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US 10,591,448 B2Granted
Structural elucidation of isotopically labeled analytes
AssigneeWaters Technologies Corporation
InventorGiuseppe Astarita
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Abstract
The present disclosure relates to structural elucidation of isotopically labeled analytes using chromatography-MS and data-independent acquisition. The present disclosure can be used for rapid screening of analytes, such as the metabolome, including the simultaneous collection of both qualitative and quantitative information of known and unknown analytes.
Claims13
A method of analyzing a biological sample, the method comprising:
- (a) isotopically labeling metabolites in the biological sample;
- (b) separating the isotopically labeled metabolites using chromatography;
- (c) generating a low energy fragmentation mass spectrum of parent ions of the isotopically labeled metabolites and generating a high energy fragmentation mass spectrum of fragment ions of the parent ions, each mass spectrum generated using a mass spectrometer operating in data independent mode;
- (d) identifying at least one characteristic set of peaks in the low energy fragmentation mass spectrum corresponding to the parent ions of the isotopically labeled metabolites;
- (e) identifying at least one characteristic set of peaks in the high energy fragmentation mass spectrum that are substantially similar to the characteristic set of peaks in the low energy fragmentation mass spectrum and correspond to the fragment ions of the parent ions;
- and
- (f) determining a chemical structure of the ions of the isotopically labeled metabolites from the combination of at least one characteristic set of peaks in the high fragmentation mass corresponding to the fragment ions of the parent ions and the at least one characteristic set of peaks in the low energy fragmentation mass corresponding to the parent ions of the isotopically labeled metabolites.
4 more elements
A method of analyzing a biological sample, the method comprising:
- (a) isotopically labeling metabolites in the biological sample;
- (b) separating the isotopically labeled metabolites using chromatography;
- (c) providing an ion source for generating ions of the isotopically labeled metabolites;
- (d) passing said ions of the isotopically labeled metabolites to a fragmentation means including a collision cell;
- (e) operating said fragmentation means in a high energy mode wherein at least a portion of said ions of the isotopically labeled metabolites are fragmented to produce daughter ions;
- (f) generating a mass spectrum of ions emerging from the fragmentation means operating in the high energy mode as a high energy fragmentation mass spectrum, the mass spectrum generated using a mass spectrometer operating in data independent mode;
- (g) switching the fragmentation means to operate in a low energy mode wherein substantially fewer ions of the isotopically labeled metabolites are fragmented;
- (h) generating a mass spectrum of ions emerging from the fragmentation means operating in the low energy mode as a low energy fragmentation mass spectrum;
- (i) repeating steps (e)-(h) a plurality of times;
- (j) identifying at least one characteristic set of peaks in the low energy fragmentation mass spectrum, the mass spectrum generated using a mass spectrometer operating in data independent mode;
- (k) identifying at least one characteristic set of peaks in the high energy fragmentation mass spectrum that is substantially similar to the characteristic set of peaks in the low energy fragmentation mass spectrum;
- (l) determining a chemical structure associated with the at least one characteristic set of peaks in the high energy fragmentation mass spectrum and the at least one characteristic set of peaks in the low energy fragmentation mass spectrum.
9 more elements
Dependencies are inferred from the claim text.