Mass accuracy requirements

How often an LC-MS should be calibrated depends on the mass accuracy required. For example, instrument calibration should be verified daily when performing accurate mass measurements of peptides and proteins. However, the quantitative analysis of small molecules requires less frequent calibration. 

Direct ES-MS characterization of a combinatorial library has been performed on the example of modified xanthene derivatives [30]. To circumvent the analysis of 104 to 10s possible compounds, small representative sublibraries rather than the whole library were synthesized and investigated [31].The results obtained were then extrapolated to the synthesis of the target library. A xanthene library was also used for ES-FT-ICR-MS investigation [32], A more recent contribution deals with the calculation of elemental compositions from FTICR mass spectra. FIowever,the mass accuracy required increases exponentially with the mass of the analyte [33], Therefore, elemental compositions are calculated from the exact measurement of fragment ions performing MS"-experiments [34],... 

One of the important advantages of ICPMS in problem solving is the ability to obtain a semiquantitative analysis of most elements in the periodic table in a few minutes. In addition, sub-ppb detection limits may be achieved using only a small amount of sample. This is possible because the response curve of the mass spectrometer over the relatively small mass range required for elemental analysis may be determined easily under a given set of matrix and instrument conditions. This curve can be used in conjunction with an internal or external standard to quantily within the sample. A recent study has found accuracies of 5—20% for this type of analysis. The shape of the response curve is affected by several factors. These include matrix (particularly organic components), voltages within the ion optics, and the temperature of the interffice. 

State-of-the-art TOF-SIMS instruments feature surface sensitivities well below one ppm of a mono layer, mass resolutions well above 10,000, mass accuracies in the ppm range, and lateral and depth resolutions below 100 nm and 1 nm, respectively. They can be applied to a wide variety of materials, all kinds of sample geometries, and to both conductors and insulators without requiring any sample preparation or pretreatment. TOF-SIMS combines high lateral and depth resolution with the extreme sensitivity and variety of information supplied by mass spectrometry (all elements, isotopes, molecules). This combination makes TOF-SIMS a unique technique for surface and thin film analysis, supplying information which is inaccessible by any other surface analytical technique, for example EDX, AES, or XPS. 

This strategy requires a more complex system for sample processing however, it engages the advantages of improved mass accuracy and better sensitivity that derive from working with lower mass ions. Peptide maps from protein mixtures have been demonstrated to provide reliable identifications of microorganisms that are reasonably pure.79-81... 

Zubarev, R. A. Hakansson, P. Sundqvist, B. Accuracy requirements for peptide characterization by monoisptopic molecular mass measurements. Anal. Chem. 1996,68,4060 1063. 

In LC-MS, specific ionisation conditions can be required for different types of species. This means that in LC-MS studies on extractable additives, it is necessary to use a range of experimental conditions to cover detection of all types of potential species. Depending on instrument type, it is also possible to isolate ions in complex matrices and obtain positive identifications by further unique fragmentation of these ions (by MS-MS or MSn). Quantitative methods based on this secondary ionisation can be employed. The mass accuracy of LC-MS detection systems continues to improve. Accurate mass measurement improves the certainty of identification. Advanced systems are typically offering 1-2 ppm (mass dependent) mass accuracy. 

The m/z values of peptide ions are mathematically derived from the sine wave profile by the performance of a fast Fourier transform operation. Thus, the detection of ions by FTICR is distinct from results from other MS approaches because the peptide ions are detected by their oscillation near the detection plate rather than by collision with a detector. Consequently, masses are resolved only by cyclotron frequency and not in space (sector instruments) or time (TOF analyzers). The magnetic field strength measured in Tesla correlates with the performance properties of FTICR. The instruments are very powerful and provide exquisitely high mass accuracy, mass resolution, and sensitivity—desirable properties in the analysis of complex protein mixtures. FTICR instruments are especially compatible with ESI29 but may also be used with MALDI as an ionization source.30 FTICR requires sophisticated expertise. Nevertheless, this technique is increasingly employed successfully in proteomics studies. 

