CN112595794A - Method for rapidly detecting residual amount of hexadienoeestrol in milk and milk powder - Google Patents
Method for rapidly detecting residual amount of hexadienoeestrol in milk and milk powder Download PDFInfo
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Abstract
The invention discloses a method for rapidly detecting the residual amount of hexadiene estrol in milk and milk powder, which carries out HPLC-MS/MS detection after pretreatment on a sample to be detected: the target compound is directly tested on a machine after acidified acetonitrile extraction, QuEChERS technology purification and redissolution, and the pretreatment operation is simple. The optimized method has low matrix effect and simplified experimental steps. The method has high specificity, sensitivity and stability, is simple to operate, and is more suitable for large-batch detection.
Description
Technical Field
The invention relates to the technical field of veterinary drug residue detection, and particularly relates to a liquid chromatography-tandem mass spectrometry detection method for rapidly detecting the residue amount of dienestrol in a dairy product.
Background
Milk and products thereof are one of important foods in life, and are favored by people because of being rich in protein, vitamins, calcium and other nutrients required by human bodies. Dienestrol (DIE) is an artificially synthesized hormone, and is prohibited from being used in many countries due to the harm to human bodies and the environment. The national people's republic of China agricultural Ministry of rural Notification No. 250 also lists hexadienoic estradiol as a list of drugs and other compounds that are banned for use in food animals.
The existing detection method GB/T22992-2008 'determination of residual quantity of zearalanol, zearalenone, diethylstilbestrol and dienestrol in milk and milk powder-liquid chromatography-tandem mass spectrometry' has relatively complex pretreatment process, needs MAX solid phase extraction column purification, and has long time consumption of the pretreatment process and high material cost. Especially for products with large demand, such as milk and milk powder, a large number of detection samples are obtained, and how to shorten the detection time and improve the detection efficiency is a key factor for saving the cost. Therefore, the development of a method for rapidly and efficiently detecting the residual amount of the hexadienoeestrol in the milk and the milk powder has very important significance for guaranteeing the food safety of people.
Disclosure of Invention
In order to overcome the defects in the prior art, the invention provides a method for rapidly detecting the residue of hexadiene estrol in milk and milk powder. The method has the advantages of high sensitivity, high stability, simple operation and lower detection limit.
In order to achieve the purpose, the invention adopts the following technical scheme:
a method for rapidly detecting hexadienoeestrol residue in milk and milk powder comprises the following steps:
(1) preparing a standard solution: firstly, preparing a standard stock solution, weighing a proper amount of hexadienoeestrol standard substance, preparing the standard stock solution with the concentration of 10 mu g/L by using methanol, and storing the standard stock solution in a refrigerator at the temperature of 18 ℃ below zero; then preparing the standard stock solution into mixed standard working solution with the concentration of 1000 mug/mL by using methanol, and storing the mixed standard working solution in a refrigerator at 4 ℃; then preparing the standard intermediate solution into a mixed standard working solution with the concentration of 10 mug/mL by using methanol, and storing the mixed standard working solution in a refrigerator at the temperature of 4 ℃;
when a standard curve is made, preparing standard working solution into standard substance solutions with the concentrations of 1ng/mL, 5ng/mL, 10ng/mL, 15ng/mL and 20ng/mL respectively by using a blank matrix solution for later use;
(2) acetic acid acetonitrile solution: sucking 10ml of acetic acid, and using acetonitrile to fix the volume to a volumetric flask of 1000ml to prepare an acetic acid acetonitrile solution with the concentration of 1%;
(3) 10% aqueous methanol (containing 0.1% formic acid): adding 10ml of methanol into water to fix the volume to 100ml, adding 100 mul of formic acid to prepare 10 percent methanol water 0.1 percent formic acid solution;
(4) pretreatment of a sample to be detected:
A. extraction: weighing 2g (accurate to 0.01g) of milk or milk powder into a 50mL centrifuge tube, and adding 10 μ L of 100ng/mL hexadienoerine estradiol standard solution and 15mL of 1% acetonitrile acetate solution (adding 10mL of n-hexane to a sample containing more grease) during recovery. Mixing for 5min by vortex, extracting with ultrasound for 15min, mixing for 5min by vortex, and centrifuging for 5min in a centrifuge at rotation speed of 4000 r/min;
