The factors and protocols that influence, accuracy, precision and uncertainty of accurate mass measurements by FT-ICR ms to validate the assignment of elemental composition

Abstract

The need for very accurate mass measurements of compounds is becoming more demanding with the expansion of the number of compounds in need of correct identification and with the limits of elemental analysis; a fast, robust analytical solution is sought. Fourier transform ion cyclotron resonance mass spectrometry (FT-ICR MS) is a very powerful tool for undertaking accurate mass measurements because of its high resolving power and mass accuracy. The use of FT-ICR MS in a high-throughput environment is limited due to the need of operator intervention required to obtain optimum accurate mass measurements. The lowest mass measurement errors (MME) are usually obtained using internal calibration, as the sample and calibrant ions inside the ICR cell experience the same conditions. However, internal calibration requires calibrant ions to be selected according to the mass of the sample ion, which requires operator intervention. External calibration is the preferred choice when performing accurate mass measurements in a high-throughput environment, as a calibration can be acquired independently of the sample. This study aims to demonstrate the use of ion population balancing using a dual sprayer approach to reduce MME. Population balancing between sample ions and calibrant ions can reduce the MME 40-fold. Ion populations across a chromatographic run can also vary greatly. The dual sprayer can also be employed to easily control the ion populations of both sample and calibrant entering the cell and can help to reduce ion suppression. The aim of the latter part of the work was to address the uncertainty of an accurate mass measurement performed using FT-ICR MS. In order to confidently select an elemental formula following an accurate mass measurement, a ‘cut-off’ limit or tolerance has to be selected. This tolerance can be set by calculating the uncertainty of the measurement; without an uncertainty component the measurement is meaningless. The work herein demonstrates how to calculate uncertainty of an accurate mass measurement with the aim that journals will adopt a procedure of only accepting an accurate mass measurement which quotes an uncertainty