A mass calibration for FTICR analyzers with superconducting magnets is very stable and is valid for many days for normal applications. Mass accuracy < 1 ppm can be obtained over a fairly wide mass range. Unique elemental composition can be determined for masses over 800 Da [262]. Recently, 0.1 ppm mass accuracy, which required a mass resolving power >300,000, has been achieved for several thousand peaks by a 14.5 T instrument [263] and commercial instruments with mass accuracy <0.2 ppm are available. As with the orbitrap (see Section 2.2.5) the frequency is... 

Isotopic patterns provide a prime source of such additional information. Combining the information from accurate mass data and experimental peak intensities with calculated isotopic patterns allows to significantly reduce the number of potential elemental compositions of a particular ion. [31] Otherwise, even at an extremely high mass accuracy of 1 ppm the elemental composition of peptides, for example, can only be uniquely identified up to about 800 u, i.e., an error of less than 0.8 mmu is required even if only C, H, N, O and S are allowed. [27,32,33]... 

The number of decimal places one should employ in mass calculations depends on the purpose they are used for. In the m/z range up to about 500 u, the use of isotopic mass with four decimal places will provide sufficient accuracy. Above, at least five decimal places are required, because the increasing number of atoms results in an unacceptable multiplication of many small mass errors. Beyond 1000 u, even six decimal places should be employed. However, the final results of these calculations may be reported with only four decimal places, because this is sufficient for most applications. If mass accuracies of significantly less than 1 mmu are to be expected, the use of six decimal places becomes necessary in any case. 

The limited resolution and mass accuracy of the early MALDI-TOF instruments made the combination of MALDI with magnetic sector instruments (Chap. 4.3) desirable, [148,149] but this set-up suffered from low shot-to-shot reproducibility and poor sensitivity getting a full scan spectrum required thousands of laser shots while scanning the magnet. Even though eutectic matrix mixtures were introduced to circumvent such problems, [90,91] the MALDI-magnetic sector combination never became established. 

Classically, high-resolution work is the domain of double-focusing magnetic sector instruments. More recently, TOP and to a certain degree triple quadrupole instruments are also capable of resolutions up to about 20,000. However, the rapid development of FT-ICR instruments has established those as the systems of choice if ultrahigh-resolution (>100,000) and highest mass accuracy (1 ppm) are required (Chap. 4.6). 

These techniques still are not sufficient for true specificity. As mass increases from 200 to 500, the number of formulas mathematically possible for that mass increases rapidly, and a mass accuracy yielding a single, unambiguous formula quickly becomes impossible (Kind and Fiehn, 2006 Kind et al., 2007). To reduce the possibilities, the atom types and abundance required to produce the observed isotope... 

The LTQ-Orbitrap has resolution and mass accuracy performance close to that of the LTQ-FTICR. As shown in Table 5.3 (column 4), LTQ-Orbitrap accurate mass measurements, using external calibration, for a set of 30 pharmaceutical compounds resulted in less than 2.3 ppm error. The data were acquired with a 4-min, 1-mL/min-flow-rate, positive-mode LC-ESI-MS method where all measurements were performed within 5h from mass calibration. Mass accuracies below 2-3 ppm, and often below 1 ppm, can be routinely achieved in both the positive- and negative-ion mode (Table 5.3, columns 4 and 5). The long-term mass stability of the LTQ-Orbitrap is not as consistent as observed for the LTQ-FTICR-MS, and the Orbitrap requires more frequent mass calibration however, mass calibration is a routine procedure that can be accomplished within 5-10 min. Figure 5.7 displays a 70-h (external calibration) mass accuracy plot for three negative ions collected with a LTQ-Orbitrap where the observed accuracy is 2.5 ppm or better with little mass drift for each ion. Overall, for routine accurate mass measurements on the Orbitrap, once-a-week calibration (for the desired polarity) is required however, considering the ease of the process, more frequent external calibration is not a burden. 

For chemical scenarios, the kinetic behavior of the reaction, the temperature and pressure increase rate must be known under runaway conditions in the interval between set pressure and maximum pressure. This implies a good knowledge of the thermo-chemical properties of the reaction mass. The required data are traditionally obtained from adiabatic calorimetric experiments [22, 25, 26]. Nevertheless, other calorimetric methods, especially dynamic DSC or Calvet experiments evaluated using the isoconversional approach, can also provide these data with accuracy and an excellent reliability for the temperature increase rate [27], as well as for the pressure increase [28, 29]. 

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