CN115932142A - Spectrogram analysis method and device - Google Patents

Abstract

The invention discloses a spectrogram analysis method and a spectrogram analysis device, wherein the spectrogram analysis method comprises the following standard component analysis processes: positioning the peak position of each chromatographic peak in the sample chromatogram, and acquiring a mixed mass spectrogram of the peak positions; filtering invalid mass spectrum peaks in the mixed mass spectrum to obtain a first mass spectrum, and matching the first mass spectrum in a database to obtain a plurality of matching results with scores ranked first; calculating the final score of each standard component, recording the standard component corresponding to the highest final score as a first component, and performing component deduction processing on the first component in the first mass spectrogram and the chromatographic peak thereof; recalculating chromatographic peaks of the first mass spectrum and repeating the standard component analysis process until the sample chromatogram no longer contains components. The invention improves the anti-noise interference capability of the chromatographic peak analysis technology and widens the application range of the chromatographic peak analysis technology.

Description

Spectrogram analysis method and device

Technical Field

The invention relates to the technical field of chromatographic peak analysis, in particular to a spectrogram analysis method and device.

Background

The chromatogram has strong separating capacity and poor qualitative capacity to unknown compounds; the mass spectrum has unique identification capability on unknown compounds and extremely high sensitivity, but the detected components are generally required to be pure compounds. The combination of the chromatogram and the mass spectrum, namely the combination of the chromaticness and the mass spectrum, makes good use of the advantages and the disadvantages of each other, makes up for the defect that the chromatogram is difficult to perform reliable qualitative identification on unknown components in complex compounds, and utilizes the mass spectrum with strong identification capability and extremely high sensitivity as a detector. The existing chromatography-mass spectrometry technology has certain defects and limited separation capacity for components with similar structures and close co-efflux peak time. The comprehensive two-dimensional chromatographic analysis technology relies on two chromatographic columns which have different separation mechanisms and are independent of each other (the two chromatographic columns are connected in series), and components which are not separated in the first dimension are further separated in the second dimension, so that the chromatographic separation capacity is improved. However, in a complex mixture, a large number of components inevitably cause mass spectrum overlapping, which is reflected in the chromatogram: a plurality of components are mixed together, and the number of chromatographic peaks on the chromatogram is smaller than that of the components, even only one chromatographic peak is provided.

In the prior art, some separation methods have been proposed to further improve the chromatographic separation and the resolution of the co-effluent peak. For example, the method for analyzing the co-effluent peak and searching the spectral library based on the gas chromatography-mass spectrometry proposed by the Luhong soldier and the like utilizes a three-dimensional omnibearing spectrogram and a method for comparing the combination of mass spectrum data corresponding to a chromatographic peak and spectral library sample data, increases the dimension of separation by means of three-dimensional information, and improves the separation capability. However, in this method, the component number needs to be determined according to the peak shape and the peak purity, it is difficult to determine a chromatographic peak with a high co-flow peak overlapping degree, the interference resistance is weak, and the mass spectrum to be analyzed needs to be combined and compared with the mass spectrum in the spectrum library, which results in a very large amount of calculation. The prior art also includes methods such as PCA and PALS, which generally need to satisfy some conditions, such as a certain function that the peak shape must satisfy, and the application range is narrow.

Disclosure of Invention

The invention provides a spectrogram analysis method, which solves the problems of weak noise interference resistance and narrow application range in the existing chromatographic peak analysis technology, improves the noise interference resistance of the chromatographic peak analysis technology by separating a chromatographic spectrum step by step, recalculating the chromatographic peak after subtracting corresponding components once per separation and separating the next-stage chromatogram, and widens the application range.

