Systems and methods for identifying compounds from MS/MS data without precursor ion information
Systems and methods are provided for identifying a precursor ion without using any a priori precursor ion information. In one method, a sample is analyzed using a tandem mass spectrometer, producing at least one measured product ion spectrum from a precursor mass-to-charge ratio range. The at least one measured product ion spectrum are received. A subset of measured product ions is selected from the at least one measured product ion spectrum. A list of candidate compounds is created by searching a dictionary of potential compounds that includes one or more predicted product ions for each of the potential compounds using the subset of measured product ions. A candidate compound on the list is selected as the identified compound. In another method, the measured product ions are assumed to correspond to shortened forms of the peptide and a protein database is searched for shortened forms of the peptide.
1. A system for identifying a compound from tandem mass spectrometry product ions without using any a priori precursor ion information, comprising:
a tandem mass spectrometer that analyzes a sample producing at least one measured product ion spectrum from a precursor mass-to-charge ratio range; and
a processor in communication with the tandem mass spectrometer that
(a) receives the at least one measured product ion spectrum from the tandem mass spectrometer,
(b) selects a measured product ion from the at least one measured product ion spectrum,
(c) creates a list of candidate compounds by searching a dictionary of potential compounds with a mass of the measured product ion, wherein the dictionary includes one or more predicted product ions for each of the potential compounds and wherein the dictionary is searched using a key that is a product ion mass to retrieve a list of candidate compounds that could produce the product ion mass,
(d) selects a next measured product ion from the at least one measured product ion spectrum,
(e) creates a next list of candidate compounds by searching the dictionary of potential compounds with a mass of the next measured product ion,
(f) examines all lists of candidate compounds created for candidate compounds common to all the lists, and
(g) repeats steps (d)-(f) until one candidate compound is common to all the lists and identifies the one candidate compound as the identified compound of the product ions selected.
2. The system of claim 1 , wherein the measured product ion spectrum comprises peptide fragments, the dictionary of potential compounds comprises predicted peptides for proteins, the one or more predicted product ions for each of the potential compounds comprise one or more fragments of peptides, the list of candidate compounds comprises candidate peptides, and the identified compound comprises an identified peptide.
3. The system of claim 1 , wherein the potential compounds comprise one or more of sugars, lipids, DNA, and RNA.
4. The system of claim 1 , wherein the processor selects measured product ions from the at least one measured product ion spectrum by selecting measured mono-isotopic product ions with the highest intensity from the at least one measured product ion spectrum.
5. The system of claim 1 , wherein the processor selects a subset of measured product ions by selecting a number of measured product ions from the at least one measured product ion spectrum with a mass-to-charge ratio (m/z) greater than a threshold value.
6. The system of claim 1 , wherein the tandem mass spectrometer analyzes a sample by
dividing a mass range into two or more precursor mass windows, and
fragmenting each precursor mass window of the two or more mass windows, producing a measured product ion spectrum for each precursor mass window.
7. The system of claim 1 , wherein the processor further compares the identified compound to the range and
if the identified compound mass-to-charge ratio (m/z) is not found in the precursor mass-to-charge ratio range, determines that the identified compound is a modified form.
8. The system of claim 1 , wherein the processor further analyzes a list of modifications to determine a modification that puts the mass-to-charge ratio of the modified identified compound in the range.
9. The system of claim 1 , wherein the processor further subtracts measured product ions of the identified compound from the measured product ion spectrum and executes steps (b)-(g) again to identify another compound in the sample.
10. A method for identifying a compound from tandem mass spectrometry product ions without using any a priori precursor ion information, comprising:
(a) analyzing a sample using a tandem mass spectrometer, producing at least one measured product ion spectrum from a precursor mass-to-charge ratio range;
(b) receiving the at least one measured product ion spectrum from the tandem mass spectrometer using a processor;
(c) selecting a measured product ion from the at least one measured product ion spectrum using the processor;
(d) creating a list of candidate compounds by searching a dictionary of potential compounds with a mass of the measured product ion, wherein the dictionary includes one or more predicted product ions for each of the potential compounds and wherein the dictionary is searched using a key that is a product ion mass to retrieve a list of candidate compounds that could produce the product ion mass using the processor;
(e) selecting a next measured product ion from the at least one measured product ion spectrum using the processor;
(f) creating a next list of candidate compounds by searching the dictionary of potential compounds with a mass of the next measured product ion using the processor;
(g) examining all lists of candidate compounds created for candidate compounds common to all the lists using the processor; and
(h) repeating steps (e)-(g) until one candidate compound is common to all the lists and identifies the one candidate compound as the identified compound of the product ions selected using the processor.
11. The method of claim 10 , wherein selecting measured product ions comprises selecting a number of measured mono-isotopic product ions with the highest intensity from the at least one measured product ion spectrum.
12. The method of claim 10 , comprising selecting a number of measured product ions from the at least one measured product ion spectrum with a mass-to-charge ratio (m/z) greater than a threshold value.
13. A computer program product, comprising a non-transitory and tangible computer-readable storage medium whose contents include a program with instructions being executed on a processor so as to perform a method for identifying a precursor ion from tandem mass spectrometry product ions without using any a priori precursor ion information, the method comprising:
(a) providing a system, wherein the system comprises one or more distinct software modules, and wherein the distinct software modules comprise a measurement module and an identification module;
(b) receiving from a tandem mass spectrometer at least one measured product ion spectrum from a precursor mass-to-charge ratio range produced by analyzing a sample using the measurement module;
(c) selecting a measured product ion from the at least one measured product ion spectrum using the identification module;
(d) creating a list of candidate compounds by searching a dictionary of potential compounds with a mass of the measured product ion, wherein the dictionary includes one or more predicted product ions for each of the potential compounds and wherein the dictionary is searched using a key that is a product ion mass to retrieve a list of candidate compounds that could produce the product ion mass using the identification module;
(e) selecting a next measured product ion from the at least one measured product ion spectrum using the identification module;
(f) creating a next list of candidate compounds by searching the dictionary of potential compounds with a mass of the next measured product ion using the identification module;
(g) examining all lists of candidate compounds created for candidate compounds common to all the lists using the identification module; and
(h) repeating steps (e)-(g) until one candidate compound is common to all the lists and identifies the one candidate compound as the identified compound of the product ions selected using the identification module.
14. The computer program product of claim 13 , wherein selecting measured product ions comprises selecting a number of measured mono-isotopic product ions with the highest intensity from the at least one measured product ion spectrum.
15. The computer program product of claim 13 , comprising selecting a number of measured product ions from the at least one measured product ion spectrum with a mass-to-charge ratio (m/z) greater than a threshold value.