CN113466320B - MALDI-TOF detection method for bovine-derived components in milk powder - Google Patents
MALDI-TOF detection method for bovine-derived components in milk powder Download PDFInfo
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- CN113466320B CN113466320B CN202110903420.0A CN202110903420A CN113466320B CN 113466320 B CN113466320 B CN 113466320B CN 202110903420 A CN202110903420 A CN 202110903420A CN 113466320 B CN113466320 B CN 113466320B
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- 238000001514 detection method Methods 0.000 title claims abstract description 18
- 239000008267 milk Substances 0.000 title claims abstract description 15
- 210000004080 milk Anatomy 0.000 title claims abstract description 15
- 235000013336 milk Nutrition 0.000 title claims abstract description 15
- 239000000843 powder Substances 0.000 title claims abstract description 15
- 241000283690 Bos taurus Species 0.000 title claims abstract description 9
- 238000001840 matrix-assisted laser desorption--ionisation time-of-flight mass spectrometry Methods 0.000 title claims abstract description 9
- 239000011259 mixed solution Substances 0.000 claims abstract description 16
- 239000011159 matrix material Substances 0.000 claims abstract description 12
- 230000008859 change Effects 0.000 claims description 3
- 239000012634 fragment Substances 0.000 claims description 3
- 238000001819 mass spectrum Methods 0.000 claims description 3
- 238000000034 method Methods 0.000 claims description 2
- 230000007547 defect Effects 0.000 abstract 1
- 150000002500 ions Chemical class 0.000 description 54
- 238000012986 modification Methods 0.000 description 2
- 230000004048 modification Effects 0.000 description 2
- 230000009471 action Effects 0.000 description 1
- 230000009286 beneficial effect Effects 0.000 description 1
- 238000012937 correction Methods 0.000 description 1
- 238000003795 desorption Methods 0.000 description 1
- 238000010586 diagram Methods 0.000 description 1
- 230000005684 electric field Effects 0.000 description 1
- 238000000752 ionisation method Methods 0.000 description 1
- 230000001678 irradiating effect Effects 0.000 description 1
- 238000004949 mass spectrometry Methods 0.000 description 1
- 230000008569 process Effects 0.000 description 1
- 238000012545 processing Methods 0.000 description 1
- 238000001179 sorption measurement Methods 0.000 description 1
- 238000001228 spectrum Methods 0.000 description 1
- 239000000126 substance Substances 0.000 description 1
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- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N27/00—Investigating or analysing materials by the use of electric, electrochemical, or magnetic means
- G01N27/62—Investigating or analysing materials by the use of electric, electrochemical, or magnetic means by investigating the ionisation of gases, e.g. aerosols; by investigating electric discharges, e.g. emission of cathode
- G01N27/64—Investigating or analysing materials by the use of electric, electrochemical, or magnetic means by investigating the ionisation of gases, e.g. aerosols; by investigating electric discharges, e.g. emission of cathode using wave or particle radiation to ionise a gas, e.g. in an ionisation chamber
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- G06T5/70—
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- G06T5/80—
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- G—PHYSICS
- G06—COMPUTING; CALCULATING OR COUNTING
- G06F—ELECTRIC DIGITAL DATA PROCESSING
- G06F2218/00—Aspects of pattern recognition specially adapted for signal processing
- G06F2218/02—Preprocessing
- G06F2218/04—Denoising
-
- G—PHYSICS
- G06—COMPUTING; CALCULATING OR COUNTING
- G06F—ELECTRIC DIGITAL DATA PROCESSING
- G06F2218/00—Aspects of pattern recognition specially adapted for signal processing
- G06F2218/12—Classification; Matching
- G06F2218/14—Classification; Matching by matching peak patterns
Abstract
The invention discloses a MALDI-TOF detection method for bovine-derived components in milk powder, which comprises the following steps: A. respectively preparing a matrix mixed solution and a sample mixed solution by using a milk powder sample to be detected, and dropwise adding the matrix mixed solution and the sample mixed solution on a target plate; B. detecting the target plate by using MALDI-TOF equipment to obtain a mass-to-charge ratio peak spectrogram; C. and correcting each ion peak of the mass-to-charge ratio peak spectrogram. The invention can improve the defects of the prior art and improve the accuracy of MALDI-TOF detection on the milk powder.
Description
Technical Field
The invention relates to the technical field of chemical component detection, in particular to a MALDI-TOF (matrix-assisted laser desorption/ionization-time of flight) detection method for bovine-derived components in milk powder.
Background
Matrix assisted laser desorption ionization time of flight mass spectrometry (MALDI-TOF) is a cocrystallized film formed by irradiating a sample and a matrix with laser, the matrix absorbs energy from the laser and transfers the energy to biomolecules, the protons are transferred to the biomolecules or obtained from the biomolecules in the ionization process to ionize the biomolecules, the ions accelerate to fly through a flight tube under the action of an electric field, and the ions are detected according to different flight times of a detector, namely the mass-to-charge ratio of the ions is determined to be in direct proportion to the flight time of the ions, so that the ion detection is realized. In order to improve the accuracy of mass spectrometry detection of matrix-assisted laser adsorption ionization time-of-flight, the deviation caused by interference in the detection process needs to be corrected.
