Mass spectrometry with selective ion filtration by digital thresholding
View Patent ↗The methods described herein generally relate to characterization of large analytes, such as biomolecules, by molecular mass analysis. Specifically, the methods are directed to molecular mass analysis of singly— or multiply-charged ions by selective ion filtering carried out by a digital thresholding process.
1. A method of identifying a multiply-charged ion comprising:
a) specifying a first and a second digital signal thresholds on a ion identification apparatus, wherein the ion identification apparatus comprises:
i) an ion detector;
ii) a digitizer for converting an analog signal to a digital signal;
iii) an analog signal transfer means for transferring an analog signal from the detector to the digitizer; and
iv) a digital threshold filter, for setting a digital signal threshold, in digital data communication with the digitizer, in response to a digital signal input from the digitizer, independently outputting a digital signal to a data file only if the digital signal input is greater than the specified digital signal threshold;
wherein upon an ion identification apparatus measurement of the multiply-charged ion, the filtered digital signal output to the data file originates from the detection of the multiply-charged ion and excludes digital signal output from analog signals arising from singly-charged ions;
b) obtaining spectrum data of said multiply-charged ion at each of said first and second digital signal thresholds; and
c) subtracting said second spectrum data from said first spectrum data to identify said multiply-charged ion as a clean isotopically resolved spectrum.
2. The method of claim 1 wherein the multiply-charged ion is a biomolecule.
3. The method of claim 2 wherein the biomolecule is a nucleic acid, peptide, protein, lipid, or carbohydrate.
4. The method of claim 2 wherein the biomolecule comprises a non-covalently-bound small molecule.
5. The method of claim 1 wherein the singly-charged ions are biomolecule stabilizer additives.
6. The method of claim 5 wherein the stabilizer additives are one or more of: ampholytes, polyethylene glycol, glycerol, reducing agents, detergents or buffer salts, or any combination thereof.
7. The method of claim 1 wherein the analog signal is an analog voltage signal and the digital signal is a digital voltage signal.