Method of non-targeted complex sample analysis

Abstract

A method for non-targeted complex sample analysis which involves the following steps. A first step involves providing a database ( 16 ) containing identifying data of known molecules. A second step involves introducing a complex sample containing multiple unidentified molecules into a Fourier Transform Ion Cyclotron Mass Spectrometer ( 12 ) to obtain data regarding the molecules in the complex sample. A third step involves comparing the collected data regarding the molecules in the complex sample with the identifying data of known molecules in order to arrive at an identification through comparison of the molecules in the sample.

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       35 . A method for non-targeted analysis of a plurality of biological samples to identify one or more unidentified metabolite fragments of different intensities between samples, comprising the steps of: 
 a) introducing the plurality of biological samples, each of which contains a plurality of unidentified metabolites into a Fourier Transform Ion Cyclotron Resonance Mass Spectrometer (FTMS);    b) simultaneously obtaining identifying and quantifying data for the plurality of unidentified metabolite fragments detected in each of the biological samples introduced into the FTMS, wherein the identifying data comprises accurate mass and the quantifying data is intensity data;    c) creating a database comprising said identifying and quantifying data;    d) analyzing the database to determine metabolite fragments of different intensities between samples; and    e) identifying one or more metabolite fragments so determined using the identifying data by matching to a known database, or based upon their empirical formula or MS/MS fragment data.    
   
   
       36 . The method as defined in  claim 1 , wherein each of the biological samples is a biological extract of metabolites.  
   
   
       37 . The method as defined in  claim 1 , wherein the accurate mass is used to calculate the empirical formula of the metabolite fragments.  
   
   
       38 . The method as defined in  claim 3 , wherein the database is organized to permit searching for one or more known metabolite fragments by empirical formula.  
   
   
       39 . The method as defined in  claim 3 , wherein the database is organized to permit identification of unknown metabolite fragments by the empirical formulas of the metabolite fragments.  
   
   
       40 . The method as defined in  claim 1 , wherein the database is organized to permit searching for one or more known metabolite fragments by accurate mass.  
   
   
       41 . The method as defined in  claim 6 , wherein the database is organized to permit identification of unknown metabolite fragments by the accurate masses of the metabolite fragments.  
   
   
       42 . The method as defined in  claim 1 , wherein the database is organized to permit the comparison of one or more test samples to one or more control samples such that the intensity of metabolite fragments present in the test samples can be determined relative to the control samples and other test samples.  
   
   
       43 . The method as defined in  claim 1 , wherein the FTMS is used with a chromatographic separation system.  
   
   
       44 . The method as defined in  claim 1 , wherein the FTMS is equipped with a soft ionization source.  
   
   
       45 . The method as defined in  claim 1 , wherein the FTMS is equipped with an additional mass selective pre-separation system.  
   
   
       46 . The method as defined in  claim 1 , wherein the correlation of the data contained within the database from said biological samples are from a genetically modified test organism and a non genetically modified control organism with gene expression data from same said organisms for the purpose of determining the function of the genes affected by the genetic modification.  
   
   
       47 . The method as defined in  claim 1 , wherein the correlation of the data contained within the database from said biological samples are from an organism exposed to a test environment and a control environment with gene expression data from same said organism under same said conditions for the purpose of determining the function of the genes affected by the test environment.  
   
   
       48 . The method as defined in  claim 1 , wherein the correlation of the data contained within the database from said biological samples are from an organism at different stages of its development with gene expression data from same said organism under same said stages of its development for the purpose of determining the function of the genes affected by the changes in development of the organism.  
   
   
       49 . The method as defined in  claim 1 , wherein the database is organized to permit the comparison of any two or more samples to each other, such that the presence or absence of an intensity of metabolite fragments found in some samples but not in others is determined.  
   
   
       50 . The method as defined in  claim 1 , wherein the database is organized to permit the comparison of one or more test samples to one or more control samples such that the presence or absence of an intensity of metabolite fragments present in the test samples can be determined relative to the control samples or other test samples.  
   
   
       51 . A method for the non-targeted analysis of a plurality of biological samples to identify one or more unidentified metabolite fragments of different intensities between samples, comprising the steps of: 
 a) injecting the plurality of biological samples, each of which contains a plurality of unidentified metabolites into a Fourier Transform Ion Cyclotron Mass Spectrometer with or without the additional use of a chromatographic column;    b) ionizing the metabolites using a soft ionization source;    c) transferring the ionized metabolite fragments to an ion cyclotron resonance (ICR) cell with or without additional mass selective pre-separation;    d) separating and measuring said ions in the ICR cell with or without simultaneous MS/MS analysis occurring;    e) simultaneously determining accurate mass and intensity data of each of the ions detected;    f) transferring said data to a database that stores and organizes the data;    g) comparing biological samples contained within the database to one another to determine metabolite fragments of different intensities as between samples; and    h) identifying one or more metabolite fragments so determined using the identifying data by matching to a database of known metabolite fragments or based upon their empirical formula or MS/MS fragment data.    
   
   
       52 . A method for non-targeted analysis of a plurality of biological samples to identify one or more metabolite fragments from said samples when compared with a database of known metabolite fragments, comprising the steps of: 
 a) introducing the plurality of biological samples, each of which contains a plurality of unidentified metabolites into a Fourier Transform Ion Cyclotron Resonance Mass Spectrometer (FTMS);    b) simultaneously obtaining identifying and quantifying data for the plurality of unidentified metabolite fragments detected in each of the biological samples introduced into the FTMS, wherein the identifying data comprises accurate mass and the quantifying data is intensity data;    c) creating a database comprising said identifying and quantifying data;    d) comparing the database with a database of known metabolite fragments; and    e) identifying one or more metabolites so compared using the identifying data and matching said data to said database of known metabolite fragments.    
   
   
       53 . A method for non-targeted analysis of a plurality of biological samples to organize the database by fragment metabolic concentration, comprising the steps of: 
 a) introducing the plurality of biological samples, each of which contains a plurality of unidentified metabolites into a Fourier Transform Ion Cyclotron Resonance Mass Spectrometer (FTMS);    b) simultaneously obtaining identifying and quantifying data for the plurality of unidentified metabolite fragments detected in each of the biological samples introduced into the FTMS, wherein the identifying data comprises accurate mass and the quantifying data is intensity data;    c) creating a database comprising said identifying and quantifying data; and    d) organizing the database by concentration of the metabolite fragments.    
   
   
       54 . A method for non-targeted analysis of a plurality of biological samples, comprising the steps of: 
 a) introducing the plurality of biological samples, each of which contains a plurality of unidentified metabolites into a Fourier Transform Ion Cyclotron Resonance Mass Spectrometer (FTMS);    b) simultaneously obtaining, identifying and quantifying data for the plurality of unidentified metabolite fragments detected in each of the biological samples introduced into the FTMS, wherein the identifying data comprises accurate mass and the quantifying data is intensity data;    c) creating a database comprising said identifying and quantifying data;    d) analyzing the database, wherein the analyzing step is selected from the group consisting of: analyzing the database to determine metabolite fragments of different intensities between samples and identifying one or more metabolite fragments so determined using the identifying data by matching to a database of known metabolite fragments, or based upon their empirical formula or MS/MS fragment data; comparing the database with a database of known metabolite fragments and identifying one or more metabolite fragments so compared using the identifying data and matching said data to said database of known metabolite fragments; and organizing the database by concentration of metabolite fragments.