CN113295797A - Method for rapidly detecting ethyl carbamate in white spirit based on ultra-high performance liquid chromatography combined high-resolution mass spectrometry - Google Patents
Method for rapidly detecting ethyl carbamate in white spirit based on ultra-high performance liquid chromatography combined high-resolution mass spectrometry Download PDFInfo
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- G01N30/14—Preparation by elimination of some components
- G01N2030/146—Preparation by elimination of some components using membranes
Abstract
The invention discloses a method for rapidly detecting ethyl carbamate in white spirit based on ultra-high performance liquid chromatography combined high resolution mass spectrometry, and belongs to the technical field of detection and screening of dangerous substances in fermented food. The method comprises the following steps: carrying out concentration and redissolution pretreatment on the white spirit sample, and extracting and enriching a target substance to be detected in the white spirit sample to obtain a target substance to be detected sample; detecting the obtained target sample to be detected based on the ultra-high performance liquid chromatography and high resolution mass spectrometry technology to obtain a primary mass spectrogram and a secondary mass spectrogram of the target sample to be detected; extracting target object information through software based on the obtained primary and secondary mass spectrograms: and establishing a standard spectrogram based on the ethyl carbamate standard substance, and comparing the established standard spectrogram with the obtained extracted target object information to qualitatively and quantitatively determine the ethyl carbamate in the white spirit sample. A systematic detection method of food safety risk substance ethyl carbamate in white spirit with high sensitivity and high accuracy is established.
Description
Technical Field
The invention belongs to the technical field of detection and screening of dangerous substances in fermented foods, and particularly relates to a method for rapidly detecting ethyl carbamate in white spirit based on ultra-high performance liquid chromatography combined high-resolution mass spectrometry.
Background
The white spirit is the traditional distilled spirit in China, is one of the six distilled spirits in the world, is unique in the distilled spirits in the world by the unique process and style, and is deeply loved by the masses of drinkers. At present, China pays more and more attention to food safety, and the requirement on the safety aspect of white spirit food is continuously improved. Urethane (EC) is a potentially carcinogenic compound that is widely found in traditional fermented foods. In the process of brewing white spirit, nitrogen-containing compounds in part of raw and auxiliary materials can generate ethyl carbamate in the processes of fermentation, distillation, aging and the like, and become potential food safety risks in the white spirit. The detection method of the ethyl carbamate commonly used at present mainly comprises a gas chromatography-mass spectrometry (GC-MS) and a high performance liquid chromatography fluorescence detection method (HPLC-FLD). GC-MS is a conventional technology for detecting EC in fermented food, but the method has complicated sample pretreatment, and the appropriateness of sample preparation is directly related to the result of detection analysis, so that the application range of the technology is limited to a certain extent. Compared with GC-MS, the method for analyzing the ethyl carbamate by the pre-column derivatization HPLC-FID method has higher sensitivity, accuracy and reproducibility, but the defects of complicated sample pretreatment and insufficient stability of the derivatization reagent also limit the development of the technology. The high performance liquid chromatography and high resolution mass spectrometry combined technology combines the effective separation thermal instability and high boiling point compound separation capability of a liquid chromatograph and the strong component identification capability of a mass spectrometer, is an effective means for separating and analyzing complex organic mixtures, can greatly shorten the sample pretreatment time, and meets the accurate and rapid detection requirements of wine enterprises and food safety departments on ethyl carbamate in white spirit.
Disclosure of Invention
In order to overcome the defects of the prior art, the invention aims to provide a method for rapidly detecting ethyl carbamate in white spirit based on ultra-high performance liquid chromatography combined high-resolution mass spectrometry, and a method for detecting ethyl carbamate in white spirit with high sensitivity and high accuracy by using a system for detecting food safety risk substances is established.
In order to achieve the purpose, the invention adopts the following technical scheme to realize the purpose:
the invention discloses a method for rapidly detecting ethyl carbamate in white spirit based on ultra-high performance liquid chromatography combined high resolution mass spectrometry, which comprises the following steps:
1) carrying out concentration and redissolution pretreatment on the white spirit sample, and extracting and enriching a target substance to be detected in the white spirit sample to obtain a target substance to be detected sample;
2) detecting the obtained target sample to be detected based on the ultra-high performance liquid chromatography and high resolution mass spectrometry technology to obtain a primary mass spectrogram and a secondary mass spectrogram of the target sample to be detected;
3) extracting target object information through software based on the obtained primary and secondary mass spectrograms:
4) and establishing a standard spectrogram based on the ethyl carbamate standard substance, and comparing the established standard spectrogram with the obtained extracted target object information to qualitatively and quantitatively determine the ethyl carbamate in the white spirit sample.
