Mass accuracy

Mass accuracy describes correctness of the observed mass. It is defined as the ratio of mass error to the theoretical mass of an ion, typically expressed in parts per million (ppm). Mass accuracy and mass resolving power are equally important parameters when implementing MS in molecular identification because well-resolved spectral features are useful only if the mass is correct. However, mass resolving power is mainly determined by the mass analyzer, whereas mass accuracy is affected by both the mass analyzer and mass calibration. 

The mass accuracy is defined as the difference between measured accurate mass and calculated exact mass. The mass accuracy can be stated as absolute units of u (or mmu) or as relative mass accuracy in ppm, i.e., absolute mass accuracy divided by the mass it is determined for. As mass spectrometers tend to have similar absolute mass accuracies over a comparatively wide range, absolute mass accuracy represents a more meaningful way of stating mass accuracies than the more trendy use of ppm. 

Note Part per million (ppm) is simply a relative measure as are percent (%), permill (parts per thousand, %c), or parts per billion (ppb). 

Example A magnetic sector mass spectrometer allows for an absolute mass accuracy of 2-5 mmu in scanning mode over a range of about m/z 50-1500. At m/z 1200 an error of 3.5 mmu corresponds to inconspicuous 2.9 ppm, whereas the same error yields 70 ppm at m/z 50 which seems to be unacceptably large. 

Given infinite mass accuracy we should be able to identify the empirical formula of any ion from its exact mass. The emphasis is on infinite mass accuracy, however, and the meaning of infinite is perfectly illustrated by an example from McLafferty s book (Fig. 3.14). [13] Even though the compositions are restricted to CxHyNo 30o-4, there is a considerable number of isobaric ions of nominal mass 

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 absolute mass accuracy, Am/z, is defined as the difference between measured accurate mass and calculated exact mass  

Instead of stating the absolute mass accuracy in units of u, it can also be given as relative mass accuracy, dm/m, i.e., absolute mass accuracy divided by the mass it is determined for  

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TOF mass spectrometers are very robust and usable with a wide variety of ion sources and inlet systems. Having only simple electrostatic and no magnetic fields, their construction, maintenance, and calibration are usually straightforward. There is no upper theoretical mass limitation all ions can be made to proceed from source to detector. In practice, there is a mass limitation in that it becomes increasingly difficult to discriminate between times of arrival at the detector as the m/z value becomes large. This effect, coupled with the spread in arrival times for any one m/z value, means that discrimination between unit masses becomes difficult at about m/z 3000. At m/z 50,000, overlap of 50 mass units is more typical i.e., mass accuracy is no better than about 50-100 mass... 

Other techniques for mass measurement are available, but they are not as popular as those outlined above. These other methods include mass measurements on a standard substance to calibrate the instrument. The standard is then withdrawn, and the unknown is let into the instrument to obtain a new spectrum that is compared with that of the standard. It is assumed that there are no instrumental variations during this changeover. Generally, this technique is less reliable than when the standard and unknown are in the instrument together. Fourier-transform techniques are used with ion cyclotron mass spectrometers and give excellent mass accuracy at lower mass but not at higher. 

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. 

Easterling, M.L., Mize, T.H., and Amster, I. J., Routine part-per-million mass accuracy for high-mass ions space-charge effects in MALDI FT-ICR, Anal. Chem., 71, 624, 1999. 

Principles and Characteristics Mass spectrometry can provide the accurate mass determination in a direct measurement mode. For a properly calibrated mass spectrometer the mass accuracy should be expected to be good to at least 0.1 Da. Accurate mass measurements can be made at any resolution (resolution matters only when separating masses). For polymer/additive deformulation the nominal molecular weight of an analyte, as determined with an accuracy of 0.1 Da from the mass spectrum, is generally insufficient to characterise the sample, in view of the small mass differences in commercial additives. With the thousands of additives, it is obvious that the same nominal mass often corresponds to quite a number of possible additive types, e.g. NPG dibenzoate, Tinuvin 312, Uvistat 247, Flexricin P-1, isobutylpalmitate and fumaric acid for m = 312 Da see also Table 6.7 for m = 268 Da. Accurate mass measurements are most often made in El mode, since the sensitivity is high, and reference mass peaks are readily available (using various fluorinated reference materials). Accurate mass measurements can also be made in Cl... 