B. purifying: the whole amount of the supernatant (the n-hexane was removed from the n-hexane-added sample) was transferred to a clean tube (10g of muffle-dried anhydrous sodium sulfate, 0.5g C)180.2g PSA), evenly mixing for 5min by vortex, and centrifuging for 5min in a centrifugal machine with the rotating speed of 4000 r/min. Sucking 7.5mL of supernatant into a 10mL centrifuge tube, and drying by nitrogen in a water bath at 40 ℃;
C. redissolving: adding 0.5mL of a fixed solution (10% methanol water contains 0.1% formic acid) into the blow-dried centrifuge tube, performing ultrasonic treatment for 30s, performing vortex mixing for 5min, filtering with a 0.22 mu m filter membrane, and waiting for testing on a machine;
(5) quantitatively analyzing the pretreated sample by using a liquid chromatography-tandem mass spectrometer, and qualitatively and quantitatively detecting the sample to be detected to obtain the content of the residual hexadienoic acid in milk and milk powder, wherein the content is as follows:
A. analyzing and detecting the hexadiene estrol residual standard substance solutions with different concentrations prepared in the step (1) by using a liquid chromatogram-tandem mass spectrometer under the same detection condition as that of a sample solution to be detected, and obtaining a total ion current chromatogram, a quantitative ion chromatogram and a qualitative ion pair chromatogram of the standard substance solution;
B. sampling 5.0 mu L of sample solution to be detected, and analyzing and detecting by using a liquid chromatography-tandem mass spectrometer to obtain a total ion current chromatogram, a quantitative ion pair chromatogram and a qualitative ion pair chromatogram of the sample solution to be detected;
C. preparing a hexadiene estrol residue standard curve by using chromatographic peaks of quantitative ion pairs and qualitative ion pairs of the hexadiene estrol residue;
D. according to the chromatographic peaks of the quantitative ion pair and the qualitative ion pair of the hexadiene estrol residue in the sample solution to be detected, the concentration C of the hexadiene estrol residue in the sample solution to be detected is calculated by combining a standard curve, and the content X of the hexadiene estrol residue in milk and milk powder is calculated according to the following formula, wherein the content calculation formula is as follows:
X＝C*V*f/(M*1000)
wherein X is the content of the residue of the milk powder and the milk hexadiene estrol in the sample to be detected, and the unit is mu g/kg; c is the concentration of the residual hexadiene estrol in the sample to be detected, and the unit is ng/mL; v is the dissolving volume of the sample to be detected, and the unit is mL; m is the mass of the sample to be detected, and the unit is g; f is the dilution factor.
Wherein the chromatographic conditions of the liquid chromatogram-tandem mass spectrometer are as follows:
liquid chromatography conditions:
a) a chromatographic column: rapid Resolution HD 3.0X100mm 1.8.8-Micron;
b) mobile phase and gradient: acetonitrile + 0.1% formic acid 5mM ammonium acetate;
TABLE 1 gradient elution of mobile phase
c) End time: 5 min;
d) flow rate: 0.4 mL/min;
e) column temperature: 30 ℃;
f) sample introduction amount: 5 μ L.
Mass spectrum/mass spectrum conditions: see table 2, table 3.
TABLE 2 Agilent 6460QQQ
TABLE 3 hexadienoic Estrol monitoring ion pairs and their corresponding collision energies
The invention has the beneficial effects that:
(1) the pretreatment process is purified by using the QuECHERS technology, compared with the existing detection method, the operation is simpler and quicker, and the detection efficiency is improved.
(2) The method simplifies the detection process of the hexadiene estrol in milk and milk powder by adjusting various parameters in the prior detection technology, particularly optimizing the solvent, conditions and the like adopted by the sample pretreatment during the sample pretreatment. In the extraction step, acidified acetonitrile with the concentration of 1% is used for extraction, and compared with the method of simply using acetonitrile, the recovery rate is obviously improved.
(3) The detection method can detect the residual amount of the hexadiene estrol in the milk and the milk powder, the standard recovery rate is 80-120%, the detection limit is 0.5 mug/kg, and the sensitivity is higher.
(4) The invention optimizes and selects anhydrous sodium sulfate and anhydrous sodium sulfate C18The PSA three substances are used as purification fillers, the proportion of the purification fillers is optimized (the mass ratio of anhydrous sodium sulfate to C18 to PSA is 10 g: 0.5 g: 0.2g), the high-purity and high-controllable specific surface area ensures stable extraction efficiency, and compared with the traditional detection method, the method has the advantages of simple operation, good reproducibility and high recovery rate; compared with the use of other fillers, the cost of instruments and consumables can be greatly saved.
