CN105929052A - Establishing method for high-resolution mass spectrum database of anti-oxidants and antiseptic, and detection method for anti-oxidants and antiseptic - Google Patents
Establishing method for high-resolution mass spectrum database of anti-oxidants and antiseptic, and detection method for anti-oxidants and antiseptic Download PDFInfo
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- CN105929052A CN105929052A CN201610248071.2A CN201610248071A CN105929052A CN 105929052 A CN105929052 A CN 105929052A CN 201610248071 A CN201610248071 A CN 201610248071A CN 105929052 A CN105929052 A CN 105929052A
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- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N30/00—Investigating or analysing materials by separation into components using adsorption, absorption or similar phenomena or using ion-exchange, e.g. chromatography or field flow fractionation
- G01N30/02—Column chromatography
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- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N30/00—Investigating or analysing materials by separation into components using adsorption, absorption or similar phenomena or using ion-exchange, e.g. chromatography or field flow fractionation
- G01N30/02—Column chromatography
- G01N30/04—Preparation or injection of sample to be analysed
- G01N30/06—Preparation
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- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N30/00—Investigating or analysing materials by separation into components using adsorption, absorption or similar phenomena or using ion-exchange, e.g. chromatography or field flow fractionation
- G01N30/02—Column chromatography
- G01N30/62—Detectors specially adapted therefor
- G01N30/72—Mass spectrometers
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- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N30/00—Investigating or analysing materials by separation into components using adsorption, absorption or similar phenomena or using ion-exchange, e.g. chromatography or field flow fractionation
- G01N30/02—Column chromatography
- G01N2030/022—Column chromatography characterised by the kind of separation mechanism
- G01N2030/027—Liquid chromatography
Abstract
The invention provides an establishing method for a high-resolution mass spectrum database of anti-oxidants and antiseptic, and a detection method for anti-oxidants and antiseptic. The detection method comprises the following steps: respectively preparing single-standard methanol solutions of anti-oxidants and antiseptic and carrying out mass spectrometry with a time-of-flight mass spectrometer in a positive ion full scanning mode and a negative ion full scanning mode so as to establish the high-resolution mass spectrum database; and extracting a to-be-detected sample with alcoholic solutions, carrying out mass spectrometry and detecting the anti-oxidants and antiseptic in the sample according to mass spectral data and the high-resolution mass spectrum database. Compared with the prior art, the invention has the advantages that since high-resolution mass spectrum and target analysis strategies are employed for detection of the anti-oxidants and antiseptic, the method can carry out rapid qualitative detection on a plurality of anti-oxidants and antiseptic at the same time without complex pre-treatment steps; a combination of liquid chromatography and high0resolution mass spectrometry is employed, so the method is high in accuracy and sensitivity; and the method can carry out qualitative determination on screened unknown anti-oxidants and antiseptic, so the scope of analysis and monitoring is broadened.
Description
Technical field
The invention belongs to technical field of chemical detection, particularly relate to antioxidant and preservative high resolution mass spectrum
The method for building up of database and detection antioxidant and the method for preservative.
Background technology
2014 Nian Ban China national food safety standard GB 2760 " uses of food additives " and 2011
Year, version compared, and antioxidant list adds tea polyphenol-palmitate and reaches 27 kinds, leaves out in preservative list
4-phenylphenol, 2-phenylphenol sodium salt, ethyl naphthol and sec-butylamine, add epsilon-polylysine and salt
Hydrochlorate, lysozyme, two editions amount to 30 kinds of preservatives.In the face of so many additive, how rapid screening
The focus of multiple additives information, always safety and supervision area.
The carrying out of Tobacco use control makes tobacco consumption that new change to occur, and non-burning class tobacco product, such as nothing
Increasing occur in tobacco products, the sale of novel tobacco goods, and the reality of " Framework Convention on Tobacco Control "
Execute in scheme, then advise the use of additive in tobacco is limited or forbidden, but at present to smokeless cigarette
In grass, the research of additive service condition is less.Additive in above-mentioned China national standard, such as preservative and
The service condition of antioxidant etc., merits attention, and developing screening method fast and accurately is to pacify additive
The premise the most effectively supervised.
Analysis and research about preservative and antioxidant are a lot, during as used comprehensive two dimensional gas chromatography/flight
Between preservative in mass spectrography examination food and antioxidant, use Rtx-5 low-pole column series connection Rtx-17
Polar column orthogonal separation component, determines 32 kinds of objects；Ultra Performance Liquid Chromatography-tandem mass spectrometry measures
Preservative in flavouring and antioxidant, after Solid phase extraction pre-treatment, electron spray quadrupole rod mass spectrum
Carry out multiple-reaction monitoring (MRM) under negative ion mode to analyze, quantitative determine 17 kinds of objects.
GC × GC method resolution ratio is high, peak capacity is big, but data process relatively complicated, are used for more than 100 kinds of groups
Complex matrices analysis is divided to have outstanding advantage, but slightly aobvious inconvenient for rapid screening；QqQ (MRM) spirit
There is some superiority in sensitivity, but in selectivity with analyte quantitative aspects not as high resolution mass spectrum.
HPLC/TOF-MS target analyzes (Target analysis) at numerous areas such as medicine, food, cosmetics
It is widely used.About antioxidant in vegetable oil and the analysis of preservative, method uses anti-phase C18 post,
11 kinds of objects of quantitative analysis under negative ion mode, but for the impact of high resolution mass spectrum examination object
Factor is inquired into the most further.
Further, the most general first mass spectrometric database is more, but can openly obtain two grades of modal data storehouses
Seldom, during especially for some particular analysis monitoring range, if establishing corresponding second order ms database,
The result analyzing object can be confirmed.During known target thing target is analyzed, can be at many chemicals
Matter structural database is searched storehouse and obtains the information such as compound name, molecular formula and structure, as PUBCHEM,
ChemSpider, ChEBI etc., but two grades of modal data information only have less content in MASSBANK.
Summary of the invention
In view of this, the technical problem to be solved in the present invention is to provide antioxidant and preservative high-resolution
The method for building up in mass spectrometric data storehouse and detection antioxidant and the method for preservative, the method pre-treatment is simple,
Quick and precisely, highly sensitive.
The invention provides a kind of method detecting antioxidant and preservative, including:
S1) the list mark methanol solution of antioxidant is prepared；
By the list mark methanol solution of described antioxidant under time of-flight mass spectrometer cation full scan pattern
Carry out mass spectral analysis, the first one-level high resolution mass spectrum data of collection antioxidant and the one or two grade of high-resolution
Mass spectrometric data；
By the list mark methanol solution of described antioxidant under time of-flight mass spectrometer anion full scan pattern
Carry out mass spectral analysis, the second one-level high resolution mass spectrum data of collection antioxidant and the two or two grade of high-resolution
Mass spectrometric data；
The list mark methanol solution of preparation preservative；
The list mark methanol solution of described preservative is entered under time of-flight mass spectrometer cation full scan pattern
Row mass spectral analysis, the first one-level high resolution mass spectrum data of collection preservative and the one or two grade of high resolution mass spectrum
Data；
The list mark methanol solution of described preservative is entered under time of-flight mass spectrometer anion full scan pattern
Row mass spectral analysis, the second one-level high resolution mass spectrum data of collection preservative and the two or two grade of high resolution mass spectrum
Data；
Setting up high resolution mass spectrum database, described high resolution mass spectrum database includes the 1st of antioxidant
Level high resolution mass spectrum data, the one or two grade of high resolution mass spectrum data of antioxidant, the second of antioxidant
One-level high resolution mass spectrum data, the two or two grade of high resolution mass spectrum data of antioxidant, the first of preservative
One-level high resolution mass spectrum data, the one or two grade of high resolution mass spectrum data of preservative, the 2nd 1 of preservative the
Level high resolution mass spectrum data and the two or two grade of high resolution mass spectrum data of preservative；
S2) testing sample alcoholic solution is extracted, obtain extract；
Described extract is carried out mass spectral analysis in time of-flight mass spectrometer cation full scan pattern, obtains
First one-level high resolution mass spectrum data of extract and the one or two grade of high resolution mass spectrum data；
Described extract is carried out mass spectral analysis under time of-flight mass spectrometer anion full scan pattern,
The second one-level high resolution mass spectrum data and the two or two grade of high resolution mass spectrum data to extract；
The first one-level high resolution mass spectrum data according to extract, the one or two grade of high resolution mass spectrum of extract
Data, the second one-level high resolution mass spectrum data of extract, the two or two grade of high resolution mass spectrum number of extract
According to step S1) in high resolution mass spectrum database detection testing sample in antioxidant and preservative.
Preferably, described step S1) particularly as follows:
The list mark methanol solution of preparation antioxidant；
The list mark methanol solution of described antioxidant is carried out liquid-phase chromatographic analysis, obtains the liquid of antioxidant
Phase chromatographic data；
By the list mark methanol solution of described antioxidant under time of-flight mass spectrometer cation full scan pattern
Carry out mass spectral analysis, the first one-level high resolution mass spectrum data of collection antioxidant and the one or two grade of high-resolution
Mass spectrometric data；
By the list mark methanol solution of described antioxidant under time of-flight mass spectrometer anion full scan pattern
Carry out mass spectral analysis, the second one-level high resolution mass spectrum data of collection antioxidant and the two or two grade of high-resolution
Mass spectrometric data；
The list mark methanol solution of preparation preservative；
The list mark methanol solution of described preservative is carried out liquid-phase chromatographic analysis, obtains the liquid phase look of preservative
Modal data；
The list mark methanol solution of described preservative is entered under time of-flight mass spectrometer cation full scan pattern
Row mass spectral analysis, the first one-level high resolution mass spectrum data of collection preservative and the one or two grade of high resolution mass spectrum
Data；
The list mark methanol solution of described preservative is entered under time of-flight mass spectrometer anion full scan pattern
Row mass spectral analysis, the second one-level high resolution mass spectrum data of collection preservative and the two or two grade of high resolution mass spectrum
Data；
Setting up high resolution mass spectrum database, described mass spectrometric data storehouse includes the first one-level high score of antioxidant
Distinguish the second one-level height of mass spectrometric data, the one or two grade of high resolution mass spectrum data of antioxidant, antioxidant
Resolution mass spectra, the two or two grade of high resolution mass spectrum data of antioxidant, the first one-level height of preservative
Resolution mass spectra, the one or two grade of high resolution mass spectrum data of preservative, the second one-level high score of preservative
Distinguish the two or two grade of high resolution mass spectrum data of mass spectrometric data and preservative.
Preferably, described step S2) particularly as follows:
Testing sample alcoholic solution is extracted, obtains extract；
Described extract is first carried out liquid-phase chromatographic analysis, obtains the Liquid Chromatography data of extract；
Extract through liquid-phase chromatographic analysis is carried out in time of-flight mass spectrometer cation full scan pattern
Mass spectral analysis, obtains the first one-level high resolution mass spectrum data of extract and the one or two grade of high resolution mass spectrum number
According to；
Extract through liquid-phase chromatographic analysis is entered under time of-flight mass spectrometer anion full scan pattern
Row mass spectral analysis, obtains the second one-level high resolution mass spectrum data of extract and the two or two grade of high resolution mass spectrum
Data；
Liquid Chromatography data according to extract, the first one-level high resolution mass spectrum data of extract, extraction
One or two grade of high resolution mass spectrum data of liquid, the second one-level high resolution mass spectrum data of extract, extract
The two or two grade of high resolution mass spectrum data and step S1) in high resolution mass spectrum database detection testing sample
In antioxidant and preservative.
Preferably, the mobile phase A of described liquid-phase chromatographic analysis is formic acid water, and Mobile phase B is methyl alcohol and first
The mixed liquor of acid, the Mobile phase B isocratic elution of 70%.
Preferably, the Mass Spectrometry Conditions of described cation full scan pattern is: electron spray ESI ion gun；Hair
Tubule voltage 3000～4000V；Atomization gas 1～3Bar；It is dried temperature 150 DEG C～200 DEG C；It is dried gas
Flow velocity 2～10l/min；Mass scan range m/z 50～1000amu；Six grades of bar radio-frequency voltages 150.0～300.0
Vpp；Collision cell radio-frequency voltage 500.0～800.0Vpp.
Preferably, the Mass Spectrometry Conditions of described anion full scan pattern is: electron spray ESI ion gun；Hair
Tubule voltage 2500～3500V；Atomization gas 1～3Bar；It is dried temperature 150 DEG C～200 DEG C；It is dried gas
Flow velocity 2～10l/min；Mass scan range m/z 50～1000amu；Level Four bar ion energy 2.0～6.0eV；
Collision cell penetrates energy 5.0～10.0eV；Collision cell radio-frequency voltage 500.0～800.0Vpp；The transmission time
80.0～120.0 μ s；Prefocus frequency 3.0～7.0 μ s.
Preferably, described step S1) in one-level high resolution mass spectrum data and two grades of high resolution mass spectrum data
Accurate mass number error is less than or equal to 2mDa, and isotope peak shape deviation is less than or equal to 20.
Preferably, described antioxidant and preservative are thiabendazole, ethoxyquinoline, glycyrrhizic acid, right
Coumaric acid, propylgallate, natamycin, methyl hydroxybenzoate, ethylparaben, para hydroxybenzene first
Propyl propionate, n Heptyl p hydroxybenzoate, p-phenyl phenol, benzyl p-hydroxybenzoate, 4-HBA
Butyl ester, 4-HBA isobutyl ester, 4-hexyl resorcin, 2,4-D are sour, P-hydroxybenzoic acid is the most pungent
Ester, forulic acid, TBHQ, 3,3-dithiodipropionic acid dilauryl ester, Rosmarinic acid are with anti-
Bad hematic acid.
Preferably, described testing sample is smokeless tobacco.