High resolution and mass accuracy (elemental composition)... 

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.

ToFs can also be used in combination with other mass analysers. Both hybrid sector and quadrupole systems are available. oaToF-MS has been interfaced to a quadrupole mass filter and hexapole gas collision cell, such as to allow recording of mass spectra and product ion spectra with good mass resolution (ca. 10000), high sensitivity, high mass range (ca. 10 000 Da) and high mass accuracy (<5ppm) [177,178]. QqToFMS may be fitted with API sources with flow-rates from nL... 

Fast screening techniques, such as temperature-resolved in-source filament pyrolysis and laser-assisted pyrolysis, benefit from the high cycle time and mass accuracy of FUCR-MS [214]. An additional advantage of FUCR-MS in the study of pyrolysis processes is that MS can be readily used for structural identification of desorption and pyrolysis products. 

Different options are available for LC-MS instruments. The vacuum system of a mass spectrometer typically will accept liquid flows in the range of 10-20 p,L min-1. For higher flow-rates it is necessary to modify the vacuum system (TSP interface), to remove the solvent before entry into the ion source (MB interface) or to split the effluent of the column (DLI interface). In the latter case only a small fraction (10-20 iLrnin ) of the total effluent is introduced into the ion source, where the mobile phase provides for chemical ionisation of the sample. The currently available commercial LC-MS systems (Table 7.48) differ widely in characteristics mass spectrometer (QMS, QQQ, QITMS, ToF-MS, B, B-QITMS, QToF-MS), mass range m/z 25000), resolution (up to 5000), mass accuracy (at best <5ppm), scan speed (up to 13000Das-1), interface (usually ESP/ISP and APCI, nanospray, PB, CF-FAB). There is no single LC-MS interface and ionisation mode that is readily suitable for all compounds... 

QToF-MS >20 000 ESI, APCI, MALDI Mass accuracy <5 ppm (m/z 150-900) resolution 5000 nanoflow... 

High mass accuracy (<5 ppm selectivity and elemental composition based on monoisotopic masses)... 

SEC-ESI-FTMS combines the size separation based technique of SEC with one of the most powerful mass spectrometric techniques of FTMS offering high mass accuracy (ppm), ultrahigh resolving power (>10(i) 6) and the capability to perform tandem mass spectrometry. The technique enables generation of oligomer elution profiles, which can be used for accurate calibration of standard SEC data. Coupling of SEC to ESI-MS is further described in ref. [710],... 

Lasers have advanced the analytical use of mass spectrometers to characterise additives in polymers, and routine application of MALDI is no longer limited to high molecular masses only. MALDI can now clearly produce isotopically resolved mass spectra of small molecules (<800 Da) in an L-ToF instrument, which can be used successfully for the characterisation of molecules of different chemical classes. High mass resolution with an improvement of mass accuracy to... 

The use of CIEF in combination with FTICR has been demonstrated in an analysis of the E. coli proteome (Jensen et al., 1999). For these experiments, E. coli was grown in a medium depleted of rare isotopes in order to increase the mass measurement accuracy. The high abundance isotopes are present at approximately 98.89% 12C, 99.63% 14N and 99.985% H. For peptides, the presence of rare isotopes does not significantly change the spectra but with undigested proteins, mass accuracy can be limited by the broadened distribution of ions of any given protein due to the incorporation... 

Currently PCR and mass spectrometry are performed by two separate instruments. However, there is no reason why PCR followed by simple automated cleanup and mass spectrometry cannot be incorporated into a single integrated instrument. Essentially every configuration of the modern ESI mass spectrometer has been used successfully for the analysis of PCR products, from the highest to the lowest resolution involving. Fourier transform ion cyclotron resonance (FTICR), triple quadrupole, quadrupole-time of flight (Q-TOF), and ion trap.22-24 MS discriminates between two structurally related PCR products by MW difference. Mass accuracy is needed to differentiate the... 

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