CN107907616B - Method for detecting erythromycin residue in mushroom dregs - Google Patents
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Abstract
A method for detecting erythromycin residue in mushroom dregs relates to a method for detecting the titer of the erythromycin residue in mushroom dregs, and comprises the steps of preparing a reference substance solution and a test substance solution, then injecting the standard substance solution and the test substance solution into a liquid chromatograph by adopting a high performance liquid chromatography in a gradient elution mode, and calculating the titer of the erythromycin according to an external standard method. The invention establishes a liquid chromatography analysis method of erythromycin in the mushroom dregs through optimization of chromatographic conditions and tests of preparation methods of a test sample and a standard substance; a method for detecting erythromycin in mushroom dregs is established by optimizing a chromatographic separation system, and proved by methodological verification test research: the established method has high accuracy, strong specificity and good reproducibility, and can effectively detect the titer of the erythromycin in the mushroom dregs.
Description
Technical Field
The invention relates to the technical field of detection and inspection of mushroom dregs, in particular to a method for detecting the residual titer of erythromycin in mushroom dregs.
Background
Erythromycin (Erythromycin) was discovered by Mc Guire et al in 1952 as the main species of the tetradecanoid macrolide antibiotics produced by Streptomyces erythraeus (Streptomyces erythraeus). Erythromycin is classified into erythromycin A, B, C and erythromycin D, wherein erythromycin A has antibacterial activity and is the main active ingredient of erythromycin. Although the antibacterial spectrum and antibacterial activity of erythromycin B are similar to those of erythromycin A, the toxicity of erythromycin B is relatively high. Erythromycin C is not an active substance and is a main impurity of erythromycin produced in China. Pharmaceutical workers in various countries in the world have conducted a series of extensive and intensive researches on chemical structures and antibacterial activities of erythromycin according to the principle of erythromycin acid ketal formation failure, and a large number of erythromycin macrolides with new pharmacokinetic properties are obtained: derivatives such as roxithromycin, azithromycin, clarithromycin, dirithromycin and fluoroerythromycin. The advent of these derivatives has further expanded the market share that macrolide antibiotics have in anti-infective drugs.
The erythromycin bacterium residue is waste residue produced after erythromycin is produced by adding erythromycin bacterium, namely saccharopolyspora erythraea, into starch, soybean meal and corn steep liquor which are used as main raw materials and aerobic submerged fermentation, and the main components of the erythromycin bacterium residue are residual starch, other saccharides, residual soybean meal, a large amount of mycelia, partial inorganic matters and a small amount of erythromycin residue (the concentration in a bacterium residue stock solution is about 600-1400U/mL).
The erythromycin mushroom dregs emit a special peculiar smell, the content of organic matters in the mushroom dregs is high, secondary fermentation can be caused if the mushroom dregs are discarded at will without treatment, the color becomes black, foul smell is generated, a large amount of flies are attracted, and the environment is very bad. According to the revised national records of dangerous waste of 2008, antibiotic fungi residues belong to the waste of culture medium in the production process of chemical bulk drugs and need to be managed according to the dangerous waste. The antibiotic residues contain residual antibiotics, metabolic intermediates and the like, are special dangerous wastes, can generate potential harm to ecological environment and human health if being improperly disposed, and have the characteristics of concealment, hysteresis, accumulation, cooperativity, linkage and the like.
Disclosure of Invention
Aiming at the current situation that a standard detection method for erythromycin residue in mushroom dregs is not available at home and abroad at present, the invention provides a method for detecting erythromycin residue in mushroom dregs, which can effectively solve the influence of matrix effect and solve the problems that the prior art can not quickly/accurately detect the erythromycin residue in mushroom dregs.