Present invention also offers a kind of antioxidant or the method for building up of preservative high resolution mass spectrum database,
Including:
The list mark methanol solution of preparation antioxidant；
By the list mark methanol solution of described antioxidant under time of-flight mass spectrometer cation full scan pattern
Carry out mass spectral analysis, the first one-level high resolution mass spectrum data of collection antioxidant and the one or two grade of high-resolution
Mass spectrometric data；
By the list mark methanol solution of described antioxidant under time of-flight mass spectrometer anion full scan pattern
Carry out mass spectral analysis, the second one-level high resolution mass spectrum data of collection antioxidant and the two or two grade of high-resolution
Mass spectrometric data；
The list mark methanol solution of preparation preservative；
The list mark methanol solution of described preservative is entered under time of-flight mass spectrometer cation full scan pattern
Row mass spectral analysis, the first one-level high resolution mass spectrum data of collection preservative and the one or two grade of high resolution mass spectrum
Data；
The list mark methanol solution of described preservative is entered under time of-flight mass spectrometer anion full scan pattern
Row mass spectral analysis, gathers the second one-level high resolution mass spectrum data and the two or the two high-resolution level mass spectrum of preservative
Data；
Setting up high resolution mass spectrum database, described high resolution mass spectrum database includes the 1st of antioxidant
Level high resolution mass spectrum data, the one or two grade of high resolution mass spectrum data of antioxidant, the second of antioxidant
One-level high resolution mass spectrum data, the two or two grade of high resolution mass spectrum data of antioxidant, the first of preservative
One-level high resolution mass spectrum data, the one or two grade of high resolution mass spectrum data of preservative, the 2nd 1 of preservative the
Level high resolution mass spectrum data and the two or two grade of high resolution mass spectrum data of preservative.
The invention provides antioxidant and the method for building up of preservative high resolution mass spectrum database and detection is anti-
Oxidant and the method for preservative, including: S1) prepare the list mark methanol solution of antioxidant；By described
The list mark methanol solution of antioxidant carries out mass spectrum under time of-flight mass spectrometer cation full scan pattern and divides
Analysis, the first one-level high resolution mass spectrum data of collection antioxidant and the one or two grade of high resolution mass spectrum data；
The list mark methanol solution of described antioxidant is carried out under time of-flight mass spectrometer anion full scan pattern
Mass spectral analysis, the second one-level high resolution mass spectrum data of collection antioxidant and the two or two grade of high resolution mass spectrum
Data；The list mark methanol solution of preparation preservative；By the list mark methanol solution of described preservative when flight
Between carry out mass spectral analysis under mass spectrograph cation full scan pattern, gather the first one-level high-resolution of preservative
Mass spectrometric data and the one or two grade of high resolution mass spectrum data；By the list mark methanol solution of described preservative in flight
Carry out mass spectral analysis under time mass spectrum instrument anion full scan pattern, gather the second one-level high score of preservative
Distinguish mass spectrometric data and the two or two grade of high resolution mass spectrum data；Set up high resolution mass spectrum database, described high score
Distinguish mass spectrometric data storehouse include the first one-level high resolution mass spectrum data of antioxidant, the 1st of antioxidant the
Level high resolution mass spectrum data, the second one-level high resolution mass spectrum data of antioxidant, the second of antioxidant
Two grades of high resolution mass spectrum data, the first one-level high resolution mass spectrum data of preservative, the 1st of preservative the
Level high resolution mass spectrum data, the second one-level high resolution mass spectrum data of preservative and the two or two grade of preservative
High resolution mass spectrum data；S2) testing sample alcoholic solution is extracted, obtain extract；By described extraction
Liquid carries out mass spectral analysis in time of-flight mass spectrometer cation full scan pattern, obtains the 1st of extract the
Level high resolution mass spectrum data and the one or two grade of high resolution mass spectrum data；By described extract in flight time matter
Carry out mass spectral analysis under spectrometer anion full scan pattern, obtain the second one-level high resolution mass spectrum of extract
Data and the two or two grade of high resolution mass spectrum data；The first one-level high resolution mass spectrum data according to extract,
One or two grade of high resolution mass spectrum data of extract, the second one-level high resolution mass spectrum data of extract, carry
Take the two or two grade of high resolution mass spectrum data and step S1 of liquid) in high resolution mass spectrum database detection to be measured
Antioxidant in sample and preservative.Compared with prior art, the present invention uses high resolution mass spectrum and target
Antioxidant and preservative are detected by mark analysis strategy, and it need not the pre-treatment step of complexity, i.e.
Multiple antioxidant and preservative can be carried out fast qualitative detection simultaneously；Further use liquid chromatogram
With high resolution mass spectrum associated with detection method not only accuracy high, highly sensitive, also examination can be gone out not
Know that antioxidant and preservative qualitatively judge, expand the scope of research and application.
Accompanying drawing explanation
Valve connecting mode schematic diagram when Fig. 1 is time of-flight mass spectrometer of the present invention employing direct mass spectrum sample introduction；
Fig. 2 is the one-level high resolution mass spectrum database example figure of Self-built Database in the embodiment of the present invention 1；
Fig. 3 is two grades of high resolution mass spectrum database example figures of Self-built Database in the embodiment of the present invention 1；
Fig. 4 is the second order ms figure obtaining unknown material in the embodiment of the present invention 2 under Auto MS/MS pattern.
Detailed description of the invention
Below in conjunction with the accompanying drawing of the embodiment of the present invention, the technical scheme in the embodiment of the present invention is carried out clearly
Chu, it is fully described by, it is clear that described embodiment is only a part of embodiment of the present invention, and not
It it is whole embodiments.Based on the embodiment in the present invention, those of ordinary skill in the art are not making
The every other embodiment obtained under creative work premise, broadly falls into the scope of protection of the invention.
The invention provides a kind of antioxidant and the method for building up of preservative high resolution mass spectrum database, bag
Include:
The list mark methanol solution of preparation antioxidant；
By the list mark methanol solution of described antioxidant under time of-flight mass spectrometer cation full scan pattern
Carry out mass spectral analysis, the first one-level high resolution mass spectrum data of collection antioxidant and the one or two grade of high-resolution
Mass spectrometric data；
By the list mark methanol solution of described antioxidant under time of-flight mass spectrometer anion full scan pattern
Carry out mass spectral analysis, the second one-level high resolution mass spectrum data of collection antioxidant and the two or two grade of high-resolution
Mass spectrometric data；
The list mark methanol solution of preparation preservative；
The list mark methanol solution of described preservative is entered under time of-flight mass spectrometer cation full scan pattern
Row mass spectral analysis, the first one-level high resolution mass spectrum data of collection preservative and the one or two grade of high resolution mass spectrum
Data；
The list mark methanol solution of described preservative is entered under time of-flight mass spectrometer anion full scan pattern
Row mass spectral analysis, the second one-level high resolution mass spectrum data of collection preservative and the two or two grade of high resolution mass spectrum
Data；
Setting up high resolution mass spectrum database, described high resolution mass spectrum database includes the 1st of antioxidant
Level high resolution mass spectrum data, the one or two grade of high resolution mass spectrum data of antioxidant, the second of antioxidant
One-level high resolution mass spectrum data, the two or two grade of high resolution mass spectrum data of antioxidant, the first of preservative
One-level high resolution mass spectrum data, the one or two grade of high resolution mass spectrum data of preservative, the 2nd 1 of preservative the
Level high resolution mass spectrum data and the two or two grade of high resolution mass spectrum data of preservative.
Wherein, described antioxidant and preservative be preferably thiabendazole, ethoxyquinoline, glycyrrhizic acid,
P-Coumaric Acid, propylgallate, natamycin, methyl hydroxybenzoate, ethylparaben, para hydroxybenzene
Propyl formate, n Heptyl p hydroxybenzoate, p-phenyl phenol, benzyl p-hydroxybenzoate, 4-hydroxy benzenes first
Acid butyl ester, 4-HBA isobutyl ester, 4-hexyl resorcin, 2,4-D acid, P-hydroxybenzoic acid are just
Monooctyl ester, forulic acid, TBHQ, 3,3-dithiodipropionic acid dilauryl ester, Rosmarinic acid with
Ascorbic acid, its English name and molecular formula are shown in Table 1.
