CN111650307A - Method for simultaneously qualitatively and quantitatively detecting illegal additives of antipyretic and analgesic drugs in wine and application - Google Patents
Method for simultaneously qualitatively and quantitatively detecting illegal additives of antipyretic and analgesic drugs in wine and application Download PDFInfo
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Abstract
The invention discloses a method for qualitatively and quantitatively detecting illegal additives of antipyretic and analgesic drugs in wine and application thereof. The method comprises the following steps: preparing standard series solutions of four illegal additives, namely caffeine, acetaminophen, aspirin and iprodione; heating wine, removing volatile substances, cooling, and adding acetonitrile to complement the mass to obtain a sample solution to be detected; determining each standard series solution by adopting a liquid chromatography-tandem mass spectrometry method, and drawing a standard curve to obtain a regression equation A which is kC + b; and detecting the sample solution to be detected, and substituting the peak area A of the sample into a regression equation to obtain the concentration of each illegal additive in the sample. The method has the advantages of simple and convenient pretreatment, strong specificity and high sensitivity, can simultaneously carry out qualitative and quantitative analysis on the caffeine, the aspirin, the acetaminophen and the isopropanyline, and can be applied to food safety supervision to prevent the illegal addition of the four medicines in the wine.
Description
Technical Field
The invention relates to the technical field of analytical chemistry, in particular to a method for qualitatively and quantitatively detecting illegal additives of antipyretic and analgesic drugs in wine and application thereof, and specifically relates to a method for qualitatively and quantitatively analyzing caffeine, aspirin, acetaminophen and ipratropium in wine and application thereof.
Background
The fermented wine is fermented by grains or fruits, and the prepared wine is prepared by infusing the fermented wine and medicinal materials or fruits, so that the wine industry is an extension of the agricultural product industry and is also an important supporting industry of the agricultural product industry. While the headache of the drinker is one of the most headache caused by the intoxication or the alcohol drinking. In order to meet the concept that grain wine cannot be sold in the market, illegal merchants add antipyretic and analgesic drugs into white spirit, and the impact force on the white spirit industry is large and the negative effect is serious. The most common antipyretic and analgesic drugs on the market comprise acanthophenol powder and sanli pain, and the like, wherein the main components of the acanthophenol powder comprise caffeine, acetaminophen and aspirin (namely acetylsalicylic acid), and the main components of the sanli pain comprise caffeine, acetaminophen and ipratropium. The two medicines can stimulate gastric mucosa when taken together with wine, and may cause liver injury when taken in large quantities. Therefore, the wine cannot enter the dining table of people.
In order to prevent the liquor from being circulated to the market, the supervision of the commodities should be strengthened, but at present, a method for qualitatively and quantitatively analyzing the several substances is not available.
The qualitative and quantitative methods mainly comprise thin layer chromatography, ultraviolet method, liquid chromatography-diode detector combined method, liquid chromatography-mass spectrometry and the like. The thin-layer chromatography has low accuracy, is greatly influenced by matrix, has low precision, and can not be basically detected by samples lower than 1 g/L; the accuracy of the ultraviolet method is not high, the influence of the matrix is large, and the method can not be used for determining the quality basically when a plurality of components are mixed; the liquid chromatogram-diode detector combined method has low precision, and samples with the concentration lower than 1mg/L can not be basically detected; liquid chromatography-mass spectrometry is widely applied internationally, but the application of mass spectrometry for qualitative and quantitative analysis is limited to caffeine and acetaminophen, while isoalanobine and acetylsalicylic acid have no mass spectrometry quantitative analysis method.
Therefore, in order to improve the detection efficiency and realize the simultaneous qualitative and quantitative analysis, the invention develops a method for simultaneously carrying out qualitative and quantitative analysis on the four components of caffeine, acetaminophen, iprodione and acetylsalicylic acid by using a liquid chromatography-mass spectrometry combined technology.