The invention discloses a method for detecting erythromycin residue in mushroom dregs, which is carried out according to the following steps:
firstly, preparing a sample to be tested:
1) weighing bacteria residue containing erythromycin, adding extracting agent acetonitrile-Tris-CaCl2Performing vortex oscillation on the solution for 45-60 s, performing auxiliary extraction in ultrasound for 20-30 min, centrifuging at 3500-4000 rpm for 5-10 min, taking supernatant, and collecting precipitate;
2) adding an extracting agent acetonitrile-Tris-CaCl into the precipitate collected in the step 1)2Performing vortex oscillation on the solution for 45-60 s, performing auxiliary extraction in ultrasound for 20-30 min, centrifuging at 3500-4000 rpm for 5-10 min, and taking supernatant;
3) mixing the supernatants obtained in the step 1) and the step 2), placing the mixed solution in a water bath at 35 ℃, and performing rotary evaporation to obtain an inorganic phase, wherein the volume of the supernatant is one fifth of that of the supernatant;
4) adding Tris-CaCl with the pH value of 4-7 into the inorganic phase2Performing oscillation and vortex on the solution again, adjusting the pH value to 8-10 after the residues are completely dissolved, adding a dispersing agent methanol and an extracting agent 1, 2-dichloroethane, performing vortex for 30s, centrifuging for 5min at 4000r/min, and extracting the lower-layer liquid;
5) adding an extracting agent 1, 2-dichloroethane into the lower layer liquid in the step 4), performing vortex for 30-45 s, centrifuging at 4000r/min for 5min, and extracting the lower layer liquid;
6) repeating the operations of the steps 1) to 5), and extracting the lower-layer liquid;
7) combining the lower layer liquid obtained in the step 5) and the lower layer liquid obtained in the step 6), blowing the lower layer liquid to be nearly dry by using nitrogen, redissolving the lower layer liquid by using methanol, and filtering the redissolved liquid by using a 0.45-micrometer filter membrane to obtain a sample of a test sample;
wherein the erythromycin bacterium residue and an extracting agent acetonitrile-Tris-CaCl2The mass-to-volume ratio of the solution is 1 g: 12-17 mL; precipitating and extracting agent acetonitrile-Tris-CaCl2The mass-to-volume ratio of the solution is 1 g: 12-17 mL; inorganic phase and Tris-CaCl2The volume ratio of the complex solution is 4-8: 10; the volume ratio of the inorganic phase to the dispersant methanol is 4-8: 1; the volume ratio of the inorganic phase to the extractant 1, 2-dichloroethane is 4-8: 1;
secondly, drawing a standard curve:
diluting an erythromycin standard stock solution into standard working solutions with mass concentrations of 1.00mg/L, 5.00mg/L, 10.00mg/L, 20.00mg/L, 50.00mg/L, 100.00mg/L, 200.00mg/L, 500.00mg/L and 1000.00mg/L by using methanol, detecting the standard working solutions by using a high performance liquid chromatography, drawing a standard curve by using the measured peak area as a vertical coordinate and the concentration of the standard working solutions as a horizontal coordinate, and solving a regression equation and a correlation coefficient;
thirdly, determination: sucking the standard working solution with the concentration of 500.00mg/L and the sample to be tested, respectively injecting the standard working solution and the sample to be tested into a liquid chromatograph, and calculating the residual titer of the erythromycin according to an external standard method; thus completing the detection method of the erythromycin residue in the mushroom dregs;
chromatographic conditions are as follows: a chromatographic column: the chromatographic column is a Waters-C18 column, the inner diameter of the chromatographic column is 4.6mm, the length of the chromatographic column is 250mm, the diameter of the filler particles is 5 μm, and the ratio of the mobile phase: acetonitrile-0.01 moL/L dipotassium hydrogen phosphate solution system isocratic elution, detection wavelength: 215nm, flow rate: 1mL/min, column temperature: 20 to 35 ℃.
The invention has the following beneficial effects:
the invention establishes a high-efficiency liquid phase-ultraviolet detector detection analysis method of erythromycin in erythromycin mushroom residue through the experiments of the chromatographic optimization and the preparation methods of a test sample and a standard substance; a method for detecting erythromycin in the mushroom dregs is established by optimizing chromatographic conditions, and proved by methodological verification test research: the established method has high accuracy, strong specificity and good reproducibility, and can effectively detect the residual quantity of the erythromycin in the mushroom dregs. By verification, the absolute difference between two independent measurements obtained under repetitive conditions for the protocol of the invention must not exceed 10% of the arithmetic mean. The addition level was 200mg/kg and the process recovery was 96.32%. The method has a qualitative detection limit of 0.0660mg/Kg and a quantitative detection limit of 0.2210 mg/Kg. The RSD was 8.85%.
The method can effectively extract the erythromycin from the mushroom dregs, and reduces the influence of other impurities in the mushroom dregs on subsequent detection as much as possible, thereby improving the accuracy of the detection.
Drawings
FIG. 1 is a high performance liquid chromatogram of an erythromycin standard sample from example 1;
FIG. 2 is a high performance liquid chromatogram of erythromycin content in the mushroom dregs of example 1;
FIG. 3 is a high performance liquid chromatogram of an erythromycin standard sample of example 2;
FIG. 4 is a high performance liquid chromatogram of erythromycin content in the mushroom dregs of example 2.
Detailed Description
The first embodiment is as follows: the method for detecting the erythromycin residue in the mushroom dregs in the embodiment is carried out according to the following steps:
firstly, preparing a sample to be tested:
1) weighing bacteria residue containing erythromycin, adding extracting agent acetonitrile-Tris-CaCl2Performing vortex oscillation on the solution for 45-60 s, performing auxiliary extraction in ultrasound for 20-30 min, centrifuging at 3500-4000 rpm for 5-10 min, taking supernatant, and collecting precipitate;
2) adding an extracting agent acetonitrile-Tris-CaCl into the precipitate collected in the step 1)2Performing vortex oscillation on the solution for 45-60 s, performing auxiliary extraction in ultrasound for 20-30 min, centrifuging at 3500-4000 rpm for 5-10 min, and taking supernatant;
3) mixing the supernatants obtained in the step 1) and the step 2), placing the mixed solution in a water bath at 35 ℃, and performing rotary evaporation to obtain an inorganic phase, wherein the volume of the supernatant is one fifth of that of the supernatant;
4) adding Tris-CaCl with the pH value of 4-7 into the inorganic phase2Performing oscillation and vortex on the solution again, adjusting the pH value to 8-10 after the residues are completely dissolved, adding a dispersing agent methanol and an extracting agent 1, 2-dichloroethane, performing vortex for 30s, centrifuging for 5min at 4000r/min, and extracting the lower-layer liquid;
5) adding an extracting agent 1, 2-dichloroethane into the lower layer liquid in the step 4), performing vortex for 30-45 s, centrifuging at 4000r/min for 5min, and extracting the lower layer liquid;
6) repeating the operations of the steps 1) to 5), and extracting the lower-layer liquid;
7) combining the lower layer liquid obtained in the step 5) and the lower layer liquid obtained in the step 6), blowing the lower layer liquid to be nearly dry by using nitrogen, redissolving the lower layer liquid by using methanol, and filtering the redissolved liquid by using a 0.45-micrometer filter membrane to obtain a sample of a test sample;
wherein the erythromycin bacterium residue and an extracting agent acetonitrile-Tris-CaCl2The mass-to-volume ratio of the solution is 1 g: 12-17 mL; precipitating and extracting agent acetonitrile-Tris-CaCl2The mass-to-volume ratio of the solution is 1 g: 12-17 mL; inorganic phase and Tris-CaCl2The volume ratio of the complex solution is 4-8: 10; the volume ratio of the inorganic phase to the dispersant methanol is 4-8: 1; the volume ratio of the inorganic phase to the extractant 1, 2-dichloroethane is 4-8: 1;
secondly, drawing a standard curve:
