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CN-122017092-A - Method for detecting residue of Hainanmycin in poultry meat, eggs and egg products and pretreatment method

CN122017092ACN 122017092 ACN122017092 ACN 122017092ACN-122017092-A

Abstract

The invention provides a method for detecting residue of Hainanmycin in poultry meat, eggs and egg products and a pretreatment method, belonging to the technical field of food detection. The pretreatment method comprises the steps of (1) adding an extraction solvent into a sample to be detected for extraction, (2) adding anhydrous magnesium sulfate and sodium chloride, (3) centrifuging to obtain a supernatant, (4) loading the supernatant onto a solid-phase extraction column for purification and collection to obtain a first effluent, (5) loading the extraction solvent onto the solid-phase extraction column for collection to obtain a second effluent, and combining all the effluents, and (6) concentrating, redissolving and filtering all the effluents to obtain a sample liquid to be detected. The invention also discloses a method for detecting the residue of the Hainanmycin by liquid-mass spectrometry after the pretreatment of the sample, which has the characteristics of rapidness, high efficiency, low detection limit and good reproducibility, fills the blank of the method for detecting the Hainanmycin in animal-derived foods, and is suitable for trace detection of the Hainanmycin.

Inventors

  • QI HEMING
  • CUI FENGYUN
  • ZHANG PAN
  • WANG QI
  • LIU NA
  • LI ZIYI
  • ZHAO DAN
  • WANG PEIYUE
  • ZHANG CHAOHUI

Assignees

  • 中国海关科学技术研究中心

Dates

Publication Date
20260512
Application Date
20260227

Claims (10)

  1. 1. A pretreatment method for detecting the residue of Hainanmycin in poultry meat, eggs and egg products is characterized by comprising the following steps: (1) Acetonitrile is added into a sample to be detected for extraction, wherein the sample to be detected is poultry muscle, poultry liver, fresh eggs, liquid egg products, frozen egg products, processed eggs or dried egg products; (2) Adding anhydrous magnesium sulfate and sodium chloride into the mixed system obtained in the step (1), oscillating, centrifuging, and separating to obtain a supernatant; (3) Loading the supernatant obtained in the step (2) onto an acetonitrile-activated neutral alumina solid phase extraction column, and collecting a first effluent; (4) After the loading of the step (3) is completed, continuously washing the neutral alumina solid phase extraction column with acetonitrile, collecting a second effluent and combining the second effluent with the first effluent of the step (3); (5) And (3) concentrating all the liquid obtained after the step (4) is combined, redissolving the liquid in acetonitrile, and filtering to obtain the liquid to be tested.
  2. 2. The pretreatment method for detecting the residue of Hainanmycin in poultry meat, eggs and egg products according to claim 1, wherein in the step (1), when the sample to be detected is poultry muscle, poultry liver, fresh eggs, liquid egg products or ice egg products, the ratio of the sample to be detected to acetonitrile is 5g to (15-25) mL, when the sample to be detected is a reconstituted egg or dry egg products, water is added into the sample to be detected for homogenization, then acetonitrile is added for extraction, wherein the ratio of the reconstituted egg to water is 5g to (8-12) mL, the ratio of the reconstituted egg to acetonitrile is 5g to (15-25) mL, the ratio of the dry egg products to water is 1g to (2.5-3.5) mL, and the ratio of the dry egg products to acetonitrile is 1g to (15-25) mL.
  3. 3. The pretreatment method for detecting a residue of Hainanmycin in poultry meat, eggs and egg products according to claim 1, wherein the mass ratio of the poultry muscle, poultry liver, fresh egg, liquid egg product, frozen egg product or processed egg sample to anhydrous magnesium sulfate, sodium chloride is 5:3:0.5; the mass ratio of the dry egg product sample to anhydrous magnesium sulfate to sodium chloride is 1:3:0.5.
  4. 4. The pretreatment method for detecting a residue of hainanmycin in poultry, eggs and egg products as claimed in claim 1, wherein in the step (3), the mass of the filler of the neutral alumina solid phase extraction column is 500mg; During sample loading, the mass ratio of the sample amount in the supernatant obtained by poultry muscle, poultry liver, fresh eggs, liquid egg products, ice egg products or processed egg samples to the neutral alumina solid phase extraction column filler is (1.75-2.25) to 1, and the mass ratio of the sample amount in the supernatant obtained by dry egg products to the neutral alumina solid phase extraction column filler is (0.35-0.45) to 1.
  5. 5. The pretreatment method for detecting a residue of Hainanmycin in poultry, eggs and egg products as claimed in claim 4, wherein an anhydrous sodium sulfate layer with a height of 0.5cm is pre-packed on the filler upper layer of said neutral alumina solid phase extraction column.
  6. 6. The pretreatment method for detecting the residue of Hainanmycin in poultry, eggs and egg products as claimed in claim 1, wherein in the step (5), the concentration is carried out by adopting a nitrogen blow-drying mode, the temperature is lower than 40 ℃, and the pore diameter of the filtering membrane is 0.22 μm.
  7. 7. A method for detecting a residue of hainanmycin in poultry meat, eggs and egg products, comprising the steps of: The pretreatment method of any one of claims 1 to 6 is used for treating a sample to obtain a liquid to be tested, and the high performance liquid chromatography-tandem mass spectrometry is used for detecting the Hainanmycin in the liquid to be tested.
  8. 8. The method according to claim 7, wherein the detection conditions of the high performance liquid chromatography are: ACQUITY UPLC BEH C18, 2.1X100 mm,1.7 μm; Column temperature is 30 ℃; Mobile phase A is 5mmol/L ammonium acetate aqueous solution containing 0.1wt% formic acid, mobile phase B is acetonitrile containing 0.1wt% formic acid; the flow rate is 0.5mL/min; Sample injection amount is 5 mu L; The elution mode is gradient elution.
  9. 9. The method according to claim 8, wherein the gradient elution is performed in an elution program of 0-0.5min,35% A, 0.5-4.0min,35% A-2% A, 4.0-10.0min,2% A, 10.1-13.0min,35% A.
  10. 10. The method of claim 7, wherein the detection conditions of tandem mass spectrometry are: a mass analyzer, triple quadrupole rods; ion source ESI+; The ion source spray voltage is 5500V; ion source temperature 400 ℃; the air pressure of the air curtain is 30Psi; The atomization air pressure is 35Psi; Heating auxiliary air pressure to 35Psi; The collision cell emits a voltage of 10V.

