To achieve the highest number of spectral averages per run, the flowrate of the analytical pump was decreased from 1.0 µL/min to 0.3 µL/min during elution of the nucleic acid to deliberately broaden the chromatographic peak. Despite some dilution during this peak parking, the lower flow rate enabled substantially more spectra to be collected in a single run without substantial detriment to either the desalting performance or observed MS signal, shown in Figure . This approach likewise increased the number of spectral averages recorded during MS acquisitions utilizing peak parking, 350 spectral averages at 120k resolution were recorded for tRNA-Phe over the roughly 2 minute chromatographic peak width, compared to only 125 spectral averages without peak parking. MS1 acquisitions of tRNA-Phe revealed multiple isoforms present within the sample. A portion of an MS1 spectrum shown in Figure (i.e., an expansion of a narrow section spanning m/z 870 to 895 from Figure ) showcases isotopically resolved species in the 28-charge state. Of the nine species observed with isotopic resolution, eight were putatively identified based on their intact mass and isotopic distributions. The two base peaks of m/z 890 and 878 corresponded to the tRNA gene sequence 25 and 3' adenosine loss, respectively. The other minor species were consistent with single nucleobase losses of guanine, methylguanine, or adenine (m/z 872, 884, 873) or additional methylations (m/z 879, 891). A low-abundance isoform of m/z 877 appeared to correspond to a guanine to adenine mutation. Theoretical isotopic fits based on these presumed identifications are shown in Figure , and their accompanying sequences in Table . The two most abundant tRNA-Phe isoforms of m/z 878 and 890 were subjected to targeted tandem MS analysis to assess their putative sequence assignments, as described in the following section.