Disclosure of Invention
In view of the defects of the prior art, the invention aims to provide a method for qualitatively and quantitatively detecting illegal additives of antipyretic and analgesic drugs in wine and application thereof, researches a method for qualitatively and quantitatively analyzing four components simultaneously by using a liquid chromatography-mass spectrometry combined technology, and fills the gap of simultaneously qualitatively and quantitatively detecting trace caffeine, acetaminophen, aspirin and isopropaneantipyrine in wine at home and abroad.
In order to achieve the above objects and other related objects, the present invention provides a method for simultaneously qualitatively and quantitatively detecting illegal additives of antipyretic and analgesic drugs in wine, wherein the illegal additives are caffeine, acetaminophen, aspirin and ipratropium, the method comprising the steps of:
(1) preparation of a standard series of solutions: respectively preparing standard stock solutions of four illegal additives, diluting the standard stock solutions into standard use solutions, and preparing the standard use solutions into standard series solutions by using acetonitrile aqueous solutions;
(2) sample pretreatment: heating wine, removing volatile substances, cooling, and adding acetonitrile to complement the mass to obtain a sample solution to be detected;
(3) drawing a standard curve: measuring standard series solutions of the four illegal additives by adopting a liquid chromatography-tandem mass spectrometry method, and drawing standard curves of the four illegal additives according to the peak area A of a standard point and the concentration C of the corresponding standard series solution to obtain a regression equation A which is kC + b;
(4) and (3) sample determination: and (4) detecting the solution of the sample to be detected by adopting a liquid chromatography-tandem mass spectrometry method to obtain the peak area A of the sample to be detected, substituting the peak area A into the standard curve regression equation in the step (3), and calculating to obtain the concentrations of the four illegal additives in the sample to be detected.
Further, in the step (1), the concentrations of the standard stock solutions of caffeine, aspirin, acetaminophen and isopropylantipyrine are 0.5 + -0.05 mg/ml, 1 + -0.1 mg/ml, 0.1 + -0.01 mg/ml and 1 + -0.1 mg/ml, respectively.
Further, in the step (1), the standard stock solution is diluted by 100-fold and 1000-fold to be used as the standard using solution.
Further, in the step (1), the solvent used for preparing the standard stock solution and the standard use solution is water or methanol.
Optionally, in the step (1), the water is ultrapure water, the methanol is chromatographically pure methanol, and the acetonitrile aqueous solution is 60% acetonitrile aqueous solution.
Further, in the step (1), the concentration range of the standard series solution is 10-1000ng/ml, and no less than five points of the standard series solution are prepared for each illegal additive.
Optionally, in the step (1), the concentrations of the standard series of solutions of caffeine are, in order: 20. 50, 80, 100 and 200ng/ml of standard series solutions of acetaminophen, wherein the concentrations of the standard series solutions of acetaminophen are as follows: 10. 20, 50, 80, 100, 200ng/ml, the concentrations of the standard series of solutions of isopropylamine tipelin are, in order: 40. 60, 80, 100 and 200ng/ml, and the concentration of the standard series solutions of aspirin is as follows in sequence: 400. 500, 600, 800 and 1000 ng/ml.
Further, in the step (2), the wine is distilled in a boiling water bath to remove volatile substances. The main volatile substance in the wine is ethanol.
Further, in the step (2), the sample solution to be detected is filtered by a filter membrane and then detected, wherein the filter membrane is a 0.25 μm microporous filter membrane.
Further, in the steps (3) and (4), the liquid chromatography determination conditions are as follows:
mobile phase: acetonitrile and water, and gradient elution; the liquid chromatographic column is ZORBAX Eclipsee plus C18 Rapidresolution HD, 2.1X 50mm,1.8 μm or equivalent liquid chromatographic column; column temperature: 20-35 ℃; flow rate: 0.3 ml/min; sample introduction amount: 10 μ L.
Further, the mobile phase comprises 10-90-% acetonitrile and 90-10% water, the water contains an ionization aid, the ionization aid comprises formic acid and ammonium formate, the formic acid is 0.1% formic acid, and the ammonium formate is 0.01mmol/L ammonium formate.