Description

Method for detecting residue of Hainanmycin in poultry meat, eggs and egg products and pretreatment method Technical Field The invention belongs to the technical field of food detection, and particularly relates to a method for detecting the residue of Hainanmycin in poultry meat, eggs and egg products and a pretreatment method. Background Hainanmycin (HAINANMYCIN) is a polyether ionophore anticoccidial which is independently developed, and plays an important role in preventing and treating chicken coccidiosis after being automatically put into use. However, toxicology studies indicate that the residue of Hainanmycin in animal food may constitute a potential risk to consumer health. In view of the importance, a sensitive, accurate and reliable detection method is established, is used for monitoring the residual level of the Hainanmycin in poultry meat, poultry eggs and egg products, and has urgent practical significance for guaranteeing food safety. At present, the research on a detection method of the residue of the Hainanmycin in animal foods is not perfect, and a standard system is blank. The detection means in the prior art mainly has the following problems that (1) the matrix interference is serious, the purification effect is poor, the matrixes of poultry meat (especially liver), poultry eggs and egg products (such as yolk powder and preserved eggs) are extremely complex, and the matrixes are rich in fat, protein, phospholipid, endogenous pigments (such as carotenoid in yolk) and the like. These substances can produce serious ion suppression or enhancement effects (matrix effects) in mass spectrometry detection and contaminate the chromatographic column and ion source. The existing pretreatment method, such as simple liquid-liquid extraction (for example, adopting acetonitrile saturated n-hexane to remove lipid), has a certain effect on removing protein and partial lipid, but has limited removal capability on components such as small molecular polar interferents, pigments and the like, so that the background interference of the final sample liquid is high, and the detection sensitivity and accuracy are difficult to ensure. (2) The pretreatment flow is complicated or the recovery rate is unstable, and some solid phase extraction methods which are used for detection of other veterinary drugs, such as a hydrophilic-lipophilic balance (HLB) and other general solid phase extraction columns, often require complex activation, leaching and elution steps to balance the recovery rate and the purification effect when facing the polyether compound with the specific structure of the Hainanmycin. The process is complicated, the time consumption is long, and the recovery rate of the Hainanmycin in a strong hydrophilic or strong ion exchange environment is low or has large fluctuation due to the non-ideal retention behavior. In addition, the traditional solid phase extraction method has the problems of easy blockage of small columns, limited sample loading and the like. (3) There is a lack of systematic approach to avian egg products, a few literature approaches to Hainanmycin have focused on feed or animal tissue, and there is a lack of systematic pretreatment studies on egg products of varying morphology and composition (e.g., liquid eggs, salted processed eggs, dried processed egg powder). In particular to a method for ensuring stable extraction and efficient purification of the Hainanmycin for processed egg products (such as preserved eggs and salted eggs) with salt content and pH value changed. In addition, the purification methods related to the Hainanmycin in the prior art mostly adopt liquid-liquid extraction or general solid phase extraction columns (such as HLB), but the methods have limited purification effects when facing high-fat and high-pigment poultry meat and egg products, and the Hainanmycin has unstable retention behavior on the HLB columns, so that the fluctuation of recovery rate is large and the reproducibility is poor. In particular, the high salt and high pH characteristics of processed egg products (e.g., preserved eggs, salted eggs) further increase the difficulty of cleaning. Therefore, developing a specific purification method which can be applied to the chemical characteristics of the Hainanmycin of poultry meat, poultry eggs and complex egg product matrixes and is suitable for various complex matrixes has become a technical problem to be solved in the field. Disclosure of Invention In order to solve the technical problems, the invention provides a method for detecting the residue of Hainanmycin in poultry meat, eggs and egg products and a pretreatment method, and the method is suitable for detecting the Hainanmycin in poultry muscle, poultry liver, fresh eggs, liquid egg products, dried egg products, ice egg products and reproduced eggs, wherein the detection limit of the Hainanmycin is 0.2 mug/kg, and the quantitative limit of the Hainanmycin is 0.5 mug/kg. In order to achieve the above