Drawings
FIG. 1 is a total ion flow diagram of a standard sample of dienestrol in a sample solution;
FIG. 2 is a total ion flow diagram of a standard sample of dienestrol in a standard solution;
FIG. 3 is a signal-to-noise ratio spectrum of dienestol in a sample solution with detection limit concentration added and recovered;
FIG. 4 is a standard curve for dienestol;
FIG. 5 is a quantitative qualitative ion pair chromatogram of dienestrol in a sample solution;
FIG. 6 is a quantitative qualitative ion pair chromatogram of dienestrol in a standard control solution.
Detailed Description
The invention will be further explained by means of specific embodiments, however, it should be understood that the invention may be embodied in various forms and should not be limited by the embodiments set forth herein. Rather, these embodiments are provided so that this disclosure will be thorough and complete, and will fully convey the scope of the invention to those skilled in the art.
In the following description and in the claims, the terms "include" and "comprise" are used in an open-ended fashion, and thus should be interpreted to mean "include, but not limited to. The description which follows is a preferred embodiment of the invention, but is made for the purpose of illustrating the general principles of the invention and not for the purpose of limiting the scope of the invention. The scope of the present invention is defined by the appended claims.
First, selected reagent
1. Hexadiene estrol standard substance
2. Glacial acetic acid
3. Acetonitrile
4. N-hexane
5. Anhydrous sodium sulfate
6. Methanol
7. Formic acid
8.C18：50μm 
9.PSA：40-60μm
Second, the used instrument
1. Centrifuge tube, 2, nitrogen-blown appearance: N-EVA p 112; 3. a centrifuge: SIGMA 3K15, 4, a bottle mouth liquid distributor (10-50 mL), 5, a liquid transferring gun, 6 and a vortex oscillator; heidolph MULTI ream, 7, liquid chromatography/tandem mass spectrometer (equipped with electrospray ion source): agilent 1290+6460QQQ, 8, one hundred thousand balance: METTER TOLEDO XS205, 9, one ten thousandth balance: METTER TOLEDO ME204E
Third, test
(1) Preparing a standard solution: firstly, preparing a standard stock solution, weighing a proper amount of hexadienoeestrol standard substance, preparing the standard stock solution with the concentration of 10 mu g/L by using methanol, and storing the standard stock solution in a refrigerator at the temperature of 18 ℃ below zero; then preparing the standard stock solution into a mixed standard intermediate solution with the concentration of 1000 mug/mL by using methanol, and storing the mixed standard intermediate solution in a refrigerator at 4 ℃; then preparing the standard intermediate solution into a mixed standard working solution with the concentration of 10 mug/mL by using methanol, and storing the mixed standard working solution in a refrigerator at the temperature of 4 ℃;
sucking a proper amount of mixed standard substance control solution according to experiment needs, preparing standard substance working curves with the concentrations of 1ng/mL, 5ng/mL, 10ng/mL, 15ng/mL and 20ng/mL by using the blank sample matrix solution respectively, and preparing the standard substance working curves on site when the standard substance working curves are used;
(2) acetic acid acetonitrile solution: sucking 10ml of acetic acid, and using acetonitrile to fix the volume to a volumetric flask of 1000ml to prepare an acetic acid acetonitrile solution with the concentration of 1%;
(3) aqueous methanol (containing 0.1% formic acid) solution: adding 10ml of methanol into water to fix the volume to 100ml, adding 100 mul of formic acid to prepare 10 percent methanol water 0.1 percent formic acid solution;
(4) pretreatment of a sample to be detected:
A. extraction: weighing 2g (accurate to 0.01g) of milk or milk powder into a 50mL centrifuge tube, and adding 10 μ L of 100ng/mL hexadienoerine estradiol standard solution and 15mL of 1% acetonitrile acetate solution (adding 10mL of n-hexane to a sample containing more grease) during recovery. Mixing for 5min by vortex, extracting with ultrasound for 15min, mixing for 5min by vortex, and centrifuging for 5min in a centrifuge at rotation speed of 4000 r/min;
B. purifying: the whole amount of the supernatant (the n-hexane was removed from the n-hexane-added sample) was transferred to a clean tube (10g of muffle-dried anhydrous sodium sulfate, 0.5g C)180.2g PSA) (see tables 4-6), vortex and mix evenly for 5min, centrifuge for 5min in a centrifuge with the rotation speed of 4000 r/min. Aspirate 7.5mL of supernatant into a 10mL centrifuge tube at 40 deg.CDrying with nitrogen in water bath.