Further, in the liquid chromatography determination conditions, the separation degree is more than 1.5, and the gradient elution procedure of the mobile phase is as follows:
0-3min, wherein the volume ratio of acetonitrile to water is 1:9-1: 9;
3-5.1min, wherein the volume ratio of acetonitrile to water is 1:9-8: 2;
5.1-8min, wherein the volume ratio of acetonitrile to water is 8:2-9: 1;
8-12min, wherein the volume ratio of acetonitrile to water is 9:1-9: 1.
Further, in the steps (3) and (4), the mass spectrometry conditions are as follows:
the mass spectrometer is Agilent 6460QQQ and/or a detector with equivalent function; an ion source: ESI, cone voltage: 4000 KV; flow rate of drying gas: 11l/min, assist gas pressure: 15 psi; temperature of the drying gas: at 150 ℃.
Further, under the mass spectrometry conditions, the peak-off time, the quantitative ion pair, the shaped ion pair, the fragmentation voltage, the collision voltage, and the detection mode of each illegal additive are respectively as follows:
acetaminophen: 1.2min, 152-110, 114V, 15V, positive ion;
isopropyl antipyrine: 5.9min, 231, 189, 92, 17V, positive ion;
and/or, iprodione: 5.9min, 231, 112, 21, 15V, positive ion;
caffeine: 2.0min, 195-138, 100V, 20V, positive ion;
and/or, caffeine: 2.0min, 195-138, 195-110, 100V, 20V, positive ion;
aspirin: 4.9min, 179 and 137, 45V, 5V, negative ions.
On the other hand, the invention provides the application of the method for qualitatively and quantitatively detecting the illegal additives of the antipyretic and analgesic medicines in the liquor in detection of the illegal additives of the antipyretic and analgesic medicines in the liquor.
As mentioned above, the method for qualitatively and quantitatively detecting illegal additives of antipyretic and analgesic drugs in wine and the application thereof have the following beneficial effects:
the invention discloses a liquid chromatography-tandem mass spectrometry analysis method for simultaneously carrying out qualitative and quantitative detection on four illegal additives, namely caffeine, aspirin, acetaminophen and isopropanamiline in white spirit. The method uses an ultra-high performance liquid chromatograph as separation equipment, a triple quadrupole mass spectrometer as a detector, and performs qualitative analysis by using mass spectrum qualitative ion pairs, and performs quantitative detection by using quantitative ions for an external standard method. The method has the advantages of simple and convenient pretreatment, strong specificity and high sensitivity, can simultaneously carry out qualitative and quantitative analysis on the four substances of caffeine, aspirin, acetaminophen and isopropylphenazone, and can be applied to food safety supervision to prevent the illegal addition of the four antipyretic analgesic drugs in wine.
Drawings
FIG. 1 is a standard graph of caffeine in an embodiment of the present invention.
FIG. 2 is a graph showing a standard curve of acetylsalicylic acid in the examples of the present invention.
FIG. 3 is a graph showing a standard curve of acetaminophen in the examples of the present invention.
FIG. 4 is a graph showing a standard curve of ipratropium in the example of the present invention.
FIG. 5 shows the extracted ion chromatograms of standard series solutions of different concentrations of caffeine in examples of the present invention.
FIG. 6 shows the extracted ion chromatograms of standard series solutions of acetylsalicylic acid of different concentrations in the examples of the present invention.
FIG. 7 shows the extracted ion chromatograms of standard series solutions of acetaminophen at different concentrations in the examples of the present invention.
FIG. 8 shows the extracted ion chromatograms of different concentrations of the standard series solutions of isopropylantipyrine in the examples of the present invention.
FIG. 9 shows the mass spectrum of caffeine in an example of the present invention.
FIG. 10 shows the mass spectrum of acetylsalicylic acid in the present example.
FIG. 11 shows a mass spectrum of acetaminophen in the examples of the present invention.
FIG. 12 shows the mass spectrum of ipratropium in the present example.