Table 1 antioxidant and the title of preservative and molecular formula
The present invention prepares the list mark methanol solution of antioxidant the most in accordance with the following methods: weigh respectively
The standard items of 7.5～10.0mg antioxidants, with in methanol constant volume to 25ml volumetric flask, are configured to 0.3～0.4
The list mark solution of mg/ml；Pipette the list mark solution of 25～35 μ l the most respectively, by methanol constant volume to 10ml
In volumetric flask, it is configured to the list mark methanol solution of 1.0 μ g/ml.
By the list mark methanol solution of described antioxidant under time of-flight mass spectrometer cation full scan pattern
Carry out mass spectral analysis, the first one-level high resolution mass spectrum data of collection antioxidant and the one or two grade of high-resolution
Mass spectrometric data；
By the list mark methanol solution of described antioxidant under time of-flight mass spectrometer anion full scan pattern
Carry out mass spectral analysis, the second one-level high resolution mass spectrum data of collection antioxidant and the two or two grade of high-resolution
Mass spectrometric data；
Antioxidant list mark methanol solution of the present invention uses direct mass spectrum sample introduction, now valve connecting mode such as Fig. 1
Shown in, the list mark methanol solution of antioxidant and preservative forwards to couple port5 from mass spectrum atomization chamber injection port,
The HPLC disconnecting port3 couples, and is introduced by pin pump from atomization chamber injection port by single mark methanol solution；Institute
The flow velocity stating pin pump is preferably set to 100～200 μ l/h, more preferably 150～200 μ l/h, is further preferably
160～190 μ l/h, most preferably 180 μ l/h.
Wherein, the Mass Spectrometry Conditions of described time of-flight mass spectrometer cation full scan pattern is preferably: EFI
Mist ESI ion gun；Capillary voltage 3000～4000V；Atomization gas 1～3Bar；It is dried temperature 150 DEG C
～200 DEG C；Dry gas stream speed 2～10l/min；Mass scan range m/z 50～1000amu；Six grades of bars are penetrated
Frequently voltage 150.0～300.0Vpp；Collision cell radio-frequency voltage 500.0～800.0Vpp；More preferably: EFI
Mist ESI ion gun；Capillary voltage 3500V；Atomization gas 1.5Bar；It is dried temperature 180 DEG C；Dry
Pathogenic dryness flow velocity 6.0l/min；Mass scan range m/z 50～1000amu；Six grades of bar radio-frequency voltages 200.0
Vpp；Collision cell radio-frequency voltage 600.0Vpp.
The Mass Spectrometry Conditions of described time of-flight mass spectrometer anion full scan pattern is preferably: electron spray ESI
Ion gun；Capillary voltage 2500～3500V；Atomization gas 1～3Bar；It is dried temperature 150 DEG C～200 DEG C；
Dry gas stream speed 2～10l/min；Mass scan range m/z 50～1000amu；Level Four bar ion energy
2.0～6.0eV；Collision cell penetrates energy 5.0～10.0eV；Collision cell radio-frequency voltage 500.0～800.0Vpp；
Transmission time 80.0～120.0 μ s；Prefocus frequency 3.0～7.0 μ s.；More preferably: electron spray ESI from
Component；Capillary voltage 3000V；Atomization gas 1.5Bar；It is dried temperature 180 DEG C；Dry gas stream speed
6.0l/min；Mass scan range m/z 50～1000amu；Level Four bar ion energy 4.0eV；Collision cell
Penetrate energy 7.0eV；Collision cell radio-frequency voltage 600.0Vpp；Transmission time 100.0 μ s；Prefocus frequency
5.0μs。
For eliminating the solvent impact on mass spectrometric data, preferably entirely sweep in cation full scan pattern and anion
Retouch and under pattern, all use sodium methoxide solution to correct mass spectrograph.The concentration of described sodium methoxide is preferably 1～50
Mmol/L, more preferably 5～30mmol/L, be further preferably 10～20mmol/L, most preferably 10
mmol/L。
First one-level high resolution mass spectrum data of collection antioxidant and the one or two grade of high resolution mass spectrum data,
Preferably accurate mass number error is less than or equal to 2mDa, and isotope peak shape deviation is less than or equal to 20；Gather anti-
Second one-level high resolution mass spectrum data of oxidant and the two or two grade of high resolution mass spectrum data, preferably accurately matter
Amount number error is less than or equal to 20 less than or equal to 2mDa, isotope peak shape deviation.
The list mark methanol solution of preparation preservative；Described compound method and the list mark methanol solution of antioxidant
Compound method identical, do not repeat them here.
The list mark methanol solution of described preservative is entered under time of-flight mass spectrometer cation full scan pattern
Row mass spectral analysis, the first one-level high resolution mass spectrum data of collection preservative and the one or two grade of high resolution mass spectrum
Data；By the list mark methanol solution of described preservative under time of-flight mass spectrometer anion full scan pattern
Carry out mass spectral analysis, the second one-level high resolution mass spectrum data of collection preservative and the two or two grade of high-resolution matter
Modal data；Described time of-flight mass spectrometer cation full scan pattern is with anion full scan pattern the most ibid
Described, do not repeat them here.
Setting up high resolution mass spectrum database, described high resolution mass spectrum database includes the 1st of antioxidant
Level high resolution mass spectrum data, the one or two grade of high resolution mass spectrum data of antioxidant, the second of antioxidant
One-level high resolution mass spectrum data, the two or two grade of high resolution mass spectrum data of antioxidant, the first of preservative
One-level high resolution mass spectrum data, the one or two grade of high resolution mass spectrum data of preservative, the 2nd 1 of preservative the
Level high resolution mass spectrum data and the two or two grade of high resolution mass spectrum data of preservative.The present invention is preferably by antioxygen
The one-level high resolution mass spectrum data of agent and preservative import Library with two grades of high resolution mass spectrum data
Editor software, sets up antioxidant and preservative high resolution mass spectrum database.
Heretofore described high resolution mass spectrum database preferably include antioxidant and the title of preservative,
No. CAS, chemical formula, chemical constitution, molecular weight, first mass spectrometric data, second order ms data, target
Thing impact energy, the accurate mass number of fragments characteristic ion and precision.
Antioxidant and preservative high resolution mass spectrum database that the present invention sets up can provide antioxidant and prevent
The rotten compound essential information of agent, the parameter of first mass spectrometric data and the parameter of second order ms data.