Preferably, the pretreatment for concentration and redissolution in step 1) specifically includes: and (3) putting 20-50mL of a white spirit sample into an evaporation bottle, rotating to evaporate to dryness, redissolving by ultrapure water, fixing the volume to 1mL, and finally filtering by using an organic phase microporous filter membrane to obtain a target sample to be detected.
Further preferably, the temperature of the rotary evaporation is 70-90 ℃.
Preferably, in the step 2), in detecting the obtained target sample to be detected based on the ultra high performance liquid chromatography combined high resolution mass spectrometry technology, specific monitoring conditions of the ultra high performance liquid chromatograph include: chromatographic column conditions: hypersil GOLD 100X 2.1mm1.9 μm; the mobile phase A is a mixed solution of water, formic acid and ammonium formate; wherein, the volume of the formic acid accounts for 0.1 percent of the volume of the mixed solution, the concentration of the ammonium formate is 4.0mmol/L, and the rest is water; the mobile phase B is a mixed solution of methanol, formic acid and ammonium formate; wherein, the volume of the formic acid accounts for 0.1 percent of the volume of the mixed solution, the concentration of the ammonium formate is 4.0mmol/L, and the rest is methanol;
the mobile phase gradient elution procedure was: within 0-1min, the volume ratio of the mobile phase A to the mobile phase B is 80%: 20% make up the mobile phase; within 1-7min, the mobile phase B is a mobile phase; within 7-12min, the mobile phase B is a mobile phase; within 12.1-13min, the volume ratio of the mobile phase A to the mobile phase B is 80%: 20% make up the mobile phase; within 13-15min, the volume ratio of the mobile phase A to the mobile phase B is 80%: 20% make up the mobile phase;
column oven: 30-40 ℃; sample introduction amount: 5-10 μ L; flow rate: 0.1-0.3 mL/min.
Preferably, in the step 2), in detecting the obtained target sample to be detected based on the ultra-high performance liquid chromatography combined high resolution mass spectrometry technology, the specific monitoring conditions of the high resolution mass spectrometer include: a detector: an electrostatic field orbital ion trap mass spectrometer; the scanning mode is as follows: ESI positive ion mode; scanning mode: full MS ddMS2(ii) a Scanning range m/z: 100-1500; spraying voltage: 3.5-5 kV; sheath gas pressure: 30-40 psi; auxiliary N2Pressure: 10-15 bar; auxiliary gas heating temperature: 300 ℃ and 400 ℃; capillary temperature: 300 ℃ and 350 ℃; lens voltage: 50-60V; full scan resolution: 70000 80000 FHWM; secondary scanning resolution: 17000 and 18000 FHWM; maximum residence time: primary full scan 150-: 50-100 ms; the normalized collision energy is: 17.5-52.5 eV.
Preferably, the specific operation of obtaining the primary mass spectrum and the secondary mass spectrum of the target sample to be tested in step 2) is as follows: and a primary mass spectrogram is obtained through a full scanning mode of a high performance liquid chromatography-ion trap orbit mass spectrum system, and a secondary mass spectrogram is obtained through scanning by selecting an ion detection mode.
Preferably, the specific operation of step 4) is: obtaining fragmentation fragment data by analyzing the secondary mass spectrogram, comparing the fragmentation fragment data with a secondary spectrogram of an ethyl carbamate standard substance, and qualitatively judging the ethyl carbamate in the white spirit; continuously analyzing other fragment information of the retention time chromatographic peak to deduce the structure and element composition of the substance; and finally, quantifying the ethyl carbamate in the white spirit sample according to the primary spectrogram of the white spirit sample and the primary spectrogram of the ethyl carbamate standard substance and the ratio of the peak area to the concentration of the extracted ion-flow diagram.
Further preferably, the specific operation of analyzing the other segment information of the retention time chromatographic peak includes: mass-to-charge ratio, fragment abundance ratio, and isotopic abundance ratio.
Further preferably, the specific operation of the ethyl carbamate standard preparation in the step 4) is as follows:
preparing a stock solution of the ethyl carbamate with the concentration of 5.00mg/mL by using a standard substance of the ethyl carbamate; preparing a urethane intermediate solution a with the concentration of 50 mug/mL by using the obtained urethane stock solution; preparing a urethane intermediate solution b with the concentration of 2.5 mu g/mL by using the obtained urethane intermediate solution a; respectively sucking the urethane intermediate liquid b: 4. mu.L, 10. mu.L, 20. mu.L, 40. mu.L, 80. mu.L and urethane intermediate liquid a: mu.L and 20. mu.L were put in 7 volumetric flasks and the volume was adjusted to 1mL to obtain 10. mu.g/L, 25. mu.g/L, 50. mu.g/L, 100. mu.g/L, 20. mu.g/L and 400. mu.g/L curve working solutions.
Wherein, the ethyl carbamate stock solution, the ethyl carbamate intermediate solution a and the ethyl carbamate intermediate solution b are further preferably stored at the temperature of below 4 ℃ for standby; the curve working solution needs to be prepared at present.
Compared with the prior art, the invention has the following beneficial effects:
the invention discloses a method for rapidly detecting ethyl carbamate in white spirit based on ultra-high performance liquid chromatography combined high-resolution mass spectrometry, which is a detection method for a system for detecting ethyl carbamate, which is a food safety risk substance in white spirit, with high sensitivity and high accuracy by taking rotary evaporation as a pretreatment means and analyzing by using ultra-high performance liquid chromatography-high resolution orbitron ion trap mass spectrometry. The method for rapidly detecting ethyl carbamate in white spirit based on ultra-high performance liquid chromatography combined high resolution mass spectrometry has the following advantages: firstly, only rotary evaporation is adopted as a pretreatment means, and a systematic screening method for establishing food safety risk substance ethyl carbamate in white spirit by using an ultra-high performance liquid chromatography-electrostatic field orbital ion trap mass spectrometry technology has the characteristics of simple pretreatment, rapidness, high efficiency, high sensitivity, low reagent cost and the like, can be used for screening and detecting ethyl carbamate in white spirit, and provides technical reference for quality safety monitoring of fermented foods. Secondly, compared with conventional detection methods such as a liquid chromatography ultraviolet method and a liquid chromatography fluorescence method, the high performance liquid chromatography and high resolution mass spectrometry combined technology has strong matrix interference resistance and more accurate substance qualitative. In addition, the method has the characteristics of low cost, high sensitivity and high accuracy, and is suitable for large-scale batch detection of samples.
Furthermore, according to the physicochemical properties of high boiling point, difficult volatilization and strong polarity of the ethyl carbamate, the efficient enrichment effect of the ethyl carbamate in the white spirit sample can be achieved in a short time by using the rotary evaporation and matching with the redissolution of ultrapure water as a unique pretreatment means, meanwhile, the interference of the matrix effect of the white spirit sample can be reduced to a certain extent, and the accuracy of the detection result is improved.
Furthermore, by using ultra-high performance liquid chromatography with a reversed phase C18 chromatographic column as a stationary phase and combining quadrupole rod-electrostatic field orbital ion trap mass spectrometry, the purposes of effectively separating thermally unstable compounds and high-boiling-point compounds and accurately identifying components can be simultaneously realized, and the method is an effective means for separating and analyzing complex organic mixtures and is suitable for accurately identifying ethyl carbamate under the background of white spirit matrix.
Drawings
FIG. 1 is a chromatogram of ethyl carbamate in a white spirit sample;
FIG. 2 is a chromatogram of a urethane standard;
FIG. 3 is a second-order mass spectrum of ethyl carbamate in a white spirit sample;
FIG. 4 is a secondary spectrum of a urethane standard.
Detailed Description
In order to make the technical solutions of the present invention better understood, the technical solutions in the embodiments of the present invention will be clearly and completely described below with reference to the drawings in the embodiments of the present invention, and it is obvious that the described embodiments are only a part of the embodiments of the present invention, and not all of the embodiments. All other embodiments, which can be derived by a person skilled in the art from the embodiments given herein without making any creative effort, shall fall within the protection scope of the present invention.