C. Redissolving: adding 0.5mL of a fixed solution (10% methanol water contains 0.1% formic acid), performing ultrasonic treatment for 30S, performing vortex mixing for 5min, filtering with a 0.22um filter membrane, and performing machine test.
Blank matrix solution the blank matrix solution was run as in a.b.c, except that no standard solution was added.
(5) Quantitatively analyzing the pretreated sample liquid by using a liquid chromatography-tandem mass spectrometer, and carrying out qualitative and quantitative detection on a sample to be detected to obtain the content of the residual hexadienyl estrol in milk and milk powder, wherein the content is as follows:
A. analyzing and detecting the hexadienyl estrol standard working curves with different concentrations prepared in the step (1) by using a liquid chromatography-tandem mass spectrometer, wherein the sample injection volume is 5.0 mu L, and obtaining a total ion flow graph, a quantitative ion chromatogram and a qualitative ion pair chromatogram of a standard solution;
B. sampling 5.0 mu L of sample solution to be detected, and analyzing and detecting by using a liquid chromatography-tandem mass spectrometer to obtain a total ion flow graph, a quantitative ion pair chromatogram and a qualitative ion pair chromatogram of the sample solution to be detected;
C. respectively taking the quantitative ion peak area and the concentration of the hexadiene estrol as horizontal and vertical coordinates to obtain a standard working curve of the hexadiene estrol;
D. according to the quantitative ion peak area of the residue of the hexadiene estrol in the sample solution to be detected, the concentration C of the hexadiene estrol in the sample solution to be detected is calculated by combining a standard curve, and the content X of the hexadiene estrol in milk and milk powder is calculated according to the following formula, wherein the content calculation formula is as follows:
X＝C*V*1000*f/(m*1000)
wherein X is the content of milk powder and milk hexadienyl estrol in a sample to be detected, and the unit is mu g/kg; c is the concentration of the residual hexadiene estrol in the sample to be detected, and the unit is ng/mL; v is the final constant volume of the sample to be measured, and the unit is mL; m is the mass of the sample to be detected and the unit is g; f is the dilution factor.
The mass spectrum conditions of the liquid chromatogram-mass spectrum combination instrument are as follows:
ionization mode: electrospray ionization negative ion mode; the scanning mode is as follows: multiple Reaction Monitoring (MRM); ejection voltage: 2000V; delta EM: 200V; atomizer pressure: 40 psi; flow rate of drying gas: 10L/min; temperature of the drying gas: 345 deg.C.
Fourth, process optimization and analysis
The process of the invention carries out optimization test on the selection proportion of the extraction solvent and the purification filler respectively, and finally selects the extraction solvent as follows according to the detected recovery rate condition: 1% acetonitrile acetate, the filler and the proportion are: anhydrous sodium sulfate + C18+PSA＝10g+0.5g+0.2g。
The specific comparison results are shown in tables 4-7.
(1) Optimization of extraction solvent selection
The acetonitrile used in the conventional method and the acidified acetonitrile with different concentration gradients are selected to extract the positive sample, the recovery rate is determined, and the following conditions can be seen in the table 4: the extraction effect of the 1% acetonitrile acetate is better.
TABLE 4 optimization of the extraction solvent ratio
| Extraction solvent | Percent recovery% |
| Acetonitrile | 95.53 |
| 1% acetonitrile acetate | 98.64 |
| 5% acetonitrile acetate | 95.22 |
| 10% acetonitrile acetate | 95.54 |
(1) Purification packing selection optimization
Currently, the common purification fillers for the QuEChERS technology are: anhydrous magnesium sulfate, anhydrous sodium sulfate, sodium chloride, C18PSA, graphitized carbon black, alumina, and the like. Combining the components of the milk and milk powder to be tested, selecting anhydrous sodium sulfate to remove water, and PSA (N-propyl ethylenediamine) to remove fatty acid, organic acid, and some polar pigment and sugar. C18The functional group has adsorption effect on nonpolar components, and can remove oil and fat substances. Optimizing the selected filler proportion, measuring the recovery rate, counting as follows, and finally selecting the proportion with the best recovery effect: sodium sulfate + C18+ PSA ═ 10g +0.5g +0.2g as final protocol.