In order to solve the above technical problem, an embodiment of the present invention provides a method for analyzing a spectrogram, including the following standard component analysis processes:

positioning the peak position of each chromatographic peak in the sample chromatogram, and acquiring a mixed mass spectrogram of the peak positions;

filtering invalid mass spectrum peaks in the mixed mass spectrum to obtain a first mass spectrum, and matching the first mass spectrum in a database to obtain a plurality of matching results with scores ranked first, wherein the matching results comprise standard component names, standard mass spectra and matching item scores;

calculating the final score of each standard component, recording the standard component corresponding to the highest final score as a first component, and performing component deduction processing on the first component in the first mass spectrogram and the chromatographic peak thereof;

recalculating chromatographic peaks of the first mass spectrum and repeating the standard component analysis process until the sample chromatogram no longer contains components.

As one preferable scheme, the step of subtracting the first component from the first mass spectrum and the chromatographic peak thereof specifically comprises:

and taking the maximum peak ion in the standard mass spectrogram of the first component as a maximum scale factor, calculating the scale factor and ion deduction intensity of the peak ion in the standard mass spectrogram from large to small in sequence according to the intensity of the peak ion, and deducting corresponding ions from the first mass spectrogram in sequence according to the ion deduction intensity.

As one preferable scheme, the calculating the final score of each standard component specifically includes:

and calculating the final score according to the number of the standard components in the matching result, the average score of the current standard components and the occurrence times of the current standard components.

As one of the preferable schemes, the final score is calculated according to the following formula:

wherein f represents the final score, n is the number of occurrences of the current standard component,

k represents the number of standard components in the matching result, which is the average score of the current standard components.

As one of the preferred schemes, the matching item score includes a forward matching score, a reverse matching score and a probability score.

As one of the preferable schemes, the matching results which do not satisfy the effective condition are eliminated before the final scores of the respective standard components are calculated.

As one of the preferable schemes, the removing the matching result which does not satisfy the valid condition specifically includes:

when the sum of the forward matching score and the reverse matching score is greater than 1000 or the probability is greater than 50%, the validity condition is satisfied.

The invention provides a spectrogram analyzing device, which comprises a mixed mass spectrogram acquiring module, an invalid mass spectrogram peak filtering module, a component deduction module and a circulation module, wherein the mixed mass spectrogram acquiring module is used for acquiring a mass spectrogram;

the module is used for positioning the peak position of each chromatographic peak in the sample chromatogram and acquiring a mixed mass spectrogram of the peak position;

the invalid mass spectrum peak filtering module is used for filtering invalid mass spectrum peaks in the mixed mass spectrum to obtain a first mass spectrum, matching the first mass spectrum in a database to obtain a plurality of matching results with scores ranked first, wherein the matching results comprise standard component names, standard mass spectrum and matching item scores;

the component deduction module is used for calculating the final score of each standard component, marking the standard component corresponding to the highest final score as a first component, and carrying out component deduction processing on the first component in the first mass spectrogram and a chromatographic peak thereof;

the cycle module is configured to recalculate the chromatographic peak of the first mass spectrum and to repeatedly perform the standard component analysis procedure until the sample chromatogram no longer contains a component.

Compared with the prior art, the embodiment of the invention has the advantages that at least one point is as follows:

the invention can accurately determine the number of the co-outflow peaks and the specific component information by separating the chromatogram step by step, deducting the corresponding components once per separation, recalculating the chromatographic peak, and separating the next-stage chromatogram. Meanwhile, the method can realize the effect of effectively providing the co-outflow peak analysis capability of the chromatography-mass spectrometry combined analysis without upgrading hardware facilities. The chromatographic peak analysis technology of the invention has strong anti-noise interference capability and is suitable for chromatographic data generated by most chromatographic instruments.

Drawings

FIG. 1 is a schematic flow chart of a chromatographic peak analysis method with step-by-step separation in one embodiment of the present invention;

FIG. 2 is a schematic diagram of a chromatographic peak analyzer with step-by-step separation in one embodiment of the present invention;

FIG. 3 is a schematic diagram of the chromatographic peak profile of the co-eluting peak of the chromatographic peak resolution method for separation by stage in one embodiment of the present invention;

FIG. 4 is a schematic three-dimensional mass spectrum of the chromatogram peak profile of the co-elution peak of the chromatographic peak resolution method for stepwise separation in one embodiment of the present invention;

fig. 5 is a schematic diagram of a mixed mass spectrum corresponding to a chromatographic peak position of a chromatographic peak resolution method for step-by-step separation in an embodiment of the present invention.

Detailed Description

The technical solutions in the embodiments of the present invention will be described clearly and completely with reference to the accompanying drawings in the embodiments of the present invention, and it is to be understood that the described embodiments are only a part of the embodiments of the present invention, and not all of the embodiments, and the embodiments are provided for the purpose of making the disclosure of the present invention more thorough and complete. All other embodiments, which can be derived by a person skilled in the art from the embodiments given herein without making any creative effort, shall fall within the protection scope of the present invention.

In the description of the present application, the terms "first", "second", "third", etc. are used for descriptive purposes only and are not to be construed as indicating or implying relative importance or implying any number of technical features indicated. Thus, features defined as "first," "second," "third," etc. may explicitly or implicitly include one or more of the features. In the description of the present application, "a plurality" means two or more unless otherwise specified.

In the description of the present application, it is to be noted that, unless otherwise explicitly specified or limited, the terms "mounted," "connected," and "connected" are to be construed broadly, e.g., as meaning either a fixed connection, a removable connection, or an integral connection; can be mechanically or electrically connected; they may be connected directly or indirectly through intervening media, or they may be interconnected between two elements. As used herein, the terms "vertical," "horizontal," "left," "right," "up," "down," and the like are for illustrative purposes only and do not indicate or imply that the referenced device or element must be in a particular orientation, constructed or operated in a particular manner, and is not to be construed as limiting the present invention. As used herein, the term "and/or" includes any and all combinations of one or more of the associated listed items. The specific meaning of the above terms in this application will be understood to be a specific case for those of ordinary skill in the art.

In the description of the present application, it is to be noted that, unless defined otherwise, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this invention belongs. The terminology used in the description of the invention herein is for the purpose of describing particular embodiments only and is not intended to be limiting of the invention, as those skilled in the art will recognize the specific meaning of the terms used in the present application in a particular context.

An embodiment of the present invention provides a method for analyzing chromatographic peaks separated step by step, please refer to fig. 1, where fig. 1 is a schematic flow chart of the method for analyzing chromatographic peaks separated step by step in an embodiment of the present invention, and the method includes:

step S1: and positioning the peak position of each chromatographic peak in the sample chromatogram, and acquiring a mixed mass spectrogram of the peak positions. Specifically, referring to fig. 3 and 4, fig. 3 shows the chromatogram peak patterns of the co-elution peak before the chromatographic separation, and the chromatogram peak patterns of the respective components analyzed by the present method. Judging whether each chromatographic peak in a sample chromatogram is a co-outflow peak or not according to the chromatographic peak pattern (figure 3) of the co-outflow peak and a corresponding three-dimensional mass spectrogram (figure 4), if so, locating the peak position in the co-outflow peak in the sample chromatogram, wherein the peak position is preferably 2.35 min-2.4 min; referring to fig. 5, a mixed mass spectrum corresponding to the peak position is obtained (fig. 5).

Step S2: and filtering invalid mass spectrum peaks in the mixed mass spectrum to obtain a first mass spectrum, and matching the first mass spectrum in a database to obtain a plurality of matching results with scores ranked first, wherein the matching results comprise standard component names, standard mass spectra and matching item scores. The database is a standard spectrum in a mass spectrometer.

As one example, the invalid mass spectrum peak in the mixed mass spectrum is filtered to obtain a first mass spectrum; specifically, peak ions with the intensity smaller than a first threshold value in the mixed mass spectrogram are removed, and interference is avoided. Preferably, the first threshold =0.01 × peak ion maximum intensity.

As an embodiment, the first mass spectrogram is matched in a nist database to obtain a matching result with a score of 5, the matching result comprises a standard component name, a standard mass spectrogram and a matching item score, and the matching item comprises a forward matching score, a reverse matching score and a probability score.

And step S3: calculating the final score of each standard component, recording the standard component corresponding to the final score with the first ranking as a first component, and carrying out component deduction processing on the first component in the first mass spectrogram and the chromatographic peak thereof.