It should be noted that the terms "first," "second," and the like in the description and claims of the present invention and in the drawings described above are used for distinguishing between similar elements and not necessarily for describing a particular sequential or chronological order. It is to be understood that the data so used is interchangeable under appropriate circumstances such that the embodiments of the invention described herein are capable of operation in sequences other than those illustrated or described herein. Furthermore, the terms "comprises," "comprising," and "having," and any variations thereof, are intended to cover a non-exclusive inclusion, such that a process, method, system, article, or apparatus that comprises a list of steps or elements is not necessarily limited to those steps or elements expressly listed, but may include other steps or elements not expressly listed or inherent to such process, method, article, or apparatus.
The principles and advantages of the present invention are explained and illustrated below with reference to specific embodiments so that those skilled in the art may better understand the present invention. The following description is exemplary only, and is not intended to limit the scope thereof.
1. A method for rapidly detecting ethyl carbamate in white spirit based on ultra-high performance liquid chromatography combined high-resolution mass spectrometry technology comprises the following steps:
1) extracting and enriching a target object to be detected in the white spirit sample by adopting a concentration and redissolution pretreatment means to obtain a target object sample;
2) detecting the obtained target sample to be detected by using an ultra-high performance liquid chromatography-mass spectrometry technology to obtain a primary mass spectrogram and a secondary mass spectrogram of the target sample to be detected;
3) extracting target object information through software based on the obtained primary mass spectrogram and secondary mass spectrogram of the detected sample:
4) and establishing a standard spectrogram by combining with the ethyl carbamate standard substance, comparing the standard spectrogram with the obtained information of the extracted target substance, and performing qualitative and quantitative analysis on the ethyl carbamate in the white wine.
Wherein, the step 2) is specifically as follows:
2.1) detecting conditions of the ultra-high performance liquid chromatograph: chromatographic column conditions: hypersil GOLD 100X 2.1mm1.9 μm; mobile phase: 0.1% formic acid +4mM ammonium formate in water (A), 0.1% formic acid +4mM ammonium formate in methanol (B); mobile phase gradient elution procedure: 0-1min (20% B), 1-7min (100% B), 7-12min (100% B),12.1-13min (20% B), 13-15min (20% B); column oven: 30-40 ℃; sample introduction amount: 5-10 μ L; flow rate: 0.1-0.3 mL/min.
2.2) Mass Spectrometry conditions were set as: a detector: an electrostatic field orbital ion trap mass spectrometer; the scanning mode is as follows: ESI positive ion mode; scanning mode: full MS ddMS2(ii) a Scanning range m/z: 100-1500; spraying voltage: 3.5-5 kV; sheath gas pressure: 30-40 psi; auxiliary N2Pressure: 10-15 bar; auxiliary gas heating temperature: 300 ℃ and 400 ℃; capillary temperature: 300 ℃ and 350 ℃; lens voltage: 50-60V; full scan resolution: 70000 80000 FHWM; secondary scanning resolution: 17000 and 18000 FHWM; maximum residence time: primary full scan 150-: 50-100 ms; the normalized collision energy is: 17.5-52.5 eV.
Wherein, the specific operation of the step 2) is as follows: obtaining a primary mass spectrogram through a full scanning mode of a high performance liquid chromatography-ion trap orbit mass spectrometry system; and scanning by selecting an ion detection mode to obtain a secondary mass spectrogram.
Wherein, the specific operation of the step 4) is as follows: and analyzing fragmentation fragment data obtained by analyzing the secondary mass spectrogram, comparing the fragmentation fragment data with the secondary mass spectrogram of a standard substance, preliminarily judging the attribution type of the substance, and analyzing other fragment information of the retention time chromatographic peak, wherein the other fragment information comprises a mass-to-charge ratio, a fragment abundance ratio and an isotope abundance ratio, so that the structure and the element composition of the substance are deduced. And then carrying out quantification according to the primary spectrogram of the white spirit sample and the ethyl carbamate standard substance and the peak area and concentration ratio of the extracted ion-flow graph.
2. Instrument for measuring the position of a moving object
High speed refrigerated centrifuge (Beckman Coulter, usa); vortex mixer model vortex 2t, vortex mixer (Scientific Industries, usa); an ultrasonic cleaner (jiangsu kunshan ultrasonic instruments ltd); analytical balance (Mettler Toledo, switzerland); an ED-115 type constant temperature drying oven (Binder, Germany); ultra performance liquid chromatography-quadrupole-electrostatic field orbitals ion trap mass spectrometry (Thermo Fisher Scientific, usa); Milli-Q Integral model water purifier (Millipore, USA).
3. Reagent
Twelve types of wine samples (supermarkets such as Wal-Mar supermarket and Jia le Fu) with different brands and ten batches; 0.22 μm microfiltration (Pall, USA), methanol (chromatographically pure), formic acid (chromatographically pure), acetic acid (chromatographically pure), ammonium formate (chromatographically pure) (Sigma, USA); urethane Standards (LGC Standards, uk).
4. Preparation of the solution
Wherein, the specific operation of preparing the ethyl carbamate standard substance in the step 4) is as follows: urethane stock solution (5.00 mg/mL): accurately weighing 0.25g of ethyl carbamate standard substance, dissolving with ultrapure water, diluting to 50mL, and storing below 4 ℃ for later use. Urethane intermediate a (50. mu.g/mL): accurately sucking 1.00mL of the ethyl carbamate stock solution, using ultrapure water to fix the volume to 100mL, and preserving the ethyl carbamate stock solution at the temperature of below 4 ℃ for later use. Urethane intermediate b (2.5. mu.g/mL): accurately sucking 5.00mL of the urethane intermediate solution a (50 mu g/mL), diluting to 100mL with ultrapure water, and preserving at below 4 ℃ for later use.
Standard working curve solution: accurately sucking 4 mu L, 10 mu L, 20 mu L, 40 mu L and 80 mu L of the urethane intermediate solution b (2.5 mu g/mL) and 8 mu L and 20 mu L of the urethane intermediate solution a (10.0 mu g/mL) into 7 1mL volumetric flasks, and metering to a scale with ultrapure water to obtain working curve solutions of 1 mu g/L, 10 mu g/L, 25 mu g/L, 50 mu g/L, 100 mu g/L, 20 mu g/L and 400 mu g/L, which are prepared currently.
5. Sample pretreatment
Wherein, the step 1) is specifically as follows: taking 20-50mL of sample to an evaporation bottle, rotating and evaporating to dryness at 70-90 ℃, redissolving with ultrapure water, fixing the volume to 1mL, and filtering with a 0.22 mu m organic phase microporous membrane to obtain the sample to be detected.
5. Analysis results
(1) Methodology validation
a standard curve, linear range and correlation coefficient of method
And drawing a working curve according to the mass concentration corresponding to the response peak area of the ethyl carbamate in the working curve solution. The peak area of the quantitative ion chromatographic peak of the aroma component substance is expressed by the ordinate y, and the standard substance calibration curve is drawn by the concentration (mg/L) of the standard substance expressed by the x. The experimental result shows that the linear relation of the ethyl carbamate in the detection method is good and the correlation coefficient (r) is good in the concentration range of 1-400 mu g/L2) Are all greater than 0.99 (table 1).
b detection limit, quantitative limit, recovery rate and precision of the method
The detection limit is an index for measuring the sensitivity of the instrument and method, and is usually calculated by a 3-fold signal-to-noise ratio (S/N: 3) and a 10-fold signal-to-noise ratio (S/N: 10). The detection limit and the quantification limit of the experiment are respectively 0.11 mug/kg and 0.38 mug/kg. The accuracy and precision of the experiment were evaluated by the standard deviation of the recovery from the results of 6 replicates. Adding series of mixed standard substance solutions of the urethane components at high (500. mu.g/L), medium (100. mu.g/L) and low (50. mu.g/L) concentration levels into the matrix, and detecting by using a gas chromatography-mass spectrometer to calculate the average recovery rate and the relative standard deviation of various aroma flavor substances. The recovery rate and the relative standard deviation result are 81.6% -90.7% and 1.8% -5.9% respectively, and the established method has good accuracy and precision (Table 1). The experimental results show that the urethane has a good linear relationship in the linear range.
TABLE 1 urethane liquid chromatography-Mass Spectrometry parameters and methodological validation data
7. Actual sample detection
Taking 0.5-1.5mL of the sample to be detected after pretreatment in the step 2, detecting according to the detection conditions in the step 3, and obtaining a chromatogram of the sample to be detected, which is shown in a figure 1, and a chromatogram of the ethyl carbamate standard, which is shown in a figure 2. And the secondary mass spectrograms of the sample to be detected and the ethyl carbamate standard substance are shown in figures 3 and 4, and qualitative analysis is carried out on the sample to be detected. And (3) quantitatively analyzing the ethyl carbamate in the sample to be detected according to a standard curve of the ethyl carbamate, wherein detection results are lower than the limit of ethyl carbamate requirements specified in China.