Present invention also offers one and utilize above-mentioned antioxidant and the detection of preservative high resolution mass spectrum database
Antioxidant and the method for preservative, including:
S1) the list mark methanol solution of antioxidant is prepared；
By the list mark methanol solution of described antioxidant under time of-flight mass spectrometer cation full scan pattern
Carry out mass spectral analysis, the first one-level high resolution mass spectrum data of collection antioxidant and the one or two grade of high-resolution
Mass spectrometric data；
By the list mark methanol solution of described antioxidant under time of-flight mass spectrometer anion full scan pattern
Carry out mass spectral analysis, the second one-level high resolution mass spectrum data of collection antioxidant and the two or two grade of high-resolution
Mass spectrometric data；
The list mark methanol solution of preparation preservative；
The list mark methanol solution of described preservative is entered under time of-flight mass spectrometer cation full scan pattern
Row mass spectral analysis, the first one-level high resolution mass spectrum data of collection preservative and the one or two grade of high resolution mass spectrum
Data；
The list mark methanol solution of described preservative is entered under time of-flight mass spectrometer anion full scan pattern
Row mass spectral analysis, the second one-level high resolution mass spectrum data of collection preservative and the two or two grade of high resolution mass spectrum
Data；
Setting up high resolution mass spectrum database, described high resolution mass spectrum database includes the 1st of antioxidant
Level high resolution mass spectrum data, the one or two grade of high resolution mass spectrum data of antioxidant, the second of antioxidant
One-level high resolution mass spectrum data, the two or two grade of high resolution mass spectrum data of antioxidant, the first of preservative
One-level high resolution mass spectrum data, the one or two grade of high resolution mass spectrum data of preservative, the 2nd 1 of preservative the
Level high resolution mass spectrum data and the two or two grade of high resolution mass spectrum data of preservative；
S2) testing sample alcoholic solution is extracted, obtain extract；
Described extract is carried out mass spectral analysis in time of-flight mass spectrometer cation full scan pattern, obtains
First one-level high resolution mass spectrum data of extract and the one or two grade of high resolution mass spectrum data；
Described extract is carried out mass spectral analysis under time of-flight mass spectrometer anion full scan pattern,
The second one-level high resolution mass spectrum data and the two or two grade of high resolution mass spectrum data to extract；
The first one-level high resolution mass spectrum data according to extract, the one or two grade of high resolution mass spectrum of extract
Data, the second one-level high resolution mass spectrum data of extract, the two or two grade of high resolution mass spectrum number of extract
According to step S1) in high resolution mass spectrum database detection testing sample in antioxidant and preservative.
Wherein, described step S1) in set up antioxidant and the method for preservative high resolution mass spectrum database
Same as above, do not repeat them here.
In the present invention, in order to improve screening speed and the efficiency of antioxidant and preservative, institute further
State S1) in the most also include liquid-phase chromatographic analysis, first by the list mark methanol solution of antioxidant or preservative
Carrying out liquid-phase chromatographic analysis, then carry out mass spectral analysis, i.e. LC-MS, now, described step S1 is preferred
Particularly as follows:
The list mark methanol solution of preparation antioxidant；
The list mark methanol solution of described antioxidant is carried out liquid-phase chromatographic analysis, obtains the liquid of antioxidant
Phase chromatographic data；
By the list mark methanol solution of described antioxidant under time of-flight mass spectrometer cation full scan pattern
Carry out mass spectral analysis, the first one-level high resolution mass spectrum data of collection antioxidant and the one or two grade of high-resolution
Mass spectrometric data；
By the list mark methanol solution of described antioxidant under time of-flight mass spectrometer anion full scan pattern
Carry out mass spectral analysis, the second one-level high resolution mass spectrum data of collection antioxidant and the two or two grade of high-resolution
Mass spectrometric data；
The list mark methanol solution of preparation preservative；
The list mark methanol solution of described preservative is carried out liquid-phase chromatographic analysis, obtains the liquid phase look of preservative
Modal data；
The list mark methanol solution of described preservative is entered under time of-flight mass spectrometer cation full scan pattern
Row mass spectral analysis, the first one-level high resolution mass spectrum data of collection preservative and the one or two grade of high resolution mass spectrum
Data；
The list mark methanol solution of described preservative is entered under time of-flight mass spectrometer anion full scan pattern
Row mass spectral analysis, the second one-level high resolution mass spectrum data of collection preservative and the two or two grade of high resolution mass spectrum
Data；
Setting up high resolution mass spectrum database, described high resolution mass spectrum database includes the 1st of antioxidant
Level high resolution mass spectrum data, the one or two grade of high resolution mass spectrum data of antioxidant, the second of antioxidant
One-level high resolution mass spectrum data, the two or two grade of high resolution mass spectrum data of antioxidant, the first of preservative
One-level high resolution mass spectrum data, the one or two grade of high resolution mass spectrum data of preservative, the 2nd 1 of preservative the
Level high resolution mass spectrum data and the two or two grade of high resolution mass spectrum data of preservative.
Wherein, the kind of described antioxidant and preservative is same as above, does not repeats them here, in detection
In antioxidant more preferably the most as shown in table 2 and preservative.
Table 2 antioxidant and the title of preservative and molecular formula
The liquid chromatogram that described liquid chromatogram is well known to those skilled in the art, there is no special restriction,
The present invention is preferably high performance liquid chromatography or Ultra Performance Liquid Chromatography；The flowing of described liquid-phase chromatographic analysis
Phase A is formic acid water, and Mobile phase B is the mixed liquor of methyl alcohol and formic acid, the Mobile phase B isocratic elution of 70%；
Wherein, the pH value of described formic acid water is preferably 2～3, and more preferably 2.5～3, be further preferably 2.7；Described
In mixed liquor, the volume content of formic acid is preferably 0.1%～0.3%, more preferably 0.1%～0.2%, further preferably
It is 0.1%～0.15%, most preferably 0.1%；Chromatographic column used by described liquid-phase chromatographic analysis is preferably Luna
C8 liquid-phase chromatographic column；The specification of described chromatographic column is preferably 4.6mm × 3 μ m 150mm；Described chromatogram
The temperature of post is preferably 30 DEG C～40 DEG C, more preferably 32 DEG C～38 DEG C, is further preferably 34 DEG C～36 DEG C,
It is preferably 35 DEG C；The sampling volume of described liquid chromatogram is preferably 3～10 μ l, more preferably 5～10 μ l,
It is further preferably 5～8 μ l, most preferably 5 μ l.
The condition of described mass spectral analysis is same as above, does not repeats them here.
Testing sample alcoholic solution is extracted, obtains extract；Described testing sample is people in the art
Testing sample known to Yuan, there is no special restriction, is preferably smokeless tobacco in the present invention；Described
Alcoholic solution is preferably the mixed liquor of methyl alcohol and water；Described methyl alcohol is preferably (1～5) with the volume ratio of water: 1,
More preferably (2～4): 1, most preferably 3:1；Described testing sample is preferably 0.1 with the ratio of alcoholic solution
G:(1～3) ml, more preferably 0.1g:2ml；Being preferably a step of described extraction: by testing sample
Mix with alcoholic solution and carry out ultrasonic, vortex, centrifugal, filtration treatment successively, obtain extract；Described super
The time of sound is preferably 5～15min, and more preferably 8～12min, be further preferably 10min；Described vortex
Time be preferably 10～20min, more preferably 13～18min, be further preferably 15min；Described centrifugal
Speed be preferably 5000～20000r/min, more preferably 8000～15000r/min, be further preferably
10000～12000r/min, most preferably 10000r/min；The described centrifugal time is preferably 5～20min,
More preferably 8～15min, it is further preferably 10min；Described filtration preferably employs 0.1～0.5 μm filter membrane and enters
Row filters, and more preferably uses 0.2 μm filter membrane to filter.
According to the present invention, the most first extract is carried out liquid-phase chromatographic analysis, obtain the liquid phase look of extract
Modal data；By the extract through liquid-phase chromatographic analysis in time of-flight mass spectrometer cation full scan pattern
Carry out mass spectral analysis, obtain the first one-level high resolution mass spectrum data of extract and the one or two grade of high-resolution matter
Modal data；By the extract through liquid-phase chromatographic analysis in time of-flight mass spectrometer anion full scan pattern
Under carry out mass spectral analysis, obtain the second one-level high resolution mass spectrum data of extract and the two or two grade of high-resolution
Mass spectrometric data；
Wherein, the condition of described liquid chromatogram is the most same as above with described mass spectrographic condition, the most superfluous at this
State；In order to reduce the solute impact on chromatographic data, it is preferred to use sodium methoxide solution correction mass spectrograph；Institute
The concentration stating sodium methoxide is preferably 1～50mmol/L, and more preferably 5～30mmol/L, be further preferably 10～20
Mmol/L, most preferably 10mmol/L.
