Method for detecting veterinary drug residues in meat product

Abstract

The invention provides a method for detecting veterinary drug residues in meat products, which comprises the following steps: S1, quantitatively taking 5g of sample, cutting, grinding, transferring into a 50ml centrifugal tube, adding 20ml of EDTA-Na solution, carrying out ultrasonic treatment for 15-20 minutes, then carrying out low-temperature high-speed centrifugal treatment, collecting supernatant liquid, adding 20ml of EDTA-Na solution into lower-layer precipitate again, and carrying out ultrasonic treatment for 15-20 minutes; performing ultrasonic treatment and low-temperature high-speed centrifugal treatment, repeating for 2-5 times, and combining all supernatant liquid; s2, all the supernate in the S1 passes through an SPE small column filled with macroporous resin, then pure water is used for leaching the SPE small column, then a methanol solution with the volume ratio being 85% is used for leaching the SPE small column, and purified liquid is obtained; and S3, concentrating the purified liquid obtained in S2 to a constant volume of 1 ml, and then carrying out liquid chromatography-gas chromatography-mass spectrometry detection. The method solves the problem that the efficiency is low when the veterinary drug residues in the meat product are detected by liquid chromatography-gas chromatography-mass spectrometry in the prior art.

Description

Technical Field

[0001] The invention relates to the technical field of veterinary drug residue detection, and in particular to a method for detecting veterinary drug residues in meat products. Background Art

[0002] In the process of livestock breeding, in order to prevent livestock from getting sick, veterinary drugs are inevitably used, such as the use of tetracycline antibiotics in pig feed. Using less or disabling veterinary drugs before slaughtering livestock can effectively prevent excessive veterinary drugs in livestock meat. Livestock meat (such as pork) is also processed into various side dishes, such as ham, steamed buns, meat floss cakes, etc. During the processing, the use of various additives is also involved. Therefore, it is more difficult to detect veterinary drugs in side dishes and is affected by more impurity factors. In the prior art, more advanced detection technologies can be used for detection, such as liquid-gas mass spectrometry detection, which can achieve better results. However, complex pre-treatment is required, which makes the overall detection process complicated. Summary of the invention

[0003] In view of the deficiencies in the prior art, the present invention provides a method for detecting veterinary drug residues in meat products, which solves the problem of low efficiency in the prior art when using liquid-gