CN112067724A - Qualitative detection method for soluble sugar acid in pear fruit - Google Patents
Qualitative detection method for soluble sugar acid in pear fruit Download PDFInfo
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Abstract
The invention provides a qualitative detection method of soluble sugar acid in pear fruits, and belongs to the technical field of organic detection. The method has the advantages that the soluble sugar is detected by ultra-high performance liquid chromatography (UPLC), the qualitative detection of the soluble sugar and the organic acid is realized by accurately limiting the conditions of the ultra-high performance liquid chromatography, the determination of the soluble sugar in the method can be controlled within 14min to be complete, the determination of the organic acid can be controlled within 5min to be complete, the time is saved compared with the conventional High Performance Liquid Chromatography (HPLC) (the soluble sugar is peak within 20min, and the organic acid is peak within 15 min), the automation degree is high, the repeatability is good, the method can be popularized to the research of the sugar acid content and the components of fruits in the future, and the large-batch detection of the sugar acid content in the pear fruits can be realized.
Description
Technical Field
The invention relates to the technical field of organic detection, in particular to a qualitative detection method for soluble sugar acid in pear fruits.
Background
Pears belong to the Rosaceae (Rosaceae) and Pyrioideae (Pomaceae) genus (Pyrus) plants, and are one of the important deciduous fruit trees in the world. In our country, the pear is the third largest fruit after apples and citrus. The sugar acid is an important component of the internal quality of the fruit, and the variety, the content and the proportion of the sugar acid play a decisive role in the commodity value, the taste and the flavor of the fruit, are concerned by people all the time and are one of important indexes for the researchers to improve all the time. The research on the saccharic acid content and components in the pear fruit is helpful for understanding the formation factors of the internal quality of the pear fruit and the difference of the saccharic acid content and the components among different varieties, so that parents with excellent quality are screened out, and the breeding process is accelerated.
The existing pear sugar acid qualitative detection methods comprise an anthrone colorimetric method, a film reagent colorimetric method, a high-efficiency pore tube electrophoresis method, a dinitrosalicylic acid method, a sodium hydroxide titration method and a High Performance Liquid Chromatography (HPLC), but the existing pear sugar acid qualitative detection methods have the problem of long detection time.
Disclosure of Invention
In view of the above, the present invention aims to provide a method for qualitatively detecting soluble sugar acid in pear fruit. The qualitative detection method provided by the invention can realize rapid qualitative determination of specific types of soluble sugar acids.
In order to achieve the above object, the present invention provides the following technical solutions:
the invention provides a qualitative detection method of soluble sugar acid in pear fruits, wherein the sugar comprises fructose, sorbitol, glucose and sucrose, the acid is organic acid, the organic acid comprises oxalic acid, quinic acid, malic acid, shikimic acid and citric acid, and the method comprises the following steps:
mixing a pear fruit sample, water and a grinding medium, and then sequentially grinding, leaching, heating and naturally cooling to room temperature to obtain a liquid to be detected;
performing first ultra-high performance liquid chromatography detection on the solution to be detected to obtain the species of soluble sugar in the pear fruit sample, wherein the conditions of the first ultra-high performance liquid chromatography detection comprise:
the high-efficiency liquid phase conditions include: a chromatographic column: UPLCACQUITYBEHAmide; mobile phase: the acetonitrile-ammonia water mixed solution and water are mixed according to the volume ratio of 85:15, the mass fraction of ammonia water in the acetonitrile-ammonia water mixed solution is 1%, and the flow rate is as follows: 0.2 mL/min-1Column temperature: 45 ℃, sample injection time: 15min, sample injection volume: 2 mu L of the solution;
the detection conditions include: ELSD detector, gas pressure: 25Psi, drift tube temperature: 55 ℃, sprayer: 25 ℃;
and carrying out second ultra high performance liquid chromatography detection on the liquid to be detected to obtain the species of the organic acid in the pear fruit sample, wherein the conditions of the second ultra high performance liquid chromatography detection comprise:
the high-efficiency liquid phase conditions include: a chromatographic column: ACQUITYUPLCHSST 3; mobile phase: potassium dihydrogen phosphate solution-methanol, wherein the concentration of potassium dihydrogen phosphate in the mobile phase is 20mmol/L, the mass fraction of methanol is 1%, the pH value of the mobile phase is 2.4, and the flow rate is as follows: 0.25 mL/min-1Column temperature: 30 ℃, sample injection time: 5min, sample introduction volume：2μL；
The detection conditions include: PDA detector, wavelength: 214 nm.
Preferably, the dosage ratio of the pear fruit sample to water is 0.5-1.0 g: 2-4 mL.
Preferably, the grinding medium is ceramic beads, and the particle size of the ceramic beads is 2-3 mm.
Preferably, the rotation speed of the grinding is 6-8 m.s-1The time is 45-60 s.
Preferably, the leaching is carried out under ultrasonic conditions, and the leaching time is 15-20 min.
Preferably, the heating temperature is 80-90 ℃, and the heating heat preservation time is 30-35 min.
Preferably, after naturally cooling to room temperature, the method further comprises: centrifuging the obtained cooling system to obtain a supernatant; and filtering the supernatant by using a 0.22 mu m water system filter membrane, wherein the filtrate is the liquid to be detected.
Preferably, the temperature of the centrifugation is 4 ℃, and the rotating speed is 12000-14000 r.min-1The time is 15-20 min.
Preferably, the specification of the UPLCACQUITYBEHAmide chromatographic column is 2.1mm × 100mm × 1.7 μm.
Preferably, the ACQUITYUPLCHSST3 column has a specification of 100mm × 2.1mm × 1.8 μm.
The invention provides a qualitative detection method of soluble sugar acid in pear fruit, wherein the sugar comprises fructose, sorbitol, glucose and sucrose, the acid is organic acid, the organic acid comprises oxalic acid, quinic acid, malic acid, shikimic acid and citric acid, and the method comprises the following steps: mixing a pear fruit sample, water and a grinding medium, and then sequentially grinding, leaching, heating and naturally cooling to room temperature to obtain a liquid to be detected; performing first ultra-high performance liquid chromatography detection on the solution to be detected to obtain the species of soluble sugar in the pear fruit sample, wherein the conditions of the first ultra-high performance liquid chromatography detection comprise: the high-efficiency liquid phase conditions include: a chromatographic column: UPLCACQUITYBEHAmide; mobile phase: the acetonitrile-ammonia water mixed solution and water are mixed according to the volume ratio of 85:15, and the acetonitrile-ammonia waterThe mass fraction of ammonia water in the mixed solution is 1%, and the flow rate is as follows: 0.2 mL/min-1Column temperature: 45 ℃, sample injection time: 15min, sample injection volume: 2 mu L of the solution; the detection conditions include: ELSD detector, gas pressure: 25Psi, drift tube temperature: 55 ℃, sprayer: 25 ℃; and carrying out second ultra high performance liquid chromatography detection on the liquid to be detected to obtain the species of the organic acid in the pear fruit sample, wherein the conditions of the second ultra high performance liquid chromatography detection comprise: the high-efficiency liquid phase conditions include: a chromatographic column: ACQUITYUPLCHSST 3; mobile phase: potassium dihydrogen phosphate solution-methanol, wherein the concentration of potassium dihydrogen phosphate in the mobile phase is 20mmol/L, the mass fraction of methanol is 1%, the pH value of the mobile phase is 2.4, and the flow rate is as follows: 0.25 mL/min-1Column temperature: 30 ℃, sample injection time: 5min, sample injection volume: 2 mu L of the solution; the detection conditions include: PDA detector, wavelength: 214 nm.
The invention has the following beneficial effects:
the invention provides a rapid qualitative determination method for soluble sugar and organic acid in pear fruits, the determination of the soluble sugar and the organic acid in the invention adopts Ultra Performance Liquid Chromatography (UPLC) detection, the invention realizes the qualitative detection of the soluble sugar and the organic acid by accurately limiting the conditions of the ultra performance liquid chromatography detection, and the determination of the soluble sugar in the method can be controlled to be complete within 14min, and the determination of the organic acid can be controlled to be complete within 5min, thus saving time compared with the conventional High Performance Liquid Chromatography (HPLC) (the soluble sugar is complete within 20min, and the organic acid is complete within 15 min), having high automation degree and good repeatability, being capable of being popularized to the research of the sugar acid content and components of the pear fruits in the future, and realizing the large-scale detection of the sugar acid content in the pear fruits; compared with the conventional sugar acid extraction method (the time for using a single sample is 3.5 hours), the method saves more time, and the conventional ethanol solution is changed into water in the extraction process, so that the use of organic solvents is reduced, and the safety of experiments is improved.
Drawings
FIG. 1 is a peak profile of a soluble sugar mixed standard;
FIG. 2 is a peak-appearing diagram of an organic acid mixed standard;
FIG. 3 is a graph of soluble sugar peaks in a pear pulp sample;
fig. 4 is a graph of the peak appearance of organic acids in a pear pulp sample.
