Quantitation Method Using Isotope Labeled Internal Standard Substance, Analysis System for Executing the Quantitation Method, and Program for the Analysis

Abstract

Problem: An object of the present invention is to quantitate with good accuracy, furthermore, quantitate absolutely, one or a plurality of biological molecules in a sample such as a tissue, a biological fluid, a cell, a cell organ or protein complex. Solution: By adding a metabolically isotope labeled biological molecule as an internal standard substance and measuring with a mass spectrometer, quantitating with good accuracy one or a plurality of target molecules in a sample has become possible. In addition, by performing waveform separation processing during mass analysis, a highly accurate quantitative analysis method of mass analysis is provided.

1 - 20 . (canceled) 
     
     
         21 . An internal standard substance that is a metabolically isotope labeled biological molecule to be used in the method according to any one of claims  1  to  20 . 
     
     
         22 . A cell containing an internal standard substance that is a metabolically isotope labeled biological molecule to be used in the method according to any one of claims  1  to  20 . 
     
     
         23 . A reagent for use in the method according to any one of claims  1  to  20 , comprising an internal standard substance that is a metabolically isotope labeled biological molecule. 
     
     
         24 . A reagent for use in the method according to any one of claims  1  to  20 , comprising a cell containing an internal standard substance that is a metabolically isotope labeled biological molecule. 
     
     
         25 - 26 . (canceled) 
     
     
         27 . A program for causing a computer receiving data on mass analysis of one or a plurality of biological molecules in a sample obtained in a mass spectroscopic device to quantitate one or a plurality of biological molecules in the sample, the program comprising performing the steps of:
 (1) receiving an analytical result obtained by analyzing with the mass spectroscopic device each biological molecule in the each sample, wherein an internal standard substance that is a metabolically isotope labeled biological molecule or a cell containing an internal standard substance that is a metabolically isotope labeled biological molecule is added to the sample;   (2) identifying the biological molecule from the analytical result of mass analysis; and   (3) determining a ratio of intensities between a labeled peak and an unlabeled peak of each biological molecule to quantitate the biological molecule.   
     
     
         28 . A program for causing a computer receiving data on the mass analysis of one or a plurality of biological molecules in a sample obtained in a mass spectroscopic device to quantitate the one or a plurality of biological molecules in the sample, the program comprising performing the steps of:
 (1) receiving an analytical result obtained by analyzing with the mass spectroscopic device each biological molecule in each sample, wherein an internal standard substance that is a metabolically isotope labeled biological molecule, or a cell containing an internal standard substance that is a metabolically isotope labeled biological molecule is added to the sample, and a biological molecule is extracted and fractionated from the each sample;   (2) identifying the biological molecule from the analytical result of mass analysis; and   (3) determining a ratio of intensities between a labeled peak and an unlabeled peak of each biological molecule to quantitate said biological molecule.   
     
     
         29 . The program according to  claim 27  or  28 , wherein analysis for a plurality of samples is performed, and further comprising performing of comparing a peak intensity ratio of each biological molecule between each sample. 
     
     
         30 . The program according to any one of  claims 27  to  29 , which is a program for quantitating absolutely one or a plurality of biological molecules in the sample, and wherein the biological molecule in the internal standard substance that is the metabolically isotope labeled biological molecule, or in the cell containing the internal standard substance that is the metabolically isotope labeled biological molecule, is present in a known quantity. 
     
     
         31 . An analysis system for analyzing data on mass analysis of a first biological molecule and a second biological molecule that has been isotope labeled with for corresponding to the first biological molecule, comprising:
 a mass spectroscopic device for performing mass analysis of the first and the second biological molecules; and   a control unit comprising a data input and output unit for receiving the data on the mass analysis of the biological molecule from the mass spectroscopic device,   wherein the control unit comprises:   a processing unit for determining a first m/z value of the first biological molecule for which a sequence of a biological molecular component has been determined, and a second m/z value of the second biological molecule for which a sequence of a biological molecular component has been determined, and, extracting data containing a mass spectrum against m/z values and a mass chromatogram of the first biological molecule and the second biological molecule from data on mass analysis obtained in the mass spectroscopic device, based on the first and the second m/z values; and   a waveform processing unit for receiving the mass chromatogram at the first and the second m/z values from the processing unit, and performing waveform separation processing of the mass chromatogram against a time axis.   
     
     
         32 . The analysis system according to  claim 31 , wherein the determination of the first and the second m/z values is carried out in such a way that the determination of the second m/z value is made by determining from the first m/z value of the first biological molecule for which the sequence of the biological molecular component has been determined, based on information on the number of non-isotope labeled component of the first biological molecule and an electric charge of the first biological molecule, or the determination of the first m/z value is made by determining from the second m/z value of the second biological molecule for which the sequence of the biological molecular component has been determined, based on information on the number of isotope labeled components of the second biological molecule and an electric charge of the second biological molecule. 
     
     
         33 . The analysis system according to  claim 31  or  32 , wherein the control unit further comprises a computation unit for calculating an area of a waveform derived from an individual biological molecule obtained by waveform separation processing from the waveform separation processing unit, and calculating a ratio (an area of a first waveform)/(an area of a second waveform) from a value of the area of the first waveform derived from an individual biological molecule at the first m/z value and a value of the area of the second waveform derived from an individual biological molecule at the second m/z value. 
     
     
         34 . The analysis system according to any one of  claims 31  to  33 , wherein the control unit further comprises an information addition unit for performing, after waveform separation, classification for each sequence of a component of the biological molecule, retrieving the sequence with a database containing information on the function of biological molecules, and linking the sequence to function from the result of the verification so as to group. 
     
     
         35 . The analysis system according to any one of  claims 31  to  34 , wherein the control unit further comprises a computation unit for identifying the biological molecule from the sequence of the biological molecular component, and determining, from a quantitative value of each biological molecular component constituting the biological molecule, a mean value to serve as a representative quantitative value for the biological molecule. 
     
     
         36 . The analysis system according to any one of  claims 31  to  35 , wherein the control unit further comprises an information addition unit for retrieving the sequence of the biological molecular component with the database containing information on the function of biological molecules, and acquiring information on function of the biological molecule. 
     
     
         37 . The analysis system according to any one of  claims 31  to  36 , wherein when mass analyses of the first and the second biological molecules are performed in the presence of an internal standard substance,
 the waveform processing unit for receiving a mass chromatogram at the m/z value of the internal standard substance and further performing waveform separation processing to obtain a waveform specific to the internal standard substance, and 
 the computation unit for calculating an area of the waveform specific to the internal standard substance from the waveform processing unit and at the same time further calculating a ratio (the area of the first waveform)/(the area of the waveform specific to the internal standard substance), and/or a ratio (the area of the second waveform)/(the area of the waveform specific to the internal standard substance). 
 
     
     
         38 . An analysis system for analyzing data on mass analysis of an internal standard substance, which is an isotope labeled biological molecule for corresponding to a first and a second biological molecule, comprising:
 a mass spectroscopic device for performing mass analysis of the first and the second biological molecules as well as the isotope labeled biological molecule; and   a control unit comprising a data input and output unit for receiving the data on mass analysis of the biological molecules from the mass spectroscopic device;   wherein the control unit comprises:   a processing unit for determining a first m/z value and a second m/z value of the biological molecule for which a sequence of a biological molecular component has been determined and a m/z value of an internal standard substance which is isotope labeled biological molecule for which a sequence of a biological molecular component has been determined, and, extracting data containing mass spectrum against the m/z value and a mass chromatogram of the first and the second biological molecules as well as of the biological molecule constituting the internal standard substance from the data on mass analysis obtained in the mass spectroscopic device, based on the m/z values of the first and the second as well as the internal standard substance; and   a waveform processing unit for receiving the mass chromatogram at the m/z values of the first and the second biological molecules as well as the internal standard substance from the processing unit and performing waveform separation processing of the mass chromatogram.   
     
     
         39 . The analysis system according to  claim 38 , wherein the determination of the first m/z value and the second m/z value, as well as the m/z value of the internal standard substance is carried out in such a way that the determination of the m/z value of the biological molecule constituting the internal standard substance is made by determining, from the first m/z value and the second m/z value of the first and the second biological molecules for which a sequence of the biological molecular component has been determined, based on information on the number of non-isotope labeled components of the first and the second biological molecules and an electric charge of the first and the second biological molecules, or the determination of the first m/z value and the second m/z value of the first and the second biological molecules is made by determining, from the m/z value of the internal standard substance for which a sequence of the biological molecular component has been determined, based on information on the number of isotope labeled biological molecular components of the internal standard substance and an electric charge of the biological molecule constituting the internal standard substance. 
     