TABLE 5 purification tube Filler C18Optimizing table
| Anhydrous sodium sulfate | C18 | PSA | Recovery rate |
| 10g | 0.2g | 0.2g | 93.33 |
| 10g | 0.5g | 0.2g | 97.56 |
| 10g | 1.0g | 0.2g | 96.28 |
| 10g | 2.0g | 0.2g | 96.23 |
TABLE 6 purifying tube packing PSA optimizing table
| Anhydrous sodium sulfate | C18 | PSA | Recovery rate |
| 10g | 0.5g | 0.1g | 89.43 |
| 10g | 0.5g | 0.2g | 96.58 |
| 10g | 0.5g | 0.5g | 94.28 |
| 10g | 0.5g | 1.0g | 94.19 |
TABLE 7 optimization table for anhydrous sodium sulfate as filler of purification tube
| Anhydrous sodium sulfate | C18 | PSA | Recovery rate |
| 5g | 0.5g | 0.2g | 91.25 |
| 10g | 0.5g | 0.2g | 96.60 |
| 15g | 0.5g | 0.2g | 94.35 |
| 20g | 0.5g | 0.2g | 95.31 |
Fifth, testing and analyzing
(1) Detection limit
The detection limit level (0.5 mug/kg) is selected for carrying out the addition recovery test, the signal-to-noise ratio S/N of the method is 1173.1, as shown in figure 3, when the signal-to-noise ratio is calculated to be equal to 10, the corresponding concentration is that the quantitative limit of the method can be as low as 0.004 mug/kg.
(2) Linearity of the standard curve
The measurement was carried out according to the above-mentioned detection method using a standard working curve prepared with a blank medium at a concentration of 1ng/mL, 5ng/mL, 10ng/mL, 15ng/mL, or 20 ng/mL.
The linear correlation coefficient of the standard curve of the hexadienoeestrol is more than 0.99 and meets the requirement obtained from the figure 4.
(3) Recovery rate and accuracy
For this substance, the measurement lower limit, the two-fold method measurement lower limit and the ten-fold method measurement lower limit were selected for three-level addition, and the number of repeated measurements was 6, that is, the present invention performed the labeling measurement at the levels of 0.5. mu.g/kg, 1. mu.g/kg and 5. mu.g/kg, respectively, and the samples were tested, and the results of the measurements are shown in Table 8.
TABLE 8 results of recovery and accuracy measurements
As can be seen from the table, the recovery rates of the three different levels of addition recovery tests are all over 90 percent, the recovery rates meet the requirements, the same level is subjected to 6 times of parallel measurement, the calculated relative standard deviation is less than 10 percent, and the repeatability of the method meets the requirements.
The above description is only for the purpose of illustrating the preferred embodiments of the present invention and is not to be construed as limiting the invention, as any modification, equivalent replacement, simple improvement or the like made in the spirit of the present invention should be included in the present invention.
Claims (7)
1. A method for rapidly detecting the residual amount of hexadienoeestrol in milk and milk powder is characterized by comprising the following steps:
(1) preparing a standard solution: firstly, preparing a standard stock solution, weighing a proper amount of hexadienoeestrol standard substance, preparing the standard stock solution with the concentration of 10 mu g/L by using methanol, and storing the standard stock solution in a refrigerator at the temperature of 18 ℃ below zero; then preparing the standard stock solution into a mixed standard intermediate solution with the concentration of 1000 mug/mL by using methanol, and storing the mixed standard intermediate solution in a refrigerator at 4 ℃; then preparing the standard intermediate solution into a mixed standard working solution with the concentration of 10 mug/mL by using methanol, and storing the mixed standard working solution in a refrigerator at the temperature of 4 ℃;
when a standard curve is made, preparing standard working solution into standard substance solutions with the concentrations of 1ng/mL, 5ng/mL, 10ng/mL, 15ng/mL and 20ng/mL respectively by using a blank matrix solution for later use;
(2) pretreatment of a sample to be detected:
A. extraction: weighing 2.00g of milk or milk powder into a 50mL centrifuge tube, and adding 10 mu L of 1000ng/mL hexadienyl estradiol standard solution and 15mL of 1% acetic acid acetonitrile solution in the recovery process; mixing for 5min by vortex, extracting with ultrasound for 15min, mixing for 5min by vortex, and centrifuging for 5min in a centrifuge at rotation speed of 4000 r/min;
B. purifying: the supernatant was transferred in its entirety to a prepared purification tube containing 10g of muffle-dried anhydrous sodium sulfate, 0.5g C180.2g of PSA, uniformly mixing for 5min in a vortex manner, centrifuging for 5min in a centrifuge with the rotating speed of 4000r/min, sucking 7.5mL of supernatant into a 10mL centrifuge tube, and drying in a water bath at 40 ℃ by using nitrogen;
C. redissolving: adding 0.5mL of 10% methanol water containing 0.1% formic acid into the dried centrifuge tube, performing ultrasonic treatment for 30S, performing vortex mixing for 5min, filtering with a 0.22 μm filter membrane, and waiting for testing on a machine.