Liquid Chromatography data according to extract, the first one-level high resolution mass spectrum data of extract, extraction
One or two grade of high resolution mass spectrum data of liquid, the second one-level high resolution mass spectrum data of extract, extract
The two or two grade of high resolution mass spectrum data and step S1) in high resolution mass spectrum database detection testing sample
In antioxidant and preservative.
The present invention need not the pre-treatment step of complexity, can enter multiple antioxidant and preservative simultaneously
Row fast qualitative detects；The detection method not only accuracy that the present invention provides simultaneously is high, highly sensitive, also
The unknown antioxidant detected and preservative can be qualitatively judged, expand the scope of research and application；
The inventive method uses high resolution mass spectrum to gather exact mass number, and the resolution ratio of antioxidant and preservative can
Reach 2～30,000.
In order to further illustrate the present invention, the antioxidant that the present invention provided below in conjunction with embodiment and anti-
The method for building up of rotten agent high resolution mass spectrum database and the method for detection antioxidant and preservative are carried out in detail
Describe.
Reagent used in following example is commercially available.
Embodiment 1
1.1 standard liquid preparations
Standard liquid: weigh 7.5-10.0mg standard items respectively, with in methanol constant volume to 25mL volumetric flask,
It is configured to the list mark methanol solution of 0.3-0.4mg/mL；Pipette the list mark methanol solution of 25-35 μ L respectively,
With in methanol constant volume to 10mL volumetric flask, being configured to the list mark methanol solution of about 1.0 μ g/mL, 4 DEG C cold
Hide stand-by.
1.2 Mass Spectrometry Conditions
Electron spray ESI ion gun: cation full scan pattern, capillary voltage: 3500V；Atomization gas:
1.5Bar；Dry temperature: 180 DEG C；Dry gas stream speed: 6.0l/min；Mass scan range: m/z 50-1000
amu；Six grades of bar radio-frequency voltage: 200.0Vpp；Collision cell radio-frequency voltage: 600.0Vpp.
Anion full scan pattern；Capillary voltage: 3000V；Atomization gas: 1.5Bar；It is dried temperature
Degree: 180 DEG C；Dry gas stream speed: 6.0l/min；Mass scan range: m/z 50-1000amu；Quadrupole
Bar ion energy: 4.0eV；Collision cell energy: 7.0eV；Collision cell radio-frequency voltage: 600.0Vpp；Pass
The defeated time: 100.0 μ s；Prefocus frequency: 5.0 μ s.
All using the sodium formate solution of 10mmol/L as mass number correcting fluid under negative ions pattern.
When 1.3 mass spectrographs use direct mass spectrum sample introduction, valve connecting mode is as it is shown in figure 1, standard correction liquid is from matter
Spectrum atomization chamber injection port forwards to couple port 5, and the HPLC disconnecting port 3 couples, by single mark methanol solution
Being introduced by pin pump (syringe pump) from atomization chamber injection port, pin flow rate pump is set to 180 μ L/h.
1.4, with the sodium formate solution correction mass spectrograph of 10mmol/L, gather antioxidant and the one of preservative
Level, second order ms data, make accurate mass number error≤2mDa, isotope peak shape deviation≤20.
1.5 open the Library self-built new database of Editor software, import the antioxidant and anticorrosion collected
The one-level of agent, second order ms data.The one-level of Self-built Database, two grades of high resolution mass spectrum data instance are such as
Shown in Fig. 2, Fig. 3.
Embodiment 2
2.1 standard liquid preparations
Standard reserving solution: weigh 7.5～10.0mg standard items respectively, by methanol constant volume to 25mL volumetric flask
In, it is configured to the list mark methanol solution of 0.3-0.4mg/mL；The list mark methyl alcohol pipetting 25-35 μ L respectively is molten
Liquid, with in methanol constant volume to 10mL volumetric flask, is configured to the list mark methanol solution of about 1.0 μ g/mL, 4 DEG C
Refrigerate stand-by.
Single mark working solution: the list mark of 1.0 μ g/mL is diluted respectively 1 times (the mono-mark of about 500ppb)；
Mixed mark working solution: pipette the list mark methanol solution of 15-20 μ L 0.3-0.4mg/mL respectively, to 10mL
In volumetric flask, with methanol constant volume (the mixed mark of about 500ppb).4 DEG C of refrigerations are stand-by.
2.2 chromatographic condition
High-efficient liquid phase chromatogram condition: Luna C8 liquid-phase chromatographic column (4.6mm × 3 μ m 150mm,
Phenomenex company).
Mobile phase A is formic acid water (pH 2.7), and Mobile phase B is methyl alcohol (0.1% formic acid)；Sample introduction body
Amassing is 5 μ L；Column temperature is 35 DEG C；Flow velocity is 400 μ L/min；70%B isocratic elution.
2.3 Mass Spectrometry Conditions
Electron spray ESI ion gun: cation full scan pattern, capillary voltage: 3500V；Atomization gas:
1.5Bar；Dry temperature: 180 DEG C；Dry gas stream speed: 6.0l/min；Mass scan range: m/z 50-1000
amu；Six grades of bar radio-frequency voltage: 200.0Vpp；Collision cell radio-frequency voltage: 600.0Vpp.
Anion full scan pattern；Capillary voltage: 3000V；Atomization gas: 1.5Bar；It is dried temperature
Degree: 180 DEG C；Dry gas stream speed: 6.0l/min；Mass scan range: m/z 50-1000amu；Quadrupole
Bar ion energy: 4.0eV；Collision cell energy: 7.0eV；Collision cell radio-frequency voltage: 600.0Vpp；Pass
The defeated time: 100.0 μ s；Prefocus frequency: 5.0 μ s.
All using the sodium formate solution of 10mmol/L as mass number correcting fluid under negative ions pattern.
2.4 sample treatment
Weigh five parts of 0.1g (being accurate to 0.01g) smokeless tobacco samples in centrifuge tube, add 2mL first
Alcohol: the aqueous solution (volume ratio 3:1), ultrasonic 10min, vortex 15min, 10000r/min be centrifuged 10min,
Take supernatant and cross 0.2 μm filter membrane, obtain extract.
Every part takes 200 μ L mixing and mixes mark as tobacco extraction liquid matrix, addition, and concentration is 250ppb.
2.5 set up high resolution mass spectrum examination database
Obtaining chromatogram and one-level, second order ms data with single mark methanol solution, data prediction uses Bruker
The Data Analysis software of company obtains object accurate mass number and retention time, Smart Formula
Manually identifies chromatographic peak, uses Target Analysis software to set up database, and database mainly includes
The projects such as the sub-mass-to-charge ratio of object quasi-molecular ion peak, retention time, molecular formula, object English name.
2.6 data process
Data prediction uses the Data Analysis software of Bruker company to obtain object accurate mass number
And retention time, Smart Formula manually identifies chromatographic peak, uses self-built high resolution mass spectrum examination
Database qualitatively screening target compound, mass number error range is ± 5.0mDa, isotope peak shape deviation
Decision content≤20.