Detailed Description
The invention provides a qualitative detection method of soluble sugar acid in pear fruits, wherein the sugar comprises fructose, sorbitol, glucose and sucrose, the acid is organic acid, the organic acid comprises oxalic acid, quinic acid, malic acid, shikimic acid and citric acid, and the method comprises the following steps:
mixing a pear fruit sample, water and a grinding medium, and then sequentially grinding, leaching, heating and naturally cooling to room temperature to obtain a liquid to be detected;
performing first ultra-high performance liquid chromatography detection on the solution to be detected to obtain the species of soluble sugar in the pear fruit sample, wherein the conditions of the first ultra-high performance liquid chromatography detection comprise:
the high-efficiency liquid phase conditions include: a chromatographic column: UPLCACQUITYBEHAmide; mobile phase: the acetonitrile-ammonia water mixed solution and water are mixed according to the volume ratio of 85:15, the mass fraction of ammonia water in the acetonitrile-ammonia water mixed solution is 1%, and the flow rate is as follows: 0.2 mL/min-1Column temperature: 45 ℃, sample injection time: 15min, sample injection volume: 2 mu L of the solution;
the detection conditions include: ELSD detector, gas pressure: 25Psi, drift tube temperature: 55 ℃, sprayer: 25 ℃;
and carrying out second ultra high performance liquid chromatography detection on the liquid to be detected to obtain the species of the organic acid in the pear fruit sample, wherein the conditions of the second ultra high performance liquid chromatography detection comprise:
the high-efficiency liquid phase conditions include: a chromatographic column: ACQUITYUPLCHSST 3; mobile phase: potassium dihydrogen phosphate solution-methanol, wherein the concentration of potassium dihydrogen phosphate in the mobile phase is 20mmol/L, the mass fraction of methanol is 1%, the pH value of the mobile phase is 2.4, and the flow rate is as follows: 0.25 mL/min-1Column temperature: 30 ℃, sample injection time: 5min, sample injection volume: 2 mu L of the solution;
the detection conditions include: PDA detector, wavelength: 214 nm.
The method comprises the steps of mixing a pear fruit sample, water and a grinding medium, and then sequentially grinding, leaching, heating and naturally cooling to room temperature to obtain a liquid to be detected. The source of the pear fruit sample is not particularly limited in the invention, and the method is well known to those skilled in the art.
In the invention, the dosage ratio of the pear fruit sample to water is preferably 0.5-1.0 g: 2-4 mL, more preferably 0.6-0.8 g: 3 mL. In the present invention, the water is preferably ultrapure water. In the invention, the pear fruit sample is preferably ground into powder under the condition of liquid nitrogen and then mixed with water and grinding media. In the present invention, the rotation speed of the grinding is preferably 6m/s, and the time is preferably 45 s.
In the invention, the grinding medium is preferably ceramic beads, the particle size of the ceramic beads is preferably 2-3 mm, and the using amount ratio of the pear fruit sample to the ceramic beads is preferably 0.5-1.0 g: 3-5 granules.
In the invention, the rotation speed of the grinding is preferably 6-8 r.s-1The time is preferably 45-60 s. In the present invention, the grinding is preferably performed in a grinding pipe. In the embodiment of the present invention, after the grinding is completed, the grinding sample tube is preferably rinsed with ultrapure water, and the whole rinsed liquid is transferred to a centrifuge tube for leaching.
In the invention, the leaching is preferably carried out under ultrasonic conditions, and the leaching time is preferably 15-20 min. In the present invention, the ultrasonic conditions enable complete leaching.
In the invention, the heating temperature is preferably 80-90 ℃, the heating heat preservation time is preferably 30-35 min, and the heating function is to fully dissolve the sample in the extracting solution. In the invention, the heating temperature is preferably reached by adopting a water bath heating mode, and the water bath heating can reduce sample loss caused by liquid transfer and increase the real reliability of the experiment. In the invention, the sample container is preferably sealed in the heating process, so that the cover of the sample container is prevented from being cracked by heating in the water bath process. In the invention, the sample container is preferably shaken every 5-10 minutes in the heating process, so that the temperature of each part in the sample container is kept consistent.
In the invention, after naturally cooling to room temperature, the method preferably further comprises centrifuging the obtained cooling system to obtain a supernatant; and filtering the supernatant by using a 0.22 mu m water system filter membrane, wherein the filtrate is the liquid to be detected.
In the invention, the centrifugal temperature is preferably 4 ℃, and the rotating speed is preferably 12000-14000 r.min-1The time is preferably 15-20 min.
In the present invention, the water-based filter is preferably a 0.22 μm common water-based filter available from Shanghai Anpu (ANPEL).
After a solution to be detected is obtained, the solution to be detected is subjected to a first ultra high performance liquid chromatography detection to obtain the type of soluble sugar in a pear fruit sample, wherein the conditions of the first ultra high performance liquid chromatography detection comprise:
the high-efficiency liquid phase conditions include: a chromatographic column: UPLCACQUITYBEHAmide; mobile phase: the acetonitrile-ammonia water mixed solution and water are mixed according to the volume ratio of 85:15, the mass fraction of ammonia water in the acetonitrile-ammonia water mixed solution is 1%, and the flow rate is as follows: 0.2 mL/min-1Column temperature: 45 ℃, sample injection time: 15min, sample injection volume: 2 mu L of the solution;
the detection conditions include: ELSD detector, gas pressure: 25Psi, drift tube temperature: 55 ℃, sprayer: at 25 ℃.
In the present invention, the specification of the UPLCACQUITYBEHAMIde column is preferably 2.1mm × 100mm × 1.7 μm.
After the liquid to be detected is obtained, the liquid to be detected is subjected to second ultra high performance liquid chromatography detection to obtain the type of the organic acid in the pear fruit sample, wherein the conditions of the second ultra high performance liquid chromatography detection comprise:
the high-efficiency liquid phase conditions include: a chromatographic column: ACQUITYUPLCHSST 3; mobile phase: potassium dihydrogen phosphate solution-methanol, wherein the concentration of potassium dihydrogen phosphate in the mobile phase is 20mmol/L, the mass fraction of methanol is 1%, the pH value of the mobile phase is 2.4, and the flow rate is as follows: 0.25 mL/min-1Column temperature: 30 ℃, sample injection time: 5min, sample injection volume: 2 mu L of the solution;
the detection conditions include: PDA detector, wavelength: 214 nm.
In the present invention, the specification of the ACQUITYUPLCHSST3 column is preferably 100mm × 2.1mm × 1.8 μm.
The pH value of the mobile phase is preferably adjusted to 2.4 by using phosphoric acid, the concentration and the using amount of the phosphoric acid are not particularly limited, and the pH value of the mobile phase can be adjusted to 2.4.
In order to further illustrate the present invention, the following examples are provided to describe the qualitative detection method of sugar acid in pear fruit in detail, but they should not be construed as limiting the scope of the present invention.
Example 1
Preparing a soluble sugar mixed standard (comprising fructose, sorbitol, glucose and sucrose), wherein the ratio of fructose in the standard: glucose: sorbitol: the mass ratio of sucrose was 5:3:3:1, the concentration gradient of the mixed standard was 0.2, 0.4, 0.6, 0.8, 1.0mg/mL, and the mixture was measured and analyzed by ultra high performance liquid chromatography (Waters, H-class), and the liquid phase conditions were: mobile phase: acetonitrile (containing 1% by mass of ammonia water): the volume ratio of water is 85:15, flow rate: 0.2 mL/min-1Column temperature: 45 ℃, sample injection time: 15min, sample injection volume: 2 mu L of the solution; a detector: ELSD detector, gas 25Psi, drift tube temperature: 55 ℃, sprayer: 25 ℃; a chromatographic column: UPLCACQUITYBEHAMIde (2.1mm × 100mm × 1.7 μm), and the peak appearance of the obtained soluble sugar mixed standard is shown in FIG. 1, wherein 1 is fructose, and the peak appearance time is 4.751 min; 2 is sorbitol, the peak-out time is 5.953 min; 3 is glucose, the peak time is 6.452 min; 4 is sucrose, the peak-off time is 12.679min, and all components can completely peak within 14 min.