(3) And (3) quantitatively analyzing the pretreated sample by using a liquid chromatography-tandem mass spectrometer, and qualitatively and quantitatively detecting the sample to be detected to obtain the content of the residual hexadienoic acid in the milk and the milk powder.
2. The method for detecting the residual amount of hexadienoeestrol in milk and milk powder according to claim 1, wherein the step (3) is as follows:
A. analyzing and detecting the hexadienyl estrol standard working curves with different concentrations prepared in the step (1) by using a liquid chromatography-tandem mass spectrometer, wherein the sample injection volume is 5.0 mu L, and obtaining a total ion flow graph, a quantitative ion chromatogram and a qualitative ion pair chromatogram of a standard solution;
B. sampling 5.0 mu L of sample solution to be detected, and analyzing and detecting by using a liquid chromatography-tandem mass spectrometer to obtain a total ion flow graph, a quantitative ion pair chromatogram and a qualitative ion pair chromatogram of the sample solution to be detected;
C. respectively taking the quantitative ion peak area and the concentration of the hexadiene estrol as horizontal and vertical coordinates to obtain a standard working curve of the hexadiene estrol;
D. according to the quantitative ion peak area of the residue of the hexadiene estrol in the sample solution to be detected, the concentration C of the hexadiene estrol in the sample solution to be detected is calculated by combining a standard curve, and the content X of the hexadiene estrol in milk and milk powder is calculated according to the following formula, wherein the content calculation formula is as follows:
X＝C*V*1000*f/(m*1000)
wherein X is the content of milk powder and milk hexadienyl estrol in a sample to be detected, and the unit is mu g/kg; c is the concentration of the residual hexadiene estrol in the sample liquid to be detected, and the unit is ng/mL; v is the final constant volume of the sample to be measured, and the unit is mL; m is the mass of the sample to be detected and the unit is g; f is the dilution factor.
3. The method for detecting the residual amount of hexadienoeestrol in milk and milk powder according to claim 1, wherein the chromatographic conditions of the liquid chromatography-tandem mass spectrometer are as follows:
a) chromatographic column Rapid Resolution HD 3.0X100mm 1.8.8-Micron;
b) mobile phase and gradient: acetonitrile + 0.1% formic acid 5mM ammonium acetate;
c) end time: 5 min;
d) flow rate: 0.4 mL/min;
e) column temperature: 30 ℃;
f) sample introduction amount: 5 μ L.
4. The method for detecting the residual amount of hexadienoeestrol in milk and milk powder according to claim 3, wherein the volume ratio of the acetonitrile to the ammonium acetate is (20-90): (10-80).
5. The method for detecting the residual amount of dienestrol in milk and milk powder according to claim 3, wherein the gradient elution is specified in the following table:
TABLE 1 gradient elution of mobile phase
6. The method for detecting the residual amount of the hexadienoeestrol in the milk and the milk powder according to claim 3, wherein the mass spectrum conditions of the liquid chromatography-mass spectrometer are as follows: ionization mode: electrospray ionization negative ion mode; the scanning mode is as follows: multiple Reaction Monitoring (MRM); ejection voltage: 4000V; delta EM: 200V; atomizer pressure: 40 psi; flow rate of drying gas: 7L/min; temperature of the drying gas: 325 ℃.
7. The method for detecting the residual amount of dexamethasone in milk and milk powder as claimed in claim 1, wherein in the extraction process of step (2), for the sample with more oil and fat, in addition to 15mL of 1% acetonitrile acetate solution, 10mL of n-hexane is added, and in the purification process of step (2), the n-hexane is removed from the sample with n-hexane added.
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