Disclosure of Invention
The invention aims to solve the technical problem of providing a MALDI-TOF detection method for bovine-derived components in milk powder, and improving the accuracy of MALDI-TOF detection on the milk powder.
In order to solve the technical problems, the technical scheme adopted by the invention is as follows.
A MALDI-TOF detection method for bovine-derived components in milk powder comprises the following steps:
A. respectively preparing a matrix mixed solution and a sample mixed solution by using a milk powder sample to be detected, and dropwise adding the matrix mixed solution and the sample mixed solution onto a target plate;
B. detecting the target plate by using MALDI-TOF equipment to obtain a mass-to-charge ratio peak spectrogram;
C. and correcting each ion peak in the mass-to-charge ratio peak spectrogram.
Preferably, the step C of correcting each ion peak of the mass-to-charge ratio peak spectrum comprises the steps of,
c1, determining the ion peak with the highest abundance as a base peak, and correcting the mass-to-charge ratio deviation of different sub-peaks in the base peak;
c2, determining the content of main elements in the sample to be detected according to the corrected base peak, and defining the preset range of the other elements according to the content of the main elements;
c3, correcting the ion peak to be corrected by using the corresponding standard ion peak for the ion peak falling into the preset range;
and C4, for the ion peak which does not fall into the preset range, determining a corresponding element through the mass-to-charge ratio range of the ion peak, and then correcting the ion peak to be corrected according to the corrected ion peak.
Preferably, in step C1, correcting for variations in mass-to-charge ratios of different sub-peaks in the base peak comprises the steps of,
c11, defining an effective range of a base peak according to a preset fullness change rate threshold;
c12, marking peak points and valley points in an effective range;
c13, calculating the distance between the adjacent peak points one by one;
and C14, setting an effective interval with the standard value as the center by taking the average value of the distances between the adjacent peak points as the standard value, reducing the distance between the adjacent peak points to the effective interval for the adjacent peak points outside the effective interval, and then adjusting the position of the valley point between the adjacent peak points to keep the distance deviation between the two peak points and the valley point smaller than the set threshold value.
Preferably, in step C3, the ion peak to be corrected is linearly adjusted so that the deviation ratio from the standard ion peak is minimized.
Preferably, the step C4 of correcting the ion peak to be corrected based on the corrected ion peak includes the steps of,
c41, judging the type of the ion peak to be corrected according to the corrected ion peak and the corresponding element of the ion peak to be corrected, wherein the type of the ion peak to be corrected comprises a fragment ion peak, a metastable ion peak and an isotope ion peak;
c42, performing secondary mass spectrum detection on the ion peak to be corrected, and adjusting the peak area of the obtained secondary characteristic peak to make the peak area of the obtained secondary characteristic peak be the same as that of the standard secondary characteristic peak;
and C43, denoising the adjusted secondary characteristic peak, and then recombining the secondary characteristic peak into a corrected ion peak.
Adopt the beneficial effect that above-mentioned technical scheme brought to lie in: according to the invention, on the premise of not increasing the equipment cost and complexity, different ion peaks are classified, and then targeted correction processing is carried out on the different ion peaks, so that interference factors in mass-to-charge ratio peak spectrograms are effectively removed.
Drawings
FIG. 1 is a schematic diagram of one embodiment of the present invention.
Detailed Description
One embodiment of the present invention comprises the steps of:
A. respectively preparing a matrix mixed solution and a sample mixed solution by using a milk powder sample to be detected, and dropwise adding the matrix mixed solution and the sample mixed solution onto a target plate;
B. detecting the target plate by using MALDI-TOF equipment to obtain a mass-to-charge ratio peak spectrogram;
C. and correcting each ion peak of the mass-to-charge ratio peak spectrogram.
In the step C, the step of correcting each ion peak of the mass-to-charge ratio peak spectrogram comprises the following steps,
c1, determining the ion peak with the highest abundance as a base peak, and correcting the mass-to-charge ratio deviation of different sub-peaks in the base peak;
c2, determining the content of main elements in the sample to be detected according to the corrected base peak, and defining the preset range of the rest elements according to the content of the main elements;
c3, correcting the ion peak to be corrected by using the corresponding standard ion peak for the ion peak falling into the preset range;
and C4, for the ion peak which does not fall into the preset range, determining a corresponding element through the mass-to-charge ratio range of the ion peak, and then correcting the ion peak to be corrected according to the corrected ion peak.