FIG. 13 shows a total ion current chromatogram of a blank sample in an example of the present invention.
FIG. 14 shows a total ion current chromatogram of a negative sample in an example of the present invention.
FIG. 15 shows a total ion current chromatogram of a negative spiked sample in an example of the invention.
FIG. 16 shows a total ion current chromatogram of a standard sample in an example of the present invention.
Detailed Description
The embodiments of the present invention are described below with reference to specific embodiments, and other advantages and effects of the present invention will be easily understood by those skilled in the art from the disclosure of the present specification. The invention is capable of other and different embodiments and of being practiced or of being carried out in various ways, and its several details are capable of modification in various respects, all without departing from the spirit and scope of the present invention.
The invention provides a method for qualitatively and quantitatively detecting illegal additives of antipyretic and analgesic drugs in wine, wherein the illegal additives are caffeine, acetaminophen, aspirin and isopropanoantipyrine, and the method comprises the following specific steps:
1. preparing standard series solution
(1) Standard stock solutions of caffeine, aspirin, acetaminophen and iprantelopine were prepared with specific dilution solvents, preparation volumes and concentrations as given in table 1 below:
TABLE 1 stock solution compounding table
(2) The standard stock solution in step (1) was diluted 1000-fold with methanol to serve as a standard use solution.
(3) Preparing standard series solutions with the concentration of 10-1000ng/L from the standard use solution in the step (2) by using a 60% acetonitrile water solution, wherein the standard points are not less than 5, and specifically are respectively as follows:
caffeine: 20. 50, 80, 100, 200ng/ml,
acetaminophen: 10. 20, 50, 80, 100, 200ng/ml,
isopropylamine tipelin: 40. 60, 80, 100, 200ng/ml,
aspirin: 400. 500, 600, 800 and 1000 ng/ml.
(4) The standard series of solutions was filtered through a 0.25 μm filter and used.
2. Sample pretreatment
Distilling the sample to be tested in boiling water bath for 20min, removing volatile substances (ethanol), cooling, adding acetonitrile to complement the mass, and filtering with 0.25 μm microporous membrane to be tested.
3. Liquid chromatography-tandem mass spectrometry assay
(1) Conditions for liquid chromatography
Mobile phase: acetonitrile and water (0.01mmol/L ammonium formate, 0.1% formic acid), gradient elution, the order of elution being given in table 2 below; column temperature: 20-35 ℃; flow rate: 0.3 ml/min; sample introduction amount: 10 μ L.
TABLE 2 gradient elution sequence
| Time (min) | Acetonitrile | Water (0.01mmol/L ammonium formate, 0.1% formic acid) |
| 0 | 10％ | 90％ |
| 3 | 10％ | 90％ |
| 5.1 | 80％ | 20％ |
| 8 | 90％ | 10％ |
| 12 | 90％ | 10％ |
(2) Conditions for Mass Spectrometry
The mass spectrometer model: agilent 6460 qqqq or equivalent detector; an ion source: ESI, cone voltage: 4000KV, dry air flow: 11l/min, assist gas pressure: 15 psi; temperature of the drying gas: 150 ℃; the qualitative and quantitative ion pairs and the detection mode are shown in table 3 below.
TABLE 3 Mass Spectrometry detection conditions Table
(3) Drawing a standard curve: and respectively injecting the standard series solutions of the illegal additives into a liquid chromatogram-tandem mass spectrometer, and drawing a standard curve according to the peak area (A) of the standard point and the corresponding concentration (C) of the standard point to obtain a regression equation A which is kC + b, wherein b is a vertical coordinate.
The standard curves for caffeine, aspirin, acetaminophen and ipratropium are shown in fig. 1, fig. 2, fig. 3 and fig. 4, respectively.