As one of the embodiments, the matching results that do not satisfy the valid condition are rejected before the final scores of the respective standard components are calculated. The method specifically comprises the following steps: when the sum of the forward matching score and the reverse matching score is greater than 1000 or the probability is greater than 50%, the validity condition is satisfied. If not, the valid condition is not satisfied, and the result is considered invalid. Two invalid results are excluded from the top 5 ranked matching results, leaving 3 matching results as follows:

molecular formula	Forward matching	Reverse matching	Probability of
				CH4O	916	924	98.6
CH4O	908	915	97
				C2H4O3	886	886	89.2

According to the formula

The final score of the standard ingredient CH40 is->

The final score of the standard component C2H4O3 is ^ based>

And recording a standard component C2H4O3 corresponding to the final score with the first ranking as a first component, and carrying out component deduction processing on the first component in the first mass spectrogram and a chromatographic peak thereof.

Wherein f represents the final score, n is the number of occurrences of the current standard component,

k represents the number of standard components in the matching result, which is the average score of the current standard components.

As an embodiment, the maximum peak ion in the standard mass spectrum of the first component is used as the maximum scale factor, the scale factor and the ion subtraction intensity of the peak ion in the standard mass spectrum are sequentially calculated from large to small according to the intensity of the peak ion, and then the corresponding ion is sequentially subtracted from the first mass spectrum according to the ion subtraction intensity. For example, the maximum peak ion (number 49) in the standard mass spectrum is used as the maximum scale factor 1, the ion subtraction intensity is 1, and the corresponding peak ion is subtracted from the first mass spectrum according to the ion subtraction intensity 1; calculating the next subtracted peak ion (numbered 84, intensity row two), with a scaling factor of 61.2/100=0.612, with an ion subtraction intensity of m x 0.612, m being, and subtracting the corresponding peak ion from the first mass spectrum according to the ion subtraction intensity of 0.612; the next subtracted peak ion (No. 51, intensity third), with a scaling factor of 35.9/100=0..359, with an ion subtracted intensity of m × 0.359, the corresponding peak ion was subtracted from the first mass spectrum according to the ion subtracted intensity of 0.359; and so on until the last peak ion is deducted.

And step S4: recalculating chromatographic peaks of the first mass spectrum and repeating the standard component analysis process until the sample chromatogram no longer contains components.

The invention can accurately determine the number of the co-outflow peaks and the specific component information by separating the chromatogram step by step, deducting the corresponding components once per separation, recalculating the chromatographic peaks and then separating the next-stage chromatogram. Meanwhile, the method can realize the effect of effectively providing the co-outflow peak analysis capability of the chromatography-mass spectrometry combined analysis without upgrading hardware facilities. The method filters the invalid mass spectrum peak in the mixed mass spectrum to obtain the first mass spectrum, so that the method has strong anti-noise interference capability and is suitable for chromatographic data generated by most chromatograph-mass spectrometer instruments.

Another embodiment of the present invention is a chromatographic peak analysis device with step-by-step separation, please refer to fig. 2, fig. 2 is a schematic structural diagram of the chromatographic peak analysis device with step-by-step separation in one embodiment of the present invention, which includes a mixed mass spectrum acquisition module 101, an invalid mass spectrum peak filtering module 102, a component deduction module 103, and a next component analysis module 104;

the mixed mass spectrogram acquisition module is used for positioning the peak position of each chromatographic peak in the sample chromatogram and acquiring the mixed mass spectrogram of the peak position;

the invalid mass spectrum peak filtering module is used for filtering invalid mass spectrum peaks in the mixed mass spectrum to obtain a first mass spectrum, matching the first mass spectrum in a database to obtain a plurality of matching results with scores ranked first, wherein the matching results comprise standard component names, standard mass spectrum and matching item scores;

the component deduction module is used for calculating the final score of each standard component, marking the standard component corresponding to the highest final score as a first component, and carrying out component deduction processing on the first component in the first mass spectrogram and a chromatographic peak thereof;

and the next composition analysis module is used for recalculating the chromatographic peak of the first mass spectrum, and repeatedly executing the standard component analysis process of the mixed mass spectrum acquisition module, the invalid mass spectrum peak filtering module and the component deduction module until the sample chromatogram does not contain any component.