In conclusion, the invention discloses a method for rapidly detecting ethyl carbamate in white spirit based on ultra-high performance liquid chromatography combined high-resolution mass spectrometry technology. Rotary evaporation is selected as a pretreatment means, high performance liquid chromatography-electrostatic field orbital ion trap mass spectrometry technology is used for analysis, and an instrument system screening method for trace ethyl carbamate in white spirit is established. The experimental result shows that the linear relation of the ethyl carbamate is good and the correlation coefficient (r) is good in the concentration range of 1-400 mu g/L2) Greater than 0.99. The recovery rate of the substrate reaches 81.6-90.7% under the three addition levels of high (500 mug/L), medium (100 mug/L) and low (50 mug/L), the experiment is repeated for 6 times, and the relative standard deviation is 1.8-5.9%. The detection limit and the quantification limit are respectively 0.11 mug kg-1With 0.38. mu.g kg-1. The method has accurate qualitative and quantitative result and high sensitivity, and provides a systematic detection method for screening the food safety risk substance ethyl carbamate in the white spirit.
The above-mentioned contents are only for illustrating the technical idea of the present invention, and the protection scope of the present invention is not limited thereby, and any modification made on the basis of the technical idea of the present invention falls within the protection scope of the claims of the present invention.
Claims (10)
1. A method for rapidly detecting ethyl carbamate in white spirit based on ultra-high performance liquid chromatography combined high resolution mass spectrometry is characterized by comprising the following steps:
1) carrying out concentration and redissolution pretreatment on the white spirit sample, and extracting and enriching a target substance to be detected in the white spirit sample to obtain a target substance to be detected sample;
2) detecting the obtained target sample to be detected based on the ultra-high performance liquid chromatography and high resolution mass spectrometry technology to obtain a primary mass spectrogram and a secondary mass spectrogram of the target sample to be detected;
3) extracting target object information through software based on the obtained primary and secondary mass spectrograms:
4) and establishing a standard spectrogram based on the ethyl carbamate standard substance, and comparing the established standard spectrogram with the obtained extracted target object information to qualitatively and quantitatively determine the ethyl carbamate in the white spirit sample.
2. The method for rapidly detecting ethyl carbamate in white spirit based on ultra-high performance liquid chromatography combined high resolution mass spectrometry as claimed in claim 1, wherein the pretreatment of concentration and redissolution in step 1) specifically comprises: and (3) putting 20-50mL of a white spirit sample into an evaporation bottle, rotating to evaporate to dryness, redissolving by ultrapure water, fixing the volume to 1mL, and finally filtering by using an organic phase microporous filter membrane to obtain a target sample to be detected.
3. The method for rapidly detecting ethyl carbamate in white spirit based on ultra-high performance liquid chromatography combined high resolution mass spectrometry as claimed in claim 2, wherein the temperature of rotary evaporation is 70-90 ℃.
4. The method for rapidly detecting ethyl carbamate in white spirit based on ultra high performance liquid chromatography combined high resolution mass spectrometry as claimed in claim 1, wherein in the step 2), the specific monitoring conditions of the ultra high performance liquid chromatograph include: chromatographic column conditions: hypersil GOLD 100X 2.1mm1.9 μm; the mobile phase A is a mixed solution of water, formic acid and ammonium formate; wherein, the volume of the formic acid accounts for 0.1 percent of the volume of the mixed solution, the concentration of the ammonium formate is 4.0mmol/L, and the rest is water; the mobile phase B is a mixed solution of methanol, formic acid and ammonium formate; wherein, the volume of the formic acid accounts for 0.1 percent of the volume of the mixed solution, the concentration of the ammonium formate is 4.0mmol/L, and the rest is methanol;
the mobile phase gradient elution procedure was: within 0-1min, the volume ratio of the mobile phase A to the mobile phase B is 80%: 20% make up the mobile phase; within 1-7min, the mobile phase B is a mobile phase; within 7-12min, the mobile phase B is a mobile phase; within 12.1-13min, the volume ratio of the mobile phase A to the mobile phase B is 80%: 20% make up the mobile phase; within 13-15min, the volume ratio of the mobile phase A to the mobile phase B is 80%: 20% make up the mobile phase;
column oven: 30-40 ℃; sample introduction amount: 5-10 μ L; flow rate: 0.1-0.3 mL/min.