The standard curve for caffeine is: a. theCaffeine＝15335C+12.227，R2＝0.9995；
The standard curve for acetylsalicylic acid is: a. theAcetylsalicylic acid＝32.865C-30.175，R20.9985; (acetylsalicylic acid is known as aspirin in the name of medical drug)
The standard curve for acetaminophen is: a. theAcetaminophen＝9.4806C+421.25，R2＝0.9974；
The standard curve for ipratropium is: a. theIsopropinebrine＝8.8559C+57.596，R2＝0.9985。
The extracted ion chromatograms of caffeine, aspirin, acetaminophen, and ipratropium are shown in fig. 5, 6, 7, and 8, respectively.
The mass spectra of caffeine, aspirin, acetaminophen and ipratropium are shown in fig. 9, fig. 10, fig. 11 and fig. 12, respectively.
(4) And detecting the solution of the sample to be detected by adopting a liquid chromatography-tandem mass spectrometry method to obtain the peak area A of the sample to be detected, and substituting the peak area A of the sample to be detected into a standard curve regression equation to obtain the concentrations of the four illegal additives in the sample to be detected.
Example 1
The method is adopted to carry out addition and recovery experiments on acetaminophen, aspirin, caffeine and isopropyl antipyrine in 12 types of white spirits respectively, wherein the 12 types of white spirits respectively have strong fragrance type, faint scent type, maotai-flavor type, phoenix flavor type, fermented soybean flavor type, compatible flavor type, medicinal flavor type, special flavor type, rice flavor type, sesame flavor type, strong fragrance flavor type and white spirit, and the white spirits for various types of experiments and manufacturers are shown in Table 13:
TABLE 13 white spirit statistics table for experiment
Example 2
Recovery rate experiment
Accurately weighing 20g of 12 negative samples, sucking a certain volume of standard solution, adding the standard solution into the samples, processing according to a sample pretreatment method, measuring, then calculating the recovery rate of the added samples, and expressing the precision of the method by relative standard deviation. The concentrations (normalized concentrations) and normalized volumes of the standard solutions added to the samples are shown in tables 4 to 7, and the recovery rates and their relative standard deviations are shown in tables 8 to 11.
Table 4 caffeine standard solution concentrations: 10 ug/ml
| Standard concentration of | 80ng/g | 120ng/g | 200ng/g |
| Adding the standard liquid | 160μl | 240μl | 400μl |
Table 5 aspirin (acetylsalicylic acid) standard solution concentrations: 800 ug/ml
| Standard concentration of | 4μg/g | 8μg/g | 10μg/g |
| Adding the standard liquid | 100μl | 200μl | 250μl |
Table 6 acetaminophen standard solution concentrations: 10 ug/ml
| Standard concentration of | 50ng/g | 80ng/g | 100ng/g |
| Adding the standard liquid | 100μl | 160μl | 200μl |
Table 7 concentration of the standard solution of isopropylantipyrine: 10 ug/ml
| Standard concentration of | 80ng/g | 120ng/g | 200ng/g |
| Adding the standard liquid | 160μl | 240μl | 400μl |
TABLE 8 recovery of caffeine by spiking
| Standard concentration of | 80ng/g | 120ng/g | 200ng/g |
| Recovery rate | 92.1～110.7％ | 95.3～107.0％ | 96.7～105.1％ |
| Relative Standard Deviation (SD) | 85.7～107.5％ | 86.8～105.8％ | 89.6～104.3％ |
TABLE 9 recovery of aspirin (acetylsalicylic acid) on the normalized scale
| Standard concentration of | 4μg/g | 8μg/g | 10μg/g |
| Recovery rate | 83.8～116.7％ | 89.6～109.0％ | 90.7～107.3％ |
| Relative Standard Deviation (SD) | 95.7～102.5％ | 96.2～104.1％ | 95.6～103.1％ |
TABLE 10 recovery of acetaminophen on a standard basis
| Standard concentration of | 50ng/g | 80ng/g | 100ng/g |
| Recovery rate | 83.5～114.7％ | 88.4～111.2％ | 90.7～105.3％ |
| Relative Standard Deviation (SD) | 83.2～109.1％ | 85.7～107.5％ | 88.3～105.6％ |
TABLE 11 recovery of iprodione with spiking
| Standard concentration of | 80ng/g | 120ng/g | 200ng/g |
| Recovery rate | 94.1～109.1％ | 95.3～106.8％ | 96.2～105.4％ |
| Relative Standard Deviation (SD) | 91.7～104.7％ | 92.8～103.6％ | 93.6～102.8％ |
Example 3
Minimum detection limit test
The standard solution was diluted with 60% ethanol aqueous solution and measured, and when the peak height of the component to be measured was 3 times the signal-to-noise ratio, the corresponding concentration was the lowest detection limit of the component, and the results are shown in table 12.
