral reproducibility. They conducted a carefully controlled interlaboratory experiment where the effects of a number of parameters were systematically investigated.22 They demonstrated that nearly identical spectra could be obtained in carefully controlled experiments. Minor variations in the sample/matrix preparation procedures for MALDI and in the experimental conditions used for bacterial protein extraction or analysis were shown to result in changes in the resulting spectra. They also noted that a subset of peaks was less sensitive to experimental variables. These ions appeared to be conserved in spectra obtained even under different experimental conditions so long as they were obtained using genetically identical bacteria. The existence of these conserved peaks helped explain... 

A few particularly novel approaches have been developed to increase the sensitivity (and specificity) involved in the detection of bacteria by MALDI. The use of avadin-biotin technology to recover biotinylated proteins from... 

The two remaining shortfalls with MALDI-MS analysis of whole bacterial cells are sensitivity and mixture analysis. The sensitivity for MALDI-MS analysis of whole-cell bacteria from our experiments and those reported by other laboratories is about 107 cells/ml. To realistically utilize MALDI MS as a tool that meets DoD detector sensitivity goals, this should be 103 cells/ml or lower. 

There are two different oligomer series present in all spectra. The oligomer series can be identified by calculating the masses of the end groups and assigning them to specific chemical structures (Pasch and Schrepp, 2003 Weidner et al., 2004). In the present example, the two species are the propionic amide-acid (R-am-ac) and the propionic amide-propionic amide polyamides (R-am-am-R). The use of MALDI-TOF MS as a structure-sensitive detector allows the resolution to be indirectly enhanced since several species coelute, as shown in Fig. 17.21. The polarity of the... 

Experimental considerations Sample preparation and data evaluation are similar to membrane osmometry. Since there is no lower cut-off as in membrane osmometry, the method is very sensitive to low molar mass impurities like residual solvent and monomers. As a consequence, the method is more suitable for oligomers and short polymers with molar masses up to (M)n 50kg/mol. Today, vapour pressure osmometry faces strong competition from mass spectrometry techniques such as matrix-assisted laser desorption ionisation mass spectrometry (MALDI-MS) [20,21]. Nevertheless, vapour pressure osmometry still has advantages in cases where fragmentation issues or molar mass-dependent desorption and ionization probabilities come into play. 

Lemaire R, Wisztorski M, Desmons A, et al. MALDI-MS direct tissue analysis of proteins improving signal sensitivity using organic treatments. Anal. Chem. 2006 78 7145-7153. 

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