diluting an erythromycin standard stock solution into standard working solutions with mass concentrations of 1.00mg/L, 5.00mg/L, 10.00mg/L, 20.00mg/L, 50.00mg/L, 100.00mg/L, 200.00mg/L, 500.00mg/L and 1000.00mg/L by using methanol, detecting the standard working solutions by using a high performance liquid chromatography, drawing a standard curve by using the measured peak area as a vertical coordinate and the concentration of the standard working solutions as a horizontal coordinate, and solving a regression equation and a correlation coefficient;
thirdly, determination: sucking 10uL of each of the standard working solution with the concentration of 500.00mg/L and the sample to be tested, respectively injecting the standard working solution and the sample to be tested into a liquid chromatograph, and calculating the residual titer of the erythromycin according to an external standard method; thus completing the detection method of the erythromycin residue in the mushroom dregs;
chromatographic conditions are as follows: a chromatographic column: the chromatographic column is a Waters-C18 column, the inner diameter of the chromatographic column is 4.6mm, the length of the chromatographic column is 250mm, the diameter of the filler particles is 5 μm, and the ratio of the mobile phase: acetonitrile-0.01 moL/L dipotassium hydrogen phosphate solution system isocratic elution, detection wavelength: 215nm, flow rate: 1mL/min, column temperature: 20 to 35 ℃.
The second embodiment is as follows: the first difference between the present embodiment and the specific embodiment is: the pH of the dipotassium hydrogen phosphate solution is 8-10. The rest is the same as the first embodiment.
The third concrete implementation mode: the first difference between the present embodiment and the specific embodiment is: the residual quantity of the erythromycin in the mushroom dregs is 500-1000 mg/kg. The rest is the same as the first embodiment.
The fourth concrete implementation mode: the first difference between the present embodiment and the specific embodiment is: the residual quantity of the erythromycin in the mushroom dregs is 800-1000 mg/kg. The rest is the same as the first embodiment.
The fifth concrete implementation mode: the first difference between the present embodiment and the specific embodiment is: the method has a minimum qualitative detection limit of 0.0660mg/Kg and a quantitative detection limit of 0.2210 mg/Kg. The rest is the same as the first embodiment.
The sixth specific implementation mode: the first difference between the present embodiment and the specific embodiment is: the method has a minimum qualitative detection limit of 0.0660mg/Kg and a quantitative detection limit of 0.2210 mg/Kg. The rest is the same as the first embodiment.
The seventh embodiment: the first difference between the present embodiment and the specific embodiment is: precipitating and extracting agent acetonitrile-Tris-CaCl2The mass-to-volume ratio of the solution is 1 g: 15 mL. The rest is the same as the first embodiment.
The specific implementation mode is eight: the first difference between the present embodiment and the specific embodiment is: inorganic phase and Tris-CaCl2The volume ratio of the complex solution is 6: 10. The rest is the same as the first embodiment.
The specific implementation method nine: the first difference between the present embodiment and the specific embodiment is: the volume ratio of the inorganic phase to the dispersant methanol was 6: 1. The rest is the same as the first embodiment.
The detailed implementation mode is ten: the first difference between the present embodiment and the specific embodiment is: the volume ratio of the inorganic phase to the extractant 1, 2-dichloroethane was 6: 1. The rest is the same as the first embodiment.
The invention is not limited to the above embodiments, and one or a combination of several embodiments may also achieve the object of the invention.
The beneficial effects of the present invention are demonstrated by the following examples:
example 1
The method for detecting the residual titer of the erythromycin in the mushroom dregs comprises the following steps:
the instrument comprises the following steps: high performance liquid chromatograph
Chromatographic conditions are as follows: a chromatographic column: Waters-C18 column (4.6 mm. times.250 mm, 5 μm), mobile phase: acetonitrile-0.01 moL/L dipotassium hydrogen phosphate solution (pH 10) system isocratic elution, detection wavelength: 215nm, column temperature: 35 ℃ is carried out.