It should be noted that the above device embodiment of the present invention corresponds to the method embodiment of the present invention, and the method for analyzing a chromatographic peak of a vehicle progressive separation according to any method embodiment of the present invention can be implemented.

The above-mentioned embodiments only express several embodiments of the present invention, and the description thereof is specific and detailed, but not to be understood as limiting the scope of the present invention. It should be noted that various changes and modifications can be made by those skilled in the art without departing from the spirit of the invention, and these changes and modifications are all within the scope of the invention. Therefore, the protection scope of the present patent shall be subject to the appended claims.

Claims (8)

1. The method for analyzing the spectrogram is characterized by comprising the following standard component analysis processes:

positioning the peak position of each chromatographic peak in the sample chromatogram, and acquiring a mixed mass spectrogram of the peak positions;

filtering invalid mass spectrum peaks in the mixed mass spectrum to obtain a first mass spectrum, and matching the first mass spectrum in a database to obtain a plurality of matching results with scores ranked first, wherein the matching results comprise standard component names, standard mass spectra and matching item scores;

calculating the final score of each standard component, recording the standard component corresponding to the highest final score as a first component, and performing component deduction processing on the first component in the first mass spectrogram and the chromatographic peak thereof;

recalculating chromatographic peaks of the first mass spectrum and repeating the standard component analysis process until the sample chromatogram no longer contains components.

2. The method for analyzing spectrum according to claim 1, wherein the step of subtracting the first component from the first mass spectrum and its peak comprises:

and taking the maximum peak ion in the standard mass spectrogram of the first component as a maximum scale factor, calculating the scale factor and ion deduction intensity of the peak ion in the standard mass spectrogram from large to small in sequence according to the intensity of the peak ion, and deducting corresponding ions from the first mass spectrogram in sequence according to the ion deduction intensity.

3. The method for spectrum analysis according to claim 2, wherein the calculating of the final score for each standard component comprises:

and calculating the final score according to the number of the standard components in the matching result, the average score of the current standard components and the occurrence times of the current standard components.

4. A method of parsing a spectrogram according to claim 3, wherein said final score is calculated according to the following formula:

wherein f represents the final score, n is the number of occurrences of the current standard component,

k represents the number of standard components in the matching result, which is the average score of the current standard components.

5. The method of parsing a spectrogram according to claim 4, wherein said match term scores comprise a forward match score, a reverse match score and a probability score.

6. A method of spectrogram according to claim 5, wherein said matching results which do not satisfy valid conditions are rejected before calculating the final score for each standard component.

7. The method for spectrogram parsing of any one of claims 1 to 6, wherein said rejecting said matching results that do not satisfy an effective condition specifically comprises:

when the sum of the forward matching score and the reverse matching score is greater than 1000 or the probability is greater than 50%, the validity condition is satisfied.

8. The device for analyzing the spectrogram is characterized by comprising a mixed mass spectrogram acquisition module, an invalid mass spectrogram peak filtering module, a component deduction module and a next component analysis module;

the mixed mass spectrum acquisition module is used for positioning the peak position of each chromatographic peak in the sample chromatogram and acquiring the mixed mass spectrum of the peak position;

the invalid mass spectrum peak filtering module is used for filtering invalid mass spectrum peaks in the mixed mass spectrum to obtain a first mass spectrum, matching the first mass spectrum in a database to obtain a plurality of matching results with scores ranked first, wherein the matching results comprise standard component names, standard mass spectrum and matching item scores;

the component deduction module is used for calculating the final score of each standard component, marking the standard component corresponding to the highest final score as a first component, and carrying out component deduction processing on the first component in the first mass spectrogram and a chromatographic peak thereof;

the next mass analysis module is configured to recalculate the chromatographic peak of the first mass spectrum and to repeat the standard component analysis process until the sample chromatogram contains no more components.

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