     
         40 . The analysis system according to  claim 38 , wherein the control unit further comprises a computation unit for calculating an area of a waveform derived from an individual biological molecule obtained by waveform separation processing from the waveform separation processing unit, and calculating a ratio (an area of a first waveform)/(an area of a waveform specific to the internal standard substance), and/or, a ratio (an area of a second waveform)/(an area of a waveform specific to the internal standard substance) from values of the areas of the first waveform and the second waveform derived from specific biological molecules at the first and the second m/z values, and a value of the area of the internal standard substance waveform derived from an individual biological molecule at the m/z value of the internal standard substance. 
     
     
         41 . The analysis system according to  claim 38 , wherein the control unit further comprises an information addition unit for performing, after waveform separation, classification for each sequence of the biological molecular component, retrieving the sequence with a database containing information on the function of biological molecules, and linking the sequence with function from the result of the verification so as to group. 
     
     
         42 . The analysis system according to  claim 38 , wherein the control unit further comprises a computation unit for identifying the biological molecule from the sequence of the biological molecular component, and determining, from a quantitative value of each biological molecular component constituting the biological molecule, a mean value to serve as a representative quantitative value for the biological molecule. 
     
     
         43 . The analysis system according to  claim 38 , wherein the control unit further comprises an information addition unit for retrieving the sequence of the biological molecular component with the database containing information on the function of biological molecules, and acquiring information on function of the biological molecule. 
     
     
         44 . The analysis system according to  claim 38 , wherein the isotope used in the isotope labeling is any isotope selected from the group consisting of  2 H,  13 C,  15 N,  17 O,  18 O,  33 P and  34 S, as well as combinations thereof. 
     
     
         45 . The analysis system according to  claim 38 , wherein the determination of the sequence of the component is carried out by MS/MS processing. 
     
     
         46 . The analysis system according to  claim 45 , wherein the determination of the sequence of the component is carried out by retrieving data obtained by MS/MS processing with a database regarding a biological molecule. 
     
     
         47 . The analysis system according to  claim 38 , wherein the biological molecule is a biological molecule that has been separated from the sample over time. 
     
     
         48 . The analysis system according to  claim 38 , wherein the biological molecule is any molecule selected from the group consisting of a protein, a lipid, a sugar chain and a nucleic acid, as well as combinations thereof. 
     
     
         49 . An analysis method for analyzing data on mass analysis of a first biological molecule and a second biological molecule that has been isotope labeled for corresponding to the first biological molecule, comprising the steps of:
 determining, among the data on the mass analysis of the biological molecule measured in a mass spectroscopic device, a first m/z value of the first biological molecule for which a sequence of a biological molecular component has been determined, and a second m/z value of the second biological molecule for which a sequence of a biological molecular component has been determined,   extracting data containing a mass spectrum against m/z values and a mass chromatogram of the first biological molecule and the second biological molecule from the data on the mass analysis, based on the first and the second m/z values; and   performing waveform separation processing of the mass chromatogram of the first and the second biological molecules.   
     
     
         50 . The analysis method according to  claim 49 , wherein the determination of the first m/z value and the second m/z value is carried out in such a way that the determination of the second m/z value is made by determining from the first m/z value of the first biological molecule for which the sequence of the biological molecular component has been determined, based on information on the number of non-isotope labeled components of the first biological molecule and an electric charge of the first biological molecule, or the determination of the first m/z value of the second biological molecule is made by determining from the second m/z value of the second biological molecule for which the sequence of the biological molecular component has been determined, based on information on the number of isotope labeled components of the second biological molecule and an electric charge of the second biological molecule. 
     
     
         51 . The analysis method according to  claim 49  or  claim 50 , further comprising the steps of:
 calculating an area of a waveform derived from an individual biological molecule obtained by the waveform separation processing step; and 
 calculating a ratio (an area of a first waveform)/(an area of a second waveform) from a value of the area of the first waveform derived from an individual biological molecule at the first m/z value, and a value of the area of the second waveform derived from an individual biological molecule at the second m/z value. 
 
     
     
         52 . The analysis method according to any one of  claims 49  to  51 , further comprising the steps of:
 performing, after the waveform separation, classification for each sequence of a component of the biological molecule, 
 retrieving the sequence with a database containing information on the function of biological molecules, and 
 linking the sequence to function from the result of the verification so as to group. 
 
     
     
         53 . The analysis method according to any one of  claims 49  to  52 , further comprising the steps of:
 identifying the biological molecule from the sequence of the biological molecular component; and 
 determining, from a quantitative value of each biological molecular component constituting the biological molecule, a mean value to serve as a representative quantitative value for the biological molecule. 
 
     
     
         54 . The analysis method according to any one of  claims 49  to  53 , further comprising retrieving the sequence of the biological molecular component with a database containing information on the function of biological molecules so as to acquire information on the function of the biological molecule. 
     
     
         55 . The analysis method according to any one of  claims 49  to  54 , further comprising the steps of:
 performing determination of a m/z value of the internal standard substance along with m/z values of the first and the second biological molecules, extraction of data containing a mass chromatogram of the internal standard substance, waveform separation processing of the mass chromatogram of the internal standard substance and calculation of an area of a waveform specific to the internal standard substance; and 
 calculating a ratio (the area of the first waveform)/(the area of the waveform specific to the internal standard substance), and/or, a ratio (the area of the second waveform)/(the area of the waveform specific to the internal standard substance). 
 
     
     
         56 . An analysis method for analyzing data on mass analysis of an internal standard substance, which is a biological molecule that has been isotope labeled for corresponding to a first and a second biological molecule, comprising the steps of:
 determining, among the data on the mass analysis of the biological molecules measured in a mass spectroscopic device, a first m/z value and a second m/z value of the first and the second biological molecules for which a sequence of a biological molecular component has been determined, and a m/z value of the biological molecule constituting the internal standard substance for which a sequence of a biological molecular component has been determined;   extracting data containing a mass spectrum against m/z values and a mass chromatogram of the first and the second biological molecules as well as of the biological molecule constituting the internal standard substance from the data on the mass analysis, based on the m/z values of the first and the second as well as of the internal standard substance; and   performing waveform separation processing on the mass chromatogram of the first and second of the biological molecules, as well as the biological molecule constituting the internal standard substance.   
     
     
         57 . The analysis method according to  claim 56 , wherein the determination of the first m/z value and the second m/z value, as well as the m/z value of the internal standard substance is carried out in such was that the determination of the m/z value of the biological molecule constituting the internal standard substance is made by determining from the first m/z value and the second m/z value of the first and the second biological molecules for which a sequence of the biological molecular component has been determined, based on information on the number of non-isotope labeled components of the first and the second biological molecules and an electric charge of the first and the second biological molecules, or the determination of the first m/z value and the second m/z value is made by determining, from the m/z value of the internal standard substance for which a sequence of the biological molecular component has been determined, based on information on the number of isotope labeled biological molecular components of the internal standard substance and an electric charge of the biological molecule constituting the internal standard substance. 
     
     
         58 . The analysis method according to  claim 56  or  57 , further comprising the steps of:
 calculating an area of a waveform derived from an individual biological molecule obtained by the waveform separation processing step; and 
 performing calculation a ratio (an area of a first waveform)/(an area of a waveform specific to the internal standard substance), and/or, a ratio (an area of a second waveform)/(an area of a waveform specific to the internal standard substance) from values of the areas of the first and the second waveforms derived from specific biological molecules at the first and the second m/z values, and a value of the area of the internal standard substance waveform derived from an individual biological molecule at m/z value of the internal standard substance. 
 
     
     
         59 . The analysis method according to any one of  claims 56  to  58 , further comprising the steps of:
 performing, after the waveform separation, classification for each sequence of the component of the biological molecule; 
 retrieving the sequence with a database containing information on the function of biological molecules; and 
 linking the sequence to function from the result of the verification so as to group. 
 
     
     
         60 . The analysis method according to any one of  claims 56  to  59 , further comprising the steps of:
 identifying the biological molecule from the sequence of the biological molecular component; and 
 determining, from a quantitative value of each biological molecular component constituting the biological molecule, a mean value to serve as a representative quantitative value for the biological molecule. 
 
     
     
         61 . The analysis method according to any one of  claims 56  to  60 , further comprising retrieving the sequence of the biological molecular component with a database containing information on the function of biological molecules so as to acquire information on the function of the biological molecule. 
     
     
         62 . The analysis method according to any one of  claims 49  to  61 , wherein the isotope used in the isotope labeling is any isotope selected from the group consisting of  2 H,  13 C,  15 N,  17 O,  18 O,  33 P and  34 S, as well as combinations thereof. 
     
     
         63 . The analysis method according to any one of  claims 49  to  62 , wherein the determination of the sequence of the component is carried out by MS/MS processing. 
     
     
         64 . The analysis method according to  claim 63 , wherein the determination of the sequence of the component is carried out by retrieving data obtained by MS/MS processing with a database regarding a biological molecule. 
     
     
         65 . The analysis method according to any one of  claims 49  to  64 , wherein the biological molecule is a biological molecule that has been separated from the sample over time. 
     
     
         66 . The analysis method according to any one of  claims 49  to  65 , wherein the biological molecule is any molecule selected from the group consisting of a protein, a lipid, a sugar chain and a nucleic acid, as well as combinations thereof. 
     
     
         67 . A program for causing a computer receiving data obtained in a mass spectroscopic device to analyze the data in which the data is related to mass analysis of a first biological molecule and a second biological molecule, which has been isotope labeled for corresponding to the first biological molecule, the program performing the steps of:
 determining, among the data on the mass analysis of the biological molecule measured in the mass spectroscopic device, a first m/z value of the first biological molecule for which a sequence of a biological molecular component has been determined, and a second m/z value of the second biological molecule for which a sequence of a biological molecular component has been determined,   extracting data containing a mass spectrum against m/z values and a mass chromatogram of the first biological molecule and the second biological molecule from the data on the mass analysis, based on the first and the second m/z values; and   performing waveform separation processing of the mass chromatogram of the first and the second biological molecules.   
     
     
         68 . The program according to  claim 67 , wherein the determination of the first m/z value and the second m/z value is carried out in such a way that the determination of the second m/z value is made by determining from the first m/z value of the first biological molecule for which the sequence of the biological molecular component has been determined, based on information on the number of non-isotope labeled components of the first biological molecule and an electric charge of the first biological molecule, or the determination of the first m/z value of the second biological molecule is made by determining from the second m/z value of the second biological molecule for which the sequence of the biological molecular component has been determined, based on information on the number of isotope labeled components of the second biological molecule and an electric charge of the second biological molecule. 
     
     
         69 . The program according to  claim 67  or  68 , further comprising performing the steps of:
 calculating an area of a waveform derived from an individual biological molecule obtained by the waveform separation processing step; and 
 calculating a ratio (an area of a first waveform)/(an area of a second waveform) from a value of the area of the first waveform derived from an individual biological molecule at the first m/z value, and a value of the area of the second waveform derived from an individual biological molecule at the second m/z value. 
 
     
     
         70 . The program according to any one of  claims 67  to  69 , further comprising performing the steps of:
 performing, after the waveform separation, classification for each sequence of a component of the biological molecule, 
 retrieving the sequence with a database containing information on the function of biological molecules, and 
 linking the sequence to function from the result of the verification so as to group. 
 
     
     
         71 . The program according to any one of  claims 67  to  70 , further comprising performing the steps of:
 identifying the biological molecule from the sequence of the biological molecular component; and 
 determining, from a quantitative value of each biological molecular component constituting the biological molecule, a mean value to serve as a representative quantitative value for the biological molecule. 
 
     
     
         72 . The program according to any one of  claims 67  to  71 , further comprising performing of retrieving the sequence of the biological molecular component with a database containing information on the function of biological molecules so as to acquire information on the function of the biological molecule. 
     
     
         73 . The program according to any one of  claims 67  to  72 , further comprising performing the steps of:
 obtaining, in the mass spectroscopic device, data on the mass analysis of an internal standard substance along with the mass analysis of the first and the second biological molecules, 
 extracting a mass chromatogram of the internal standard substance from the data on the mass analysis, and performing waveform separation processing of the mass chromatogram of the internal standard substance, and 
 calculating an area of a waveform specific to the internal standard substance obtained from the waveform separation processing to compute a ratio (an area of a first waveform)/(an area of a waveform specific to the internal standard substance), and/or, a ratio (an area of a second waveform)/(an area of a waveform specific to the internal standard substance). 
 
     
     
         74 . A program for causing a computer receiving data obtained in a mass spectroscopic device to analyze the data in which the data is related to mass analysis of a first and a second biological molecule, and an internal standard substance, which is a biological molecule that has been isotope labeled for corresponding to the first and the second biological molecules, the program performing the steps of:
 determining, among the data on the mass analysis of the biological molecules measured in the mass spectroscopic device, a first m/z value and a second m/z value of the first and the second biological molecules for which a sequence of a biological molecular component has been determined, and a m/z value of the biological molecule constituting the internal standard substance for which a sequence of a biological molecular component has been determined;   extracting data containing a mass spectrum against m/z values and a mass chromatogram of the first and the second biological molecules as well as of the internal standard substance biological molecule from the data on the mass analysis, based on the m/z values of the first and the second as well as of the internal standard substance; and   performing waveform separation processing of the mass chromatogram of the first and second of the biological molecules, as well as the biological molecule constituting the internal standard substance.   
     
     
         75 . The program according to  claim 74 , wherein the determination of the first m/z value and the second m/z value, as well as the m/z value of the internal standard substance is carried out in such was that the determination of the m/z value of the biological molecule constituting the internal standard substance is made by determining, from the first m/z value and the second m/z value of the first and the second biological molecules for which a sequence of the biological molecular component has been determined, based on information on the number of non-isotope labeled components of the first and the second biological molecules and an electric charge of the first and the second biological molecules, or the determination of the first m/z value and the second m/z value is made by determining, from the m/z value of the internal standard substance for which a sequence of the biological molecular component has been determined, based on information on the number of isotope labeled biological molecular components of the internal standard substance and an electric charge of the biological molecule constituting the internal standard substance. 
     
     
         76 . The program according to  claim 74  or  75 , further comprising performing the steps of:
 calculating an area of a waveform derived from an individual biological molecule obtained by the waveform separation processing step; and 
 performing calculation a ratio (an area of a first waveform)/(an area of a waveform specific to the internal standard substance), and/or, a ratio (an area of a second waveform)/(an area of a waveform specific to the internal standard substance) from values of the areas of the first and the second waveforms derived from specific biological molecules at the first and the second m/z values, and a value of the area of the internal standard substance waveform derived from an individual biological molecule at m/z value of the internal standard substance. 
 
     
     
         77 . The program according to any one of  claims 74  to  76 , further comprising performing the steps of:
 performing, after the waveform separation, classification for each sequence of the component of the biological molecule; 
 retrieving the sequence with a database containing information on the function of biological molecules; and 
 linking the sequence to function from the result of the verification so as to group. 
 
     
     
         78 . The program according to any one of  claims 74  to  77 , further comprising performing the steps of:
 identifying the biological molecule from the sequence of the biological molecular component; and 
 determining, from a quantitative value of each biological molecular component constituting the biological molecule, a mean value to serve as a representative quantitative value for the biological molecule. 
 
     
     
         79 . The program according to any one of  claims 74  to  78 , further comprising performing of retrieving the sequence of the biological molecular component with a database containing information on the function of biological molecules so as to acquire information on the function of the biological molecule. 
     
     
         80 . The program according to any one of  claims 67  to  79 , wherein the isotope used in the isotope labeling is any isotope selected from the group consisting of  2 H,  13 C,  15 N,  17 O,  18 O,  33 P and  34 S, as well as combinations thereof. 
     
     
         81 . The program according to any one of  claims 67  to  80 , wherein the determination of the sequence of the component is carried out by MS/MS processing. 
     
     
         82 . The program according to  claim 81 , wherein the determination of the sequence of the component is carried out by retrieving data obtained by MS/MS processing with a database regarding the biological molecule. 
     
     
         83 . The program according to any one of  claims 67  to  82 , wherein the biological molecule is a biological molecule that has been separated from the sample over time. 
     
     
         84 . The program according to any one of  claims 67  to  83 , wherein the biological molecule is any molecule selected from the group consisting of a protein, a lipid, a sugar chain and a nucleic acid, as well as combinations thereof. 
     
     
         85 . An analysis method for analyzing data on mass analysis of an internal standard substance, which is a biological molecule that has been isotope labeled for corresponding to a first and a second biological molecule, comprising the steps of:
 determining, among data on the mass analysis of the biological molecule measured in a mass spectroscopic device, a first m/z value and a second m/z value of the first and the second biological molecules for which a sequence of a biological molecular component has been determined, and a m/z value of a biological molecule constituting an internal standard substance for which a sequence of a biological molecular component has been determined; and   retrieving with a database containing information on the function of biological molecules based on the obtained data, so as to acquire information on the function of the biological molecule.   
     
     
         86 . The analysis method, further comprising, after the determination step, extracting a mass spectrum against m/z values and a mass chromatogram of the first and the second biological molecules as well as of the biological molecule constituting the internal standard substance, from the data on the mass analysis, based on the first and the second m/z values as well as the m/z value of the internal standard substance. 
     
     
         87 . The analysis method according to  claim 85  or  86 , wherein the determination of the first m/z value and the second m/z value, as well as the m/z value of the internal standard substance is carried out in such was that the determination of the m/z value of the biological molecule constituting the internal standard substance is made by determining, from the first m/z value and the second m/z value of the first and the second biological molecules for which a sequence of the biological molecular component has been determined, based on information on the number of non-isotope labeled components of the first and the second biological molecules and an electric charge of the first and the second biological molecules, or the determination of the first m/z value and the second m/z value is made by determining, from the m/z value of the internal standard substance for which a sequence of the biological molecular component has been determined, based on information on the number of isotope labeled biological molecular components of the internal standard substance and an electric charge of the biological molecule constituting the internal standard substance. 
     
     
         88 . The analysis method according to any one of  claims 85  to  87 , further comprising performing calculation of a ratio (a first peak)/(a peak specific to the internal standard substance), and/or, a ratio (a second peak)/(a peak specific to the internal standard substance) from values of the first and the second peaks derived from specific biological molecules at the first and the second m/z values, and a value of an internal standard substance peak derived from an individual biological molecule at the m/z value of the internal standard substance. 
     
     
         89 . The analysis method according to any one of  claims 85  to  88 , further comprising linking the sequence to function from the result of the verification so as to group. 
     
     
         90 . The analysis method according to any one of  claims 85  to  89 , further comprising the steps of:
 identifying the biological molecule from the sequence of the biological molecular component; and 
 determining, from a quantitative value of each biological molecular component constituting the biological molecule, a mean value to serve as a representative quantitative value for the biological molecule. 
 
     
     
         91 . The analysis method according to any one of  claims 85  to  90 , wherein the isotope used in the isotope labeling is any isotope selected from the group consisting of  2 H,  13 C,  15 N,  17 O,  18 O,  33 P and  34 S, as well as combinations thereof 
     
     
         92 . The analysis method according to any one of  claims 85  to  91 , wherein the determination of the sequence of the component is carried out by MS/MS processing. 
     
     
         93 . The analysis method according to  claim 92 , wherein the determination of the sequence of the component is carried out by retrieving data obtained by MS/MS processing with a database regarding a biological molecule. 
     
     
         94 . The analysis method according to any one of  claims 85  to  93 , wherein the biological molecule is a biological molecule that has been separated from the sample over time. 
     
     
         95 . The analysis method according to any one of  claims 85  to  94 , wherein the biological molecule is any molecule selected from the group consisting of a protein, a lipid, a sugar chain, and a nucleic acid, as well as combinations thereof. 
     
     
         96 . A program for causing a computer receiving data obtained in a mass spectroscopic device to analyze the data in which the data is related to mass analysis of a first and a second biological molecule, and an internal standard substance, which is a biological molecule that has been isotope labeled for corresponding to the first and the second biological molecules, the program performing the steps of:
 determining, among the data on the mass analysis of the biological molecule measured in the mass spectroscopic device, a first m/z value and second m/z value of the first and the second biological molecules for which a sequence of a biological molecular component has been determined, and a m/z value of a biological molecule constituting an internal standard substance for which a sequence of a biological molecular component has been determined,   retrieving with a database containing information on the function of biological molecules based on the obtained data, so as to acquire information on the function of the biological molecule.   
     
     
         97 . The program according to  claim 96 , further comprising performing of, after the determination step, extracting a mass spectrum against m/z values and a mass chromatogram of the first and the second biological molecules as well as of the biological molecule constituting the internal standard substance, from the data on the mass analysis, based on the first and the second m/z values as well as the m/z value of the internal standard substance. 
     
     
         98 . The program according to  claim 96  or  97 , wherein the determination of the first m/z value and the second m/z value, as well as the m/z value of the internal standard substance is carried out in such was that the determination of the m/z value of the biological molecule constituting the internal standard substance is made by determining, from the first m/z value and the second m/z value of the first and the second biological molecules for which a sequence of the biological molecular component has been determined, based on information on the number of non-isotope labeled components of the first and the second biological molecules and an electric charge of the first and the second biological molecules, or the determination of the first m/z value and the second m/z value is made by determining, from the m/z value of the internal standard substance for which a sequence of the biological molecular component has been determined, based on information on the number of isotope labeled biological molecular components of the internal standard substance and an electric charge of the biological molecule constituting the internal standard substance. 
     
     
         99 . The program according to any one of  claims 96  to  98 , further comprising performing calculation of a ratio (a first peak)/(a peak specific to the internal standard substance), and/or, a ratio (a second peak)/(a peak specific to the internal standard substance) from values of the first and the second peaks derived from specific biological molecules at the first and the second m/z values, and a value of an internal standard substance peak derived from an individual biological molecule at the m/z value of the internal standard substance. 
     
     
         100 . The program according to any one of  claims 96  to  99 , further comprising performing of linking the sequence to function from the result of the verification so as to group. 
     
     
         101 . The program according to any one of  claims 96  to  100 , further comprising performing the steps of:
 identifying the biological molecule from the sequence of the biological molecular component; and 
 determining, from a quantitative value of each biological molecular component constituting the biological molecule, a mean value to serve as a representative quantitative value for the biological molecule. 
 
     
     
         102 . The program according to any one of  claims 96  to  101 , wherein the isotope used in the isotope labeling is any isotope selected from the group comprising  2 H, C,  15 N,  17 O,  18 O,  33 P and  34 S, as well as combinations thereof. 
     
     
         103 . The program according to any one of  claims 96  to  102 , wherein the determination of the sequence of the component is carried out by MS/MS processing. 
     
     
         104 . The program according to  claim 103 , wherein the determination of the sequence of the component is carried out by retrieving data obtained by MS/MS processing with a database regarding a biological molecule. 
     
     
         105 . The program according to any one of  claims 96  to  104 , wherein the biological molecule is a biological molecule that has been separated from the sample over time. 
     
     
         106 . The program according to any one of  claims 96  to  105 , wherein the biological molecule is any molecule selected from the group consisting of a protein, a lipid, a sugar chain and a nucleic acid, as well as combinations thereof. 
     
     
         107 . A program for causing a computer receiving data on mass analysis obtained in a mass spectroscopic device in which the date is related to one or a plurality of biological molecules in a sample, to quantitate the one or a plurality of biological molecules in the sample, the program performing the steps of
 determining, among the data on the mass analysis of the biological molecule measured in the mass spectroscopic device, a first m/z value of the first biological molecule for which a sequence of a biological molecular component has been determined, and a second m/z value of the second biological molecule for which a sequence of a biological molecular component has been determined,   extracting data containing a mass spectrum against m/z values and a mass chromatogram of the first biological molecule and the second biological molecule from the data on the mass analysis, based on the first and the second m/z values; and   performing waveform separation processing of the mass chromatogram of the first and the second biological molecules.   
     
     
         108 . The program according to  claim 107 , wherein the determination of the first m/z value and the second m/z value is carried out in such a way that the determination of the second m/z value is made by determining from the first m/z value of the first biological molecule for which the sequence of the biological molecular component has been determined, based on information on the number of non-isotope labeled components of the first biological molecule and an electric charge of the first biological molecule, or the determination of the first m/z value of the second biological molecule is made by determining from the second m/z value of the second biological molecule for which the sequence of the biological molecular component has been determined, based on information on the number of isotope labeled components of the second biological molecule and an electric charge of the second biological molecule. 
     
     
         109 . The program according to  claim 107  or  108 , further comprising performing the steps of:
 calculating an area of a waveform derived from an individual biological molecule obtained by the waveform separation processing step; and 
 calculating a ratio (an area of a first waveform)/(an area of a second waveform) from a value of the area of the first waveform derived from an individual biological molecule at the first m/z value, and a value of the area of the second waveform derived from an individual biological molecule at the second m/z value. 
 
     
     
         110 . The program according to any one of  claims 107  to  109 , further comprising performing the steps of:
 performing, after the waveform separation, classification for each sequence of a component of the biological molecule, 
 retrieving the sequence with a database containing information on the function of biological molecules, and 
 linking the sequence to function from the result of the verification so as to group. 
 
     
     
         111 . The program according to any one of  claims 107  to  110 , further comprising performing the steps of:
 identifying the biological molecule from the sequence of the biological molecular component; and 
 determining, from a quantitative value of each biological molecular component constituting the biological molecule, a mean value to serve as a representative quantitative value for the biological molecule. 
 
     
     
         112 . The program according to any one of  claims 107  to  111 , further comprising performing of retrieving the sequence of the biological molecular component with a database containing information on the function of biological molecules so as to acquire information on the function of the biological molecule. 
     
     
         113 . The program according to any one of  claims 107  to  112 , wherein the isotope used in the isotope labeling is any isotope selected from the group comprising  2 H,  13 C,  15 N,  17 O,  18 O,  33 P and  34 S, as well as combinations thereof. 
     
     
         114 . The program according to any one of  claims 107  to  113 , wherein the determination of the sequence of the component is carried out by MS/MS processing. 
     
     
         115 . The program according to  claim 114 , wherein the determination of the sequence of the component is carried out by retrieving data obtained by MS/MS processing with a database regarding a biological molecule. 
     
     
         116 . The program according to any one of  claims 107  to  115 , wherein the biological molecule is a biological molecule that has been separated from the sample over time. 
     
     
         117 . The program according to any one of  claims 107  to  116 , wherein the biological molecule is any molecule selected from the group consisting of a protein, a lipid, a sugar chain and a nucleic acid, as well as combinations thereof.