1) Preparing a sample to be tested: weighing 1g (accurate to 0.0001g) of sample, adding 15mL of extractant acetonitrile-Tris-CaCl into a 50mL polypropylene centrifuge tube2Vortex and shake the solution for 1min, extract for 30min under ultrasound, centrifuge at 3000rpm for 10min, and collect the supernatant. The operation is repeated to extract the supernatant once, and the two supernatants are combined. Performing rotary evaporation on the extract in 35 deg.C water bath until 6mL inorganic phase remains, transferring into 50mL polypropylene centrifuge tube, and adding Tris-CaCl with pH of 52And mixing 10mL of the solution, sufficiently shaking and vortexing for 1min, transferring the solution to a 50mL polypropylene centrifuge tube after the residues are completely dissolved, adding 400u L ammonia water to adjust the pH value to 10, and adding 1mL of dispersant methanol. Injecting 1mL of 1, 2-dichloroethane as an extracting agent rapidly, vortexing for 30s, centrifuging for 5min at 4000r/min, extracting the lower layer liquid, injecting 1mL of 1, 2-dichloroethane rapidly again, repeating the previous operation, combining the 1, 2-dichloroethane containing the target substance, blowing nitrogen to be nearly dry, and then redissolving with 1mL of methanol. The compound solution is filtered through a 0.45 mu m filter membrane to obtain the compound solution.
2) Drawing a standard product curve: and (3) diluting the standard stock solution into standard solutions with mass concentrations of 1.00mg/L, 5.00mg/L, 10.00mg/L, 20.00mg/L, 50.00mg/L, 100.00mg/L, 200.00mg/L, 500.00mg/L and 1000.00mg/L by using methanol, detecting the standard working solution by using high performance liquid chromatography, and drawing a standard curve by taking the peak area as an ordinate and the corresponding standard solution concentration as an abscissa. And solving a regression equation and a correlation coefficient.
3) And (3) determination: and (3) sucking the standard solution and the test solution 10u L, injecting into a liquid chromatograph, and calculating the erythromycin residual titer according to an external standard method.
According to the method for detecting the residual titer of the erythromycin in the mushroom dregs, the test process finds that: the impurities in the mushroom dregs are more, the existing method can not effectively detect the erythromycin, the detection amount is small, and the sensitivity is low. Quantitative control is required, and the difficulty is high; meanwhile, the sample treatment process is complex and tedious, so that part of erythromycin is lost. Through a large number of experiments, samples are extracted and purified according to the properties of various impurities, and meanwhile, a isocratic elution mode is adopted, so that trace erythromycin is effectively detected, efficient separation is achieved, and the titer of the erythromycin in the bacterial residues is effectively detected.
The verification result shows that: the average recovery for the accuracy test was: 96.32 percent. The RSD of the precision test is 8.85 percent; the linear relation is good, and the correlation coefficient gamma is 0.999; specificity tests investigate the absence of negative interference. Experiments prove that the established method has high accuracy, strong specificity and good reproducibility, and can effectively detect the residual titer of the erythromycin in the mushroom dregs.
Example 2
The method for detecting the erythromycin residue in the mushroom dregs comprises the following steps:
the instrument comprises the following steps: waters 2695 model high performance liquid chromatograph
Chromatographic conditions are as follows: a chromatographic column: Waters-C18 column (4.6 mm. times.250 mm, 5 μm), mobile phase: acetonitrile-0.01 moL/L dipotassium hydrogen phosphate solution (pH 10) system isocratic elution, detection wavelength: 215nm, flow rate: 1mL/min, column temperature: 35 ℃ is carried out.