TABLE 12 minimum detection limits for the components to be tested
| Component to be tested | Caffeine | Acetylsalicylic acid | Acetaminophen | Isopropinebrine |
| Minimum limit of detection | 50ng/ml | 1μg/ml | 20ng/ml | 40ng/ml |
The experimental results show that the total ion current chromatogram (shown in figure 14) of the negative sample of the 12 kinds of fragrance liquor treated by the method has no obvious impurity peak compared with the total ion current chromatogram (shown in figure 13) of the blank sample; and the total ion current chromatogram of the negative labeled sample (as shown in fig. 15) has a significant peak increase compared with the total ion current chromatogram of the blank sample (as shown in fig. 13), and the retention time and the mass spectrum of the increased peak are consistent with those of the target peak in the total ion current chromatogram of the standard sample (as shown in fig. 16). Moreover, as can be seen from the recovery rate test of example 2, the method has good recovery rate and relative standard deviation (see tables 8-11), which shows that the method of the invention can be used for quickly and effectively carrying out qualitative and quantitative determination on acetaminophen, aspirin, caffeine and isopropanetipyrin in white spirit.
In conclusion, the invention provides a liquid chromatography-tandem mass spectrometry analysis method capable of simultaneously carrying out qualitative and quantitative detection on four illegal additives, namely caffeine, aspirin, acetaminophen and isopropanylidene in white spirit. The method uses an ultra-high performance liquid chromatograph as separation equipment, a triple quadrupole mass spectrometer as a detector, and performs qualitative analysis by using mass spectrum qualitative ion pairs, and performs quantitative detection by using quantitative ions for an external standard method. The method has the advantages of simple and convenient pretreatment, strong specificity and high sensitivity, can simultaneously carry out qualitative and quantitative analysis on the four substances of caffeine, aspirin, acetaminophen and isopropylphenazone, and can be applied to food safety supervision to prevent the illegal addition of the four antipyretic analgesic drugs in wine.
The foregoing embodiments are merely illustrative of the principles and utilities of the present invention and are not intended to limit the invention. Any person skilled in the art can modify or change the above-mentioned embodiments without departing from the spirit and scope of the present invention. Accordingly, it is intended that all equivalent modifications or changes which can be made by those skilled in the art without departing from the spirit and technical spirit of the present invention be covered by the claims of the present invention.
Claims (10)
1. A method for qualitatively and quantitatively detecting illegal additives of antipyretic and analgesic drugs in wine simultaneously is characterized in that the illegal additives are caffeine, acetaminophen, aspirin and isopropanoantipyrine, and the method comprises the following steps:
(1) preparation of a standard series of solutions: respectively preparing standard stock solutions of four illegal additives, diluting the standard stock solutions into standard use solutions, and preparing the standard use solutions into standard series solutions by using acetonitrile aqueous solutions;
(2) sample pretreatment: heating wine, removing volatile substances, cooling, and adding acetonitrile to complement the mass to obtain a sample solution to be detected;
(3) drawing a standard curve: measuring standard series solutions of the four illegal additives by adopting a liquid chromatography-tandem mass spectrometry method, and drawing standard curves of the four illegal additives according to the peak area A of a standard point and the concentration C of the corresponding standard series solution to obtain a regression equation A which is kC + b;
(4) and (3) sample determination: and (4) detecting the solution of the sample to be detected by adopting a liquid chromatography-tandem mass spectrometry method to obtain the peak area A of the sample to be detected, substituting the peak area A into the standard curve regression equation in the step (3), and calculating to obtain the concentrations of the four illegal additives in the sample to be detected.