The result of 2.7 known multiple target objects rapid screenings
For the known substance of Target Analysis examination hit, this experiment is at negative ion mode, cation mould
The screening results of the lower 18 kinds of objects of formula is as shown in table 2, mass number error all≤5.0mDa, isotope
Peak shape deviation is respectively less than 20, shift of retention time≤0.15min.And the ion of the compound that score value is 2
Respond the most relatively low.
Influence factor about Target Analysis: under identical chromatographic conditions, the retention time drift of compound
Move the least；Analyze smokeless tobacco extraction solution and add mixed mark solution and the object of mixed mark methanol solution
Examination, examination score value is identical, illustrates that screening results is had little to no effect by matrix；At HPLC and UPLC
Under analysis condition, resolution ratio and the sensitivity of object there are differences, and under the conditions of HPLC, object separates
Degree height, and under the conditions of UPLC, response increases the ion examination being conducive to content low, high resolution mass spectrum
Advantage make to obtain preferable screening results at two kinds of chromatographic conditions；And the accuracy of mass number is shadow
Ring the key factor of screening results, on pretreatment, need time of-flight mass spectrometer is carried out mass number correction,
In an experiment, each sample introduction all uses sodium formate to correct, and has met mass number error requirements, improves examination
Hit rate.
Table 3 antioxidant and preservative target analysis result (* is isomer)
The unknown material that the 2.8 pairs of examinations go out carries out qualitative
The unknown material arrived for Target Analysis examination, its molecular formula measurable, carries out second order ms figure
Confirmation, to identify unknown material.As in tobacco substrate assay, under negative ion mode, set mass number by mistake
Difference≤5.0mDa, mSigma value is less than 50, retrieves unknown material ion m/z=191.0544, it was predicted that should
Unknown material molecular formula is shown in Table 4.
The molecular formula of the unknown material quasi-molecular ion of table 4 prediction
Molecular formula C is retrieved in PUBCHEM Chemicals Database7H12O6, retrieval result is chinic acid,
Chinic acid is antioxidant and antisepsis antistaling agent, can be used as fragrance-enhancing tobacco agent.
C8H8N4O2Retrieval result be 4-methyl-6-nitro-1,4-dihydro-1,2,4-benzotriazine；
Other do not retrieve compound name.
The second order ms figure of this unknown material is obtained as shown in Figure 4, the mass spectrum recorded under Auto MS/MS pattern
Data can be retrieved in existing mass spectrometric data storehouse (such as Mass Bank), with D-(-)-Quinic acid
Mass number difference be shown in Table 5, this database has the ESI-QqTOF-MS data of more than 800 compound,
Document claims its data source non-standard unified analysis condition.
Difference between table 5 unknown material experiment value and the second order ms data of database retrieval
Claims (10)
1. the method detecting antioxidant and preservative, it is characterised in that including:
S1) the list mark methanol solution of antioxidant is prepared；
By the list mark methanol solution of described antioxidant under time of-flight mass spectrometer cation full scan pattern
Carry out mass spectral analysis, the first one-level high resolution mass spectrum data of collection antioxidant and the one or two grade of high-resolution
Mass spectrometric data；
By the list mark methanol solution of described antioxidant under time of-flight mass spectrometer anion full scan pattern
Carry out mass spectral analysis, the second one-level high resolution mass spectrum data of collection antioxidant and the two or two grade of high-resolution
Mass spectrometric data；
The list mark methanol solution of preparation preservative；
The list mark methanol solution of described preservative is entered under time of-flight mass spectrometer cation full scan pattern
Row mass spectral analysis, the first one-level high resolution mass spectrum data of collection preservative and the one or two grade of high resolution mass spectrum
Data；
The list mark methanol solution of described preservative is entered under time of-flight mass spectrometer anion full scan pattern
Row mass spectral analysis, the second one-level high resolution mass spectrum data of collection preservative and the two or two grade of high resolution mass spectrum
Data；
Setting up high resolution mass spectrum database, described high resolution mass spectrum database includes the 1st of antioxidant
Level high resolution mass spectrum data, the one or two grade of high resolution mass spectrum data of antioxidant, the second of antioxidant
One-level high resolution mass spectrum data, the two or two grade of high resolution mass spectrum data of antioxidant, the first of preservative
One-level high resolution mass spectrum data, the one or two grade of high resolution mass spectrum data of preservative, the 2nd 1 of preservative the
Level high resolution mass spectrum data and the two or two grade of high resolution mass spectrum data of preservative；
S2) testing sample alcoholic solution is extracted, obtain extract；
Described extract is carried out mass spectral analysis in time of-flight mass spectrometer cation full scan pattern, obtains
First one-level high resolution mass spectrum data of extract and the one or two grade of high resolution mass spectrum data；
Described extract is carried out mass spectral analysis under time of-flight mass spectrometer anion full scan pattern,
The second one-level high resolution mass spectrum data and the two or two grade of high resolution mass spectrum data to extract；
The first one-level high resolution mass spectrum data according to extract, the one or two grade of high resolution mass spectrum of extract
Data, the second one-level high resolution mass spectrum data of extract, the two or two grade of high resolution mass spectrum number of extract
According to step S1) in high resolution mass spectrum database detection testing sample in antioxidant and preservative.
Method the most according to claim 1, it is characterised in that described step S1) particularly as follows:
The list mark methanol solution of preparation antioxidant；
The list mark methanol solution of described antioxidant is carried out liquid-phase chromatographic analysis, obtains the liquid of antioxidant
Phase chromatographic data；
By the list mark methanol solution of described antioxidant under time of-flight mass spectrometer cation full scan pattern
Carry out mass spectral analysis, the first one-level high resolution mass spectrum data of collection antioxidant and the one or two grade of high-resolution
Mass spectrometric data；
By the list mark methanol solution of described antioxidant under time of-flight mass spectrometer anion full scan pattern
Carry out mass spectral analysis, the second one-level high resolution mass spectrum data of collection antioxidant and the two or two grade of high-resolution
Mass spectrometric data；
The list mark methanol solution of preparation preservative；
The list mark methanol solution of described preservative is carried out liquid-phase chromatographic analysis, obtains the liquid phase look of preservative
Modal data；
The list mark methanol solution of described preservative is entered under time of-flight mass spectrometer cation full scan pattern
Row mass spectral analysis, the first one-level high resolution mass spectrum data of collection preservative and the one or two grade of high resolution mass spectrum
Data；
The list mark methanol solution of described preservative is entered under time of-flight mass spectrometer anion full scan pattern
Row mass spectral analysis, the second one-level high resolution mass spectrum data of collection preservative and the two or two grade of high resolution mass spectrum
Data；
Setting up high resolution mass spectrum database, described high resolution mass spectrum database includes the 1st of antioxidant
Level high resolution mass spectrum data, the one or two grade of high resolution mass spectrum data of antioxidant, the second of antioxidant
One-level high resolution mass spectrum data, the two or two grade of high resolution mass spectrum data of antioxidant, the first of preservative
One-level high resolution mass spectrum data, the one or two grade of high resolution mass spectrum data of preservative, the 2nd 1 of preservative the
Level high resolution mass spectrum data and the two or two grade of high resolution mass spectrum data of preservative.