In step C1, correcting the mass-to-charge ratio deviations of different sub-peaks in the base peak comprises the steps of,
c11, defining an effective range of a base peak according to a preset fullness change rate threshold;
c12, marking peak points and valley points in an effective range;
c13, calculating the distance between the adjacent peak points one by one;
and C14, setting an effective interval with the standard value as the center by taking the average value of the distances between the adjacent peak points as the standard value, reducing the distance between the adjacent peak points to the effective interval for the adjacent peak points outside the effective interval, and then adjusting the positions of the valley points between the adjacent peak points to keep the distance deviation between the two peak points and the valley points smaller than a set threshold value.
And C3, linearly adjusting the ion peak to be corrected to minimize the deviation rate of the ion peak to be corrected and the standard ion peak.
In step C4, the step of correcting the ion peak to be corrected according to the corrected ion peak comprises the following steps,
c41, judging the type of the ion peak to be corrected according to the corrected ion peak and the corresponding element of the ion peak to be corrected, wherein the type of the ion peak to be corrected comprises a fragment ion peak, a metastable ion peak and an isotope ion peak;
c42, performing secondary mass spectrum detection on the ion peak to be corrected, and adjusting the peak area of the obtained secondary characteristic peak to make the peak area of the obtained secondary characteristic peak be the same as that of the standard secondary characteristic peak;
and C43, denoising the adjusted secondary characteristic peak, and then recombining the secondary characteristic peak into a corrected ion peak.
In the description of the present invention, it is to be understood that the terms "longitudinal", "lateral", "upper", "lower", "front", "rear", "left", "right", "vertical", "horizontal", "top", "bottom", "inner", "outer", and the like, indicate orientations or positional relationships based on those shown in the drawings, are merely for convenience of description of the present invention, and do not indicate or imply that the referenced devices or elements must have a particular orientation, be constructed and operated in a particular orientation, and thus, are not to be construed as limiting the present invention.
The foregoing shows and describes the general principles and broad features of the present invention and advantages thereof. It will be understood by those skilled in the art that the present invention is not limited to the embodiments described above, which are given by way of illustration of the principles of the present invention, but that various changes and modifications may be made without departing from the spirit and scope of the invention, and such changes and modifications are within the scope of the invention as claimed. The scope of the invention is defined by the appended claims and equivalents thereof.
Claims (3)
1. A MALDI-TOF detection method for bovine-derived components in milk powder is characterized by comprising the following steps:
A. respectively preparing a matrix mixed solution and a sample mixed solution by using a milk powder sample to be detected, and dropwise adding the matrix mixed solution and the sample mixed solution onto a target plate;
B. detecting the target plate by using MALDI-TOF equipment to obtain a mass-to-charge ratio peak spectrogram;
C. the method for correcting each ion peak of a mass-to-charge ratio peak spectrogram comprises the following steps,
c1, determining the ion peak with the highest abundance as a base peak, and correcting the mass-to-charge ratio deviation of different sub-peaks in the base peak comprises the following steps,
c11, defining an effective range of a base peak according to a preset fullness change rate threshold;
c12, marking peak points and valley points in an effective range;
c13, calculating the distance between the adjacent peak points one by one;
c14, setting an effective interval with the standard value as the center by taking the average value of the distances between the adjacent peak points as the standard value, and for the adjacent peak points outside the effective interval, firstly reducing the distance between the adjacent peak points to the effective interval, and then carrying out position adjustment on the valley point between the adjacent peak points, and keeping the distance deviation between the two peak points and the valley point smaller than the set threshold value;
c2, determining the content of main elements in the sample to be detected according to the corrected base peak, and defining the preset range of the rest elements according to the content of the main elements;
c3, correcting the ion peak to be corrected by using the corresponding standard ion peak for the ion peak falling into the preset range;
and C4, for the ion peak which does not fall into the preset range, determining a corresponding element through the mass-to-charge ratio range of the ion peak, and then correcting the ion peak to be corrected according to the corrected ion peak.
2. The MALDI-TOF detection method for bovine-derived components in milk powder according to claim 1, characterized in that: and C3, linearly adjusting the ion peak to be corrected to minimize the deviation rate of the ion peak to be corrected and the standard ion peak.
3. The MALDI-TOF detection method for bovine-derived component in milk powder according to claim 2, characterized in that: in the step C4, the step of correcting the ion peak to be corrected according to the corrected ion peak comprises the following steps,
c41, judging the type of the ion peak to be corrected according to the corrected ion peak and the corresponding element of the ion peak to be corrected, wherein the type of the ion peak to be corrected comprises a fragment ion peak, a metastable ion peak and an isotope ion peak;
c42, performing secondary mass spectrum detection on the ion peak to be corrected, and adjusting the peak area of the obtained secondary characteristic peak to make the peak area of the obtained secondary characteristic peak be the same as that of the standard secondary characteristic peak;
and C43, denoising the adjusted secondary characteristic peak, and then recombining the secondary characteristic peak into a corrected ion peak.
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