1) Preparing a sample to be tested: weighing 1g (accurate to 0.0001g) of sample, adding 15mL of extractant acetonitrile-Tris-CaCl into a 50mL polypropylene centrifuge tube2Vortex the solution (7) for 1min, extracting with ultrasound for 20min, centrifuging at 3000rpm for 10min, and collecting supernatant. The procedure was repeated once and the supernatants were combined. Rotary evaporating the extractive solution in 35 deg.C water bath until 6ml inorganic phase remains, transferring into 50ml polypropylene centrifuge tube, and adding Tris-CaCl with pH of 62And (3) mixing 10mL of the solution, fully shaking and vortexing for 1min, transferring the solution to a 50mL polypropylene centrifuge tube after residues are completely dissolved, adding 400uL of ammonia water to adjust the pH value to 9, and adding 1mL of dispersant methanol. Injecting 1mL of 1, 2-dichloroethane as an extracting agent rapidly, vortexing for 30s, centrifuging for 5min at 4000r/min, extracting the lower layer liquid, injecting 1mL of 1, 2-dichloroethane rapidly again, repeating the previous operation, combining the 1, 2-dichloroethane containing the target substance, blowing nitrogen to be nearly dry, and then redissolving with 1mL of methanol. The compound solution is filtered through a 0.45 mu m filter membrane to obtain the compound solution.
2) Drawing a standard product curve: diluting the standard stock solution with methanol to standard solutions with mass concentrations of 1.00mg/L, 5.00mg/L, 10.00mg/L, 20.00mg/L, 50.00mg/L, 100.00mg/L, 200.00mg/L, 500.00mg/L and 1000.00mg/L, detecting the standard working solution by high performance liquid chromatography, drawing a standard curve by taking the measured peak area as a vertical coordinate and the corresponding standard solution concentration as a horizontal coordinate, and solving a regression equation and a correlation coefficient.
3) And (3) determination: and (3) sucking 10uL of the standard substance solution and the test sample solution, injecting into a liquid chromatograph, and calculating the erythromycin residual titer according to an external standard method.
The content of erythromycin A in the sample was calculated by the formula (1).
In the formula: x-the amount of erythromycin A in the sample in micrograms per gram (μ g/g);
c-concentration of tylosin in the sample solution in micrograms per milliliter (. mu.g/mL);
m is the sample mass in grams (g);
v-volume of sample solution in milliliters (mL).
The absolute difference between two independent measurements obtained under repetitive conditions must not exceed 10% of the arithmetic mean. The addition level was 200mg/kg and the process recovery was 96.32%. The method has a qualitative detection limit of 0.0660mg/Kg and a quantitative detection limit of 0.2210 mg/Kg. The RSD was 8.85%.
Claims (9)
1. A method for detecting erythromycin residue in mushroom dregs is characterized by comprising the following steps:
firstly, preparing a sample to be tested:
1) weighing bacteria residue containing erythromycin, adding extracting agent acetonitrile-Tris-CaCl2Performing vortex oscillation on the solution for 45-60 s, performing auxiliary extraction in ultrasound for 20-30 min, centrifuging at 3500-4000 rpm for 5-10 min, taking supernatant, and collecting precipitate;
2) adding an extracting agent acetonitrile-Tris-CaCl into the precipitate collected in the step 1)2Performing vortex oscillation on the solution for 45-60 s, performing auxiliary extraction in ultrasound for 20-30 min, centrifuging at 3500-4000 rpm for 5-10 min, and taking supernatant;
3) mixing the supernatants obtained in the step 1) and the step 2), placing the mixed solution in a water bath at 35 ℃, and performing rotary evaporation to obtain an inorganic phase, wherein the volume of the supernatant is one fifth of that of the supernatant;
4) adding Tris-CaCl with the pH value of 4-7 into the inorganic phase2Performing oscillation and vortex on the solution again, adjusting the pH value to 8-10 after the residues are completely dissolved, adding a dispersing agent methanol and an extracting agent 1, 2-dichloroethane, performing vortex for 30s, centrifuging for 5min at 4000r/min, and extracting the lower-layer liquid;