Method the most according to claim 1, it is characterised in that described step S2 particularly as follows:
Testing sample alcoholic solution is extracted, obtains extract；
Described extract is first carried out liquid-phase chromatographic analysis, obtains the Liquid Chromatography data of extract；
Extract through liquid-phase chromatographic analysis is carried out in time of-flight mass spectrometer cation full scan pattern
Mass spectral analysis, obtains the first one-level high resolution mass spectrum data of extract and the one or two grade of high resolution mass spectrum number
According to；
Extract through liquid-phase chromatographic analysis is entered under time of-flight mass spectrometer anion full scan pattern
Row mass spectral analysis, obtains the second one-level high resolution mass spectrum data of extract and the two or two grade of high resolution mass spectrum
Data；
Liquid Chromatography data according to extract, the first one-level high resolution mass spectrum data of extract, extraction
One or two grade of high resolution mass spectrum data of liquid, the second one-level high resolution mass spectrum data of extract, extract
The two or two grade of high resolution mass spectrum data and step S1) in high resolution mass spectrum database detection testing sample
In antioxidant and preservative.
The most according to the method in claim 2 or 3, it is characterised in that described liquid-phase chromatographic analysis
Mobile phase A is formic acid water, and Mobile phase B is the mixed liquor of methyl alcohol and formic acid, and the Mobile phase B of 70% is isocratic
Wash-out.
Method the most according to claim 1, it is characterised in that described time of-flight mass spectrometer just from
The Mass Spectrometry Conditions of sub-full scan pattern is: electron spray ESI ion gun；Capillary voltage 3000～4000V；
Atomization gas 1～3Bar；It is dried temperature 150 DEG C～200 DEG C；Dry gas stream speed 2～10l/min；Quality is swept
Retouch scope m/z 50～1000amu；Six grades of bar radio-frequency voltages 150.0～300.0Vpp；Collision cell radio-frequency voltage
500.0～800.0Vpp.
Method the most according to claim 1, it is characterised in that described time of-flight mass spectrometer bear from
The Mass Spectrometry Conditions of sub-full scan pattern is: electron spray ESI ion gun；Capillary voltage 2500～3500V；
Atomization gas 1～3Bar；It is dried temperature 150 DEG C～200 DEG C；Dry gas stream speed 2～10l/min；Quality is swept
Retouch scope m/z 50～1000amu；Level Four bar ion energy 2.0～6.0eV；Collision cell penetrates energy 5.0～10.0
eV；Collision cell radio-frequency voltage 500.0～800.0Vpp；Transmission time 80.0～120.0 μ s；Prefocus frequency
3.0～7.0 μ s.
Method the most according to claim 1, it is characterised in that described step S1) in one-level high score
Distinguish that the mass spectrometric data accurate mass number error with two grades of high resolution mass spectrum data is less than or equal to 2mDa, coordination
Element peak shape deviation is less than or equal to 20.
Preparation method the most according to claim 1, it is characterised in that described antioxidant and anticorrosion
Agent be thiabendazole, ethoxyquinoline, glycyrrhizic acid, p-Coumaric Acid, propylgallate, natamycin,
Methyl hydroxybenzoate, ethylparaben, propylparaben, n Heptyl p hydroxybenzoate, to phenyl
Phenol, benzyl p-hydroxybenzoate, 4-HBA butyl ester, 4-HBA isobutyl ester, 4-hexyl
Resorcinol, 2,4-D acid, P-hydroxybenzoic acid n-octyl, forulic acid, TBHQ, 3,3-
Dithiodipropionic acid dilauryl ester, Rosmarinic acid and ascorbic acid.
Preparation method the most according to claim 1, it is characterised in that described testing sample is smokeless
Tobacco.
10. an antioxidant or the method for building up of preservative high resolution mass spectrum database, it is characterised in that
Including:
The list mark methanol solution of preparation antioxidant；
By the list mark methanol solution of described antioxidant under time of-flight mass spectrometer cation full scan pattern
Carry out mass spectral analysis, the first one-level high resolution mass spectrum data of collection antioxidant and the one or two grade of high-resolution
Mass spectrometric data；
By the list mark methanol solution of described antioxidant under time of-flight mass spectrometer anion full scan pattern
Carry out mass spectral analysis, the second one-level high resolution mass spectrum data of collection antioxidant and the two or two grade of high-resolution
Mass spectrometric data；
The list mark methanol solution of preparation preservative；
The list mark methanol solution of described preservative is entered under time of-flight mass spectrometer cation full scan pattern
Row mass spectral analysis, the first one-level high resolution mass spectrum data of collection preservative and the one or two grade of high resolution mass spectrum
Data；
The list mark methanol solution of described preservative is entered under time of-flight mass spectrometer anion full scan pattern
Row mass spectral analysis, the second one-level high resolution mass spectrum data of collection preservative and the two or two grade of high resolution mass spectrum
Data；
Setting up high resolution mass spectrum database, described high resolution mass spectrum database includes the 1st of antioxidant
Level high resolution mass spectrum data, the one or two grade of high resolution mass spectrum data of antioxidant, the second of antioxidant
One-level high resolution mass spectrum data, the two or two grade of high resolution mass spectrum data of antioxidant, the first of preservative
One-level high resolution mass spectrum data, the one or two grade of high resolution mass spectrum data of preservative, the 2nd 1 of preservative the
Level high resolution mass spectrum data and the two or two grade of high resolution mass spectrum data of preservative.
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| CN106770803A (en) * | 2017-02-27 | 2017-05-31 | 江苏江大源生态生物科技股份有限公司 | Polyphenols HPLC/Q TOF MS detection methods in a kind of propolis |
| CN108037195A (en) * | 2017-11-20 | 2018-05-15 | 广西壮族自治区食品药品检验所 | The screening method and system of chemical composition are added in a kind of rice flour |
| CN108426953A (en) * | 2018-03-02 | 2018-08-21 | 内蒙古蒙牛乳业（集团）股份有限公司 | The method for detecting natamycin content in dairy produce |
| CN108828110B (en) * | 2018-09-11 | 2021-09-21 | 广东出入境检验检疫局检验检疫技术中心 | Method for determining residual quantity of benzimidazole drugs in chicken fat |
| CN108828110A (en) * | 2018-09-11 | 2018-11-16 | 广东出入境检验检疫局检验检疫技术中心 | The measuring method of benzimidazoles residues residual quantity in one breeder fat |
| CN110907575A (en) * | 2018-09-14 | 2020-03-24 | 中国科学院大连化学物理研究所 | Deep annotation method of hydroxycinnamic acid amide in plants |
| CN110907575B (en) * | 2018-09-14 | 2021-10-08 | 中国科学院大连化学物理研究所 | Deep annotation method of hydroxycinnamic acid amide in plants |
| CN109298063A (en) * | 2018-10-15 | 2019-02-01 | 广东工业大学 | A kind of method of quick detection facial mask natural component |
| CN111024851A (en) * | 2019-12-26 | 2020-04-17 | 江苏傲农生物科技有限公司 | Method for simultaneously detecting contents of various additives |
| CN113030324A (en) * | 2021-03-10 | 2021-06-25 | 宁波中盛产品检测有限公司 | Method for detecting preservative hydroxy ethoxy phenyl butanone in infant care product |
| CN113933405A (en) * | 2021-09-02 | 2022-01-14 | 大海粮油工业（防城港）有限公司 | Method for detecting content of tea polyphenol palmitate based on liquid chromatograph |
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