Preparing an organic acid mixed standard (including oxalic acid, quinic acid, malic acid, shikimic acid and citric acid), wherein the ratio of oxalic acid in the mixed standard is as follows: malic acid: shikimic acid: quinic acid: citric acid: the mass ratio of succinic acid is 0.2:1:0.02:2:1:1, the concentration gradient of the mixed standard is 0.2, 0.4, 0.6, 0.8 and 1.0mg/mL, the detection and analysis are carried out by using an ultra high performance liquid chromatograph (UPLCultimate3000), and the liquid phase conditions are as follows: mobile phase: 100% 20mmol/L potassium dihydrogen phosphate (containing 1% by mass of methanol), pH 2.4 (adjusted with phosphoric acid), flow rate: 0.25 mL/min-1Column temperature: 30 ℃, sample injection time: 5min, sample injection volume: 2 mu L of the solution; a detector: PDA detector, wavelength: 214 nm; a chromatographic column: ACQUITYUPLCHHSST 3(100mm × 2.1mm × 1.8 μm), wherein 1 is oxalic acid, and the peak emergence time is 1.030 min; 2 is quinic acid, and the peak-producing time is 1.273 min; 3 is malic acid, the peak-off time is 1.603 min; 4 is shikimic acid, the peak-off time is 1.743 min; 5 is citric acid, the peak-out time is 3.043 min; 6 is succinic acid, the peak-out time is 3.693min, and all components can completely peak out within 5 min.
Example 2
(1) The extraction process of the pear sugar acid comprises the following steps:
a. weighing 0.5 g-1.0 g of white pear fruit sample, grinding the white pear fruit sample into powder under the condition of liquid nitrogen, recording the weight of the sample, and repeating the steps for three samples.
b. Putting a weighed powdery pear fruit sample (0.5 g-1.0 g) into a 5mL sample grinding tube, adding 2mL ultrapure water and 3 ceramic beads (the diameter is 2mm), and grinding in a sample grinding machine (the rotating speed is 6m s)-145s) grinding to homogenate.
c. Transferring the sample in the step b into a 10mL centrifuge tube, rinsing the grinding tube with 3mL ultrapure water, and transferring all the rinsed liquid into the 10mL centrifuge tube.
d. And c, placing the 10mL centrifuge tube in the step c in an ultrasonic pot for ultrasonic treatment for 15min to fully leach.
e. And d, sealing the 10mL of centrifugal tube in the step d with a sealing film to prevent the cover from being cracked by heating in the water bath process, then placing the centrifugal tube in a water bath kettle at the temperature of 80 ℃ for 30min, and shaking the centrifugal tube every 5min to keep the temperature of each part in the tube consistent.
f. After the water-bathed sample was cooled to room temperature, it was centrifuged (4 ℃ C., 12000 r.min)-1) Taking supernatant after 15min, filtering the supernatant with 0.22 μm water system filter membrane into small brown bottle, and waiting for detection.
(2) The method for measuring the soluble sugar in the pear fruits comprises the following steps:
analyzing the sample obtained in the step (1) by using an ultra-high performance liquid chromatograph (Waters, H-class), wherein the liquid phase conditions are as follows: mobile phase: acetonitrile (containing 1% by massAmmonia): the volume ratio of water is 85:15, flow rate: 0.2 mL/min-1Column temperature: 45 ℃, sample injection time: 15min, sample injection volume: 2 mu L of the solution; a detector: ELSD detector, gas 25Psi, drift tube temperature: 55 ℃, sprayer: 25 ℃; a chromatographic column: UPLCACQUITYBEHAMIde (2.1 mm. times.100 mm. times.1.7 μm).
The peak appearance spectrogram of soluble sugar in fructus Pyri is shown in FIG 3, wherein 1 is fructose, and the peak appearance time is 4.703 min; 2 is sorbitol, the peak-out time is 5.768 min; 3 is glucose, the peak time is 6.354 min; 4 is sucrose, the peak-off time is 12.257min, and all components can completely peak within 14 min.
(3) The method for measuring the organic acid in the pear fruit comprises the following steps:
detecting and analyzing the sample extracted in the step (1) by using an ultra high performance liquid chromatograph (UPLCultimate3000), wherein the liquid phase conditions are as follows: mobile phase: 100% 20mmol/L potassium dihydrogen phosphate (containing 1% by mass of methanol), pH 2.4 (adjusted with phosphoric acid), flow rate: 0.25 mL/min-1Column temperature: 30 ℃, sample injection time: 5min, sample injection volume: 2 mu L of the solution; a detector: PDA detector, wavelength: 214 nm; a chromatographic column: ACQUITYUPLCHSST3(100 mm. times.2.1 mm. times.1.8 μm).
The peak spectrum of organic acid in pear fruit is shown in FIG. 4, wherein 1 is oxalic acid, and the peak time is 1.097 min; 2 is quinic acid, and the peak time is 1.297 min; 3 is malic acid, the peak-off time is 1.633 min; 4 is shikimic acid, the peak-off time is 1.767 min; 5 is citric acid, and the peak-off time is 3.130 min; the succinic acid content in pear pulp is not detected, and all components can completely peak within 5 min.
According to the qualitative determination method disclosed by the invention, the qualitative determination of the soluble sugar and the organic acid is realized by accurately limiting the conditions of the ultra-high performance liquid chromatography detection, the determination of the soluble sugar can be controlled to be complete within 14min, the determination of the organic acid can be controlled to be complete within 5min, the automation degree is high, the repeatability is good, the method can be popularized to the research of the sugar acid content and the components of fruits in the future, and the large-batch detection of the sugar acid content in the pear fruits can be realized; compared with the conventional sugar acid extraction method (the time for using a single sample is 3.5 hours), the method saves more time, and the conventional ethanol solution is changed into water in the extraction process, so that the use of organic solvents is reduced, and the safety of experiments is improved.
The foregoing is merely a preferred embodiment of the invention and is not intended to limit the invention in any manner. It should be noted that, for those skilled in the art, without departing from the principle of the present invention, several improvements and modifications can be made, and these improvements and modifications should also be construed as the protection scope of the present invention.
Claims (10)
1. A qualitative detection method for soluble sugar acid in pear fruits, wherein the sugar comprises fructose, sorbitol, glucose and sucrose, the acid is an organic acid, and the organic acid comprises oxalic acid, quinic acid, malic acid, shikimic acid and citric acid, and the method is characterized by comprising the following steps:
mixing a pear fruit sample, water and a grinding medium, and then sequentially grinding, leaching, heating and naturally cooling to room temperature to obtain a liquid to be detected;
performing first ultra-high performance liquid chromatography detection on the solution to be detected to obtain the species of soluble sugar in the pear fruit sample, wherein the conditions of the first ultra-high performance liquid chromatography detection comprise:
the high-efficiency liquid phase conditions include: a chromatographic column: UPLC acid BEH Amide; mobile phase: the acetonitrile-ammonia water mixed solution and water are mixed according to the volume ratio of 85:15, the mass fraction of ammonia water in the acetonitrile-ammonia water mixed solution is 1%, and the flow rate is as follows: 0.2 mL/min-1Column temperature: 45 ℃, sample injection time: 15min, sample injection volume: 2 mu L of the solution;
the detection conditions include: ELSD detector, gas pressure: 25Psi, drift tube temperature: 55 ℃, sprayer: 25 ℃;
and carrying out second ultra high performance liquid chromatography detection on the liquid to be detected to obtain the species of the organic acid in the pear fruit sample, wherein the conditions of the second ultra high performance liquid chromatography detection comprise:
the high-efficiency liquid phase conditions include: a chromatographic column: ACQUITY UPLC HSS T3; mobile phase: potassium dihydrogen phosphate solution-methanol, wherein the concentration of potassium dihydrogen phosphate in the mobile phase is 20mmol/L, 1% methanol by mass, pH 2.4 of the mobile phase, flow rate: 0.25 mL/min-1Column temperature: 30 ℃, sample injection time: 5min, sample injection volume: 2 mu L of the solution;
the detection conditions include: PDA detector, wavelength: 214 nm.
2. The qualitative detection method according to claim 1, wherein the dosage ratio of the pear fruit sample to water is 0.5-1.0 g: 2-4 mL.
3. The qualitative detection method according to claim 1, wherein the grinding medium is ceramic beads, and the particle size of the ceramic beads is 2-3 mm.
4. The qualitative detection method according to claim 1 or 3, wherein the rotation speed of the grinding is 6-8 m-s-1The time is 45-60 s.
5. The qualitative detection method according to claim 1, wherein the leaching is performed under ultrasonic conditions, and the leaching time is 15-20 min.
6. The qualitative detection method according to claim 1, wherein the heating temperature is 80 to 90 ℃ and the heating holding time is 30 to 35 min.
7. The qualitative detection method according to claim 1, wherein the step of naturally cooling to room temperature further comprises: centrifuging the obtained cooling system to obtain a supernatant; and filtering the supernatant by using a 0.22 mu m water system filter membrane, wherein the filtrate is the liquid to be detected.
8. The qualitative detection method according to claim 7, wherein the centrifugation temperature is 4 ℃ and the rotation speed is 12000-14000 r-min-1The time is 15-20 min.
9. The qualitative detection method according to claim 1, wherein the specification of the UPLC acid BEH Amide chromatography column is 2.1mm x 100mm x 1.7 μm.
10. The qualitative detection method according to claim 1, wherein the ACQUITY UPLC HSS T3 chromatographic column has a specification of 100mm x 2.1mm x 1.8 μm.
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