5. The method for rapidly detecting ethyl carbamate in white spirit based on ultra high performance liquid chromatography combined high resolution mass spectrometry as claimed in claim 1, wherein in the step 2), the specific monitoring conditions of the high resolution mass spectrometer include: a detector: an electrostatic field orbital ion trap mass spectrometer; the scanning mode is as follows: ESI positive ion mode; scanning mode: full MS ddMS2(ii) a Scanning range m/z: 100-1500; spraying voltage: 3.5-5 kV; sheath gas pressure: 30-40 psi; auxiliary N2Pressure: 10-15 bar; auxiliary gas heating temperature: 300 ℃ and 400 ℃; capillary temperature: 300 ℃ and 350 ℃; lens voltage: 50-60V; full scan resolution: 70000 80000 FHWM; secondary scanning resolution: 17000 and 18000 FHWM; maximum residence time: primary full scan 150-: 50-100 ms; the normalized collision energy is: 17.5-52.5 eV.
6. The method for rapidly detecting ethyl carbamate in white spirit based on ultra-high performance liquid chromatography combined high resolution mass spectrometry as claimed in claim 1, wherein the specific operation of obtaining the primary mass spectrum and the secondary mass spectrum of the target sample to be detected in step 2) is as follows: and a primary mass spectrogram is obtained through a full scanning mode of a high performance liquid chromatography-ion trap orbit mass spectrum system, and a secondary mass spectrogram is obtained through scanning by selecting an ion detection mode.
7. The method for rapidly detecting ethyl carbamate in white spirit based on ultra-high performance liquid chromatography combined high resolution mass spectrometry as claimed in claim 1, wherein the specific operations of step 4) are as follows: obtaining fragmentation fragment data by analyzing the secondary mass spectrogram, comparing the fragmentation fragment data with a secondary spectrogram of an ethyl carbamate standard substance, and qualitatively judging the ethyl carbamate in the white spirit; continuously analyzing other fragment information of the retention time chromatographic peak to deduce the structure and element composition of the substance; and finally, quantifying the ethyl carbamate in the white spirit sample according to the primary spectrogram of the white spirit sample and the primary spectrogram of the ethyl carbamate standard substance and the ratio of the peak area to the concentration of the extracted ion-flow diagram.
8. The method for rapidly detecting ethyl carbamate in white spirit based on ultra-high performance liquid chromatography combined with high resolution mass spectrometry as claimed in claim 7, wherein the specific operation of analyzing other fragment information of the retention time chromatographic peak comprises: mass-to-charge ratio, fragment abundance ratio, and isotopic abundance ratio.
9. The method for rapidly detecting ethyl carbamate in white spirit based on ultra-high performance liquid chromatography combined with high resolution mass spectrometry according to claim 7, wherein the specific operations for preparing the ethyl carbamate standard in the step 4) are as follows:
preparing a stock solution of the ethyl carbamate with the concentration of 5.00mg/mL by using a standard substance of the ethyl carbamate; preparing a urethane intermediate solution a with the concentration of 50 mug/mL by using the obtained urethane stock solution; preparing a urethane intermediate solution b with the concentration of 2.5 mu g/mL by using the obtained urethane intermediate solution a; respectively sucking the urethane intermediate liquid b: 4. mu.L, 10. mu.L, 20. mu.L, 40. mu.L, 80. mu.L and urethane intermediate liquid a: mu.L and 20. mu.L were put in 7 volumetric flasks and the volume was adjusted to 1mL to obtain 10. mu.g/L, 25. mu.g/L, 50. mu.g/L, 100. mu.g/L, 20. mu.g/L and 400. mu.g/L curve working solutions.
10. The method for rapidly detecting ethyl carbamate in white spirit based on ultra-high performance liquid chromatography combined high resolution mass spectrometry as claimed in claim 9, wherein the ethyl carbamate stock solution, the ethyl carbamate intermediate solution a and the ethyl carbamate intermediate solution b are stored at a temperature below 4 ℃ for later use; the curve working solution needs to be prepared at present.
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