Factors determining the performance of triple quadrupole, quadrupole ion trap and sector field mass spectrometers in electrospray ionization tandem mass spectrometry of oligonucleotides. 1. Comparison of performance characteristics
Article first published online: 31 MAY 2001
Copyright © 2001 John Wiley & Sons, Ltd.
Rapid Communications in Mass Spectrometry
Volume 15, Issue 13, pages 1045–1052, 15 July 2001
How to Cite
Premstaller, A., Ongania, K.-H., Huber, C. G. (2001), Factors determining the performance of triple quadrupole, quadrupole ion trap and sector field mass spectrometers in electrospray ionization tandem mass spectrometry of oligonucleotides. 1. Comparison of performance characteristics. Rapid Commun. Mass Spectrom., 15: 1045–1052. doi: 10.1002/rcm.336
- Issue published online: 31 MAY 2001
- Article first published online: 31 MAY 2001
- Manuscript Accepted: 2 MAY 2001
- Manuscript Received: 30 APR 2001
- Austrian Science Fund. Grant Number: P13442-PHY
The performance of triple-stage quadrupole (TSQ), quadrupole ion trap (QIT), and double focusing sector field (DFSF) mass spectrometers for the generation of fragment ions to obtain sequence information about oligonucleotides was compared. Upon electrospray ionization (ESI), the charge-state distribution of candidate precursor ions not only varied significantly with the type of mass spectrometer, but also with the size and sequence of the investigated oligonucleotides. While concentration limits of detection for an octanucleotide were in the 100 pmol/L range on the QIT and in the 5–10 nmol/L range on the TSQ and DFSF instruments, those of a 24-mer were in the 2–13 nmol/L range on all three instruments. Reproducibility of mass determination, an important prerequisite for reliable identification of fragment ions, was highest on the TSQ with 0.0037% relative standard deviation over three days. Finally, the tandem mass spectra of a dimethoxytritylated pentanucleotide recorded on the three instruments were compared. Relatively simple spectra dominated by complete series of fragment ions of the (a-B) and w type were obtained on the QIT. Complete series of (a-B) and w ions were also observed on the TSQ. However, additional fragments belonging to the b, c, d, x and z series were found in the spectrum. In the spectrum recorded after in-source fragmentation in the DFSF, only fragments corresponding to the loss of a nucleobase and a complete series of w ions were observed. All three mass spectrometers were suitable for the generation of fragment ions, from which the complete nucleotide sequence of the pentanucleotide could be deduced. Copyright © 2001 John Wiley & Sons, Ltd.