Ribonucleotide and ribonucleoside determination by ambient pressure ion mobility spectrometry

Abstract

Detection limits and reduced mobilities for 12 ribonucleotides and 4 ribonucleosides were measured by ambient pressure electrospray ionization-ion mobility spectrometry (ESI-IMS). With the instrument used in this study it was possible to separate some of these compounds within mixtures. Detection limits reported for ribonucleotides and ribonucleosides ranged from 15 to 300 pmol and the reduced mobilities ranged from 41 to 56 suggesting that ambient pressure ESI-IMS may be used for their rapid and sensitive separation and detection. This report demonstrates that it was possible to use ion mobility spectrometry (IMS) to obtain a spectrum for the separation of nucleotides and nucleosides in less than 1 min. The application holds great promise for nucleotide analysis in the area of separating DNA fragments in genome sequencing and also for forensics DNA typing examinations used for the identification of blood stains in crime scenes and paternity testing.

Figure 1

Schematic cross-sectional side view of an ambient pressure ion mobility spectrometer.

Figure 2

ESI-IMS spectra for (a) adenosine, AMP, ADP, and ATP (b) cytidine, CMP, CDP and CTP (c) guanosine, GMP, GDP and GTP (d) uridine, UMP, UDP and UTP obtained in the negative ion mode. The reduced mobility values are indicated on the plots.

Figure 2

ESI-IMS spectra for (a) adenosine, AMP, ADP, and ATP (b) cytidine, CMP, CDP and CTP (c) guanosine, GMP, GDP and GTP (d) uridine, UMP, UDP and UTP obtained in the negative ion mode. The reduced mobility values are indicated on the plots.

Figure 2

ESI-IMS spectra for (a) adenosine, AMP, ADP, and ATP (b) cytidine, CMP, CDP and CTP (c) guanosine, GMP, GDP and GTP (d) uridine, UMP, UDP and UTP obtained in the negative ion mode. The reduced mobility values are indicated on the plots.

Figure 2

ESI-IMS spectra for (a) adenosine, AMP, ADP, and ATP (b) cytidine, CMP, CDP and CTP (c) guanosine, GMP, GDP and GTP (d) uridine, UMP, UDP and UTP obtained in the negative ion mode. The reduced mobility values are indicated on the plots.

Figure 3

ESI-IMS spectra for nucleotide mixtures. Spectrum (a) was a mixture of adenosine, AMP, ADP, and ATP. Spectrum (b) was a mixture of cytidine, CMP, CDP, and CTP. Spectrum (c) was a mixture of guanosine, GMP, GDP, and GTP whereas spectrum (d) was a mixture of uridine, UMP, UDP, and UTP. The numbers indicated on the plot were the reduced mobilities of the different ions observed.