2. The method of claim 1, wherein: in the step (1), the concentrations of the standard stock solutions of the caffeine, the aspirin, the acetaminophen and the isopropantipyrine are respectively 0.5 +/-0.05 mg/ml, 1 +/-0.1 mg/ml, 0.1 +/-0.01 mg/ml and 1 +/-0.1 mg/ml;
and/or, in the step (1), the standard stock solution is diluted by 100-fold and 1000-fold to be used as the standard using solution;
and/or, in the step (1), the solvent used for preparing the standard stock solution and the standard use solution is water or methanol;
and/or, in the step (1), the concentration range of the standard series solution is 10-1000ng/ml, and no less than five points of the standard series solution are prepared for each illegal additive.
3. The method of claim 2, wherein: in the step (1), water is ultrapure water, methanol is chromatographically pure methanol, and the acetonitrile water solution is 60% acetonitrile water solution.
4. The method of claim 1, wherein: in the step (2), distilling the wine in a boiling water bath to remove volatile substances;
and/or, in the step (2), the sample solution to be detected is filtered by a filter membrane and then detected, wherein the filter membrane is a 0.25 mu m microporous filter membrane.
5. The method of claim 1, wherein: in the steps (3) and (4), the liquid chromatography determination conditions are as follows:
mobile phase: acetonitrile and water, and gradient elution; the liquid chromatographic column is ZORBAX Eclipsee plus C18 Rapidresolution HD, 2.1X 50mm,1.8 μm or equivalent liquid chromatographic column; column temperature: 20-35 ℃; flow rate: 0.3 ml/min; sample introduction amount: 10 μ L.
6. The method of claim 4, wherein: the mobile phase comprises 10-90-% acetonitrile and 90-10% water, the water contains an ionization auxiliary agent, the ionization auxiliary agent comprises formic acid and ammonium formate, the formic acid is 0.1% formic acid, and the ammonium formate is 0.01mmol/L ammonium formate.
7. The method of claim 4, wherein: in the liquid chromatogram determination conditions, the separation degree is more than 1.5, and the gradient elution procedure of the mobile phase is as follows:
0-3min, wherein the volume ratio of acetonitrile to water is 1:9-1: 9;
3-5.1min, wherein the volume ratio of acetonitrile to water is 1:9-8: 2;
5.1-8min, wherein the volume ratio of acetonitrile to water is 8:2-9: 1;
8-12min, wherein the volume ratio of acetonitrile to water is 9:1-9: 1.
8. The method of claim 1, wherein: in the steps (3) and (4), the mass spectrometry conditions are as follows: the mass spectrometer is Agilent 6460QQQ and/or a detector with equivalent function; an ion source: ESI, cone voltage: 4000 KV; flow rate of drying gas: 11l/min, assist gas pressure: 15 psi; temperature of the drying gas: at 150 ℃.
9. The method of claim 8, wherein: in the mass spectrometry conditions, the peak-off time, the quantitative ion pair, the shaped ion pair, the fragmentation voltage, the collision voltage and the detection mode of each illegal additive are respectively as follows:
acetaminophen: 1.2min, 152-110, 114V, 15V, positive ion;
isopropyl antipyrine: 5.9min, 231, 189, 92, 17V, positive ion;
and/or, iprodione: 5.9min, 231, 112, 21, 15V, positive ion;
caffeine: 2.0min, 195-138, 100V, 20V, positive ion;
and/or, caffeine: 2.0min, 195-138, 195-110, 100V, 20V, positive ion;
aspirin: 4.9min, 179 and 137, 45V, 5V, negative ions.
10. The use of the method according to any one of claims 1 to 9 for simultaneously qualitatively and quantitatively detecting the illegal addition of antipyretic and analgesic drugs in wine for detecting the illegal addition of antipyretic and analgesic drugs in white spirit.
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