5) adding an extracting agent 1, 2-dichloroethane into the lower layer liquid in the step 4), performing vortex for 30-45 s, centrifuging at 4000r/min for 5min, and extracting the lower layer liquid;
6) repeating the operations of the steps 1) to 5), and extracting the lower-layer liquid;
7) combining the lower layer liquid obtained in the step 5) and the lower layer liquid obtained in the step 6), blowing the lower layer liquid to be nearly dry by using nitrogen, redissolving the lower layer liquid by using methanol, and filtering the redissolved liquid by using a 0.45-micrometer filter membrane to obtain a sample of a test sample;
wherein the erythromycin bacterium residue and an extracting agent acetonitrile-Tris-CaCl2The mass-to-volume ratio of the solution is 1 g: 12-17 mL; precipitating and extracting agent acetonitrile-Tris-CaCl2The mass-to-volume ratio of the solution is 1 g: 12-17 mL; inorganic phase and Tris-CaCl2The volume ratio of the complex solution is 4-8: 10; the volume ratio of the inorganic phase to the dispersant methanol is 4-8: 1; the volume ratio of the inorganic phase to the extractant 1, 2-dichloroethane is 4-8: 1;
secondly, drawing a standard curve:
diluting an erythromycin standard stock solution into standard working solutions with mass concentrations of 1.00mg/L, 5.00mg/L, 10.00mg/L, 20.00mg/L, 50.00mg/L, 100.00mg/L, 200.00mg/L, 500.00mg/L and 1000.00mg/L by using methanol, detecting the standard working solutions by using a high performance liquid chromatography, drawing a standard curve by using the measured peak area as a vertical coordinate and the concentration of the standard working solutions as a horizontal coordinate, and solving a regression equation and a correlation coefficient;
thirdly, determination: sucking the standard working solution with the concentration of 500.00mg/L and the sample to be tested, respectively injecting the standard working solution and the sample to be tested into a liquid chromatograph, and calculating the residual titer of the erythromycin according to an external standard method; thus completing the detection method of the erythromycin residue in the mushroom dregs;
chromatographic conditions are as follows: a chromatographic column: the chromatographic column is a Waters-C18 column, the inner diameter of the chromatographic column is 4.6mm, the length of the chromatographic column is 250mm, the diameter of the filler particles is 5 μm, and the ratio of the mobile phase: acetonitrile-0.01 moL/L dipotassium hydrogen phosphate solution system isocratic elution, detection wavelength: 215nm, flow rate: 1mL/min, column temperature: 20-35 ℃; the residual quantity of the erythromycin in the mushroom dregs is 500-1000 mg/kg.
2. The method for detecting erythromycin residue in mushroom dregs according to claim 1, wherein the pH of the dipotassium hydrogen phosphate solution is 8-10.
3. The method for detecting erythromycin residue in mushroom dregs of claim 1, wherein the method has a minimum qualitative detection limit of 0.0660mg/Kg and a quantitative detection limit of 0.2210 mg/Kg.
4. The method for detecting erythromycin residue in mushroom dregs according to claim 1, wherein the pH value adjustment in the step 4) is performed by using ammonia water.
5. The method of claim 1, wherein the erythromycin residue is extracted from the erythromycin residue with an extractant acetonitrile-Tris-CaCl2The mass-to-volume ratio of the solution is 1 g: 15 mL.
6. The method of claim 1, wherein the precipitate and the extractant acetonitrile-Tris-CaCl are used for detecting erythromycin residue in the mushroom dregs2The mass-to-volume ratio of the solution is 1 g: 15 mL.
7. The method for detecting erythromycin residue in mushroom dregs of claim 1, wherein the inorganic phase is Tris-CaCl2The volume ratio of the complex solution is 6: 10.
8. The method for detecting the erythromycin residue in the mushroom dregs according to claim 1, wherein the volume ratio of the inorganic phase to the dispersant methanol is 6: 1.
9. The method for detecting the erythromycin residue in the mushroom dregs according to claim 1, wherein the volume ratio of the inorganic phase to the 1, 2-dichloroethane as the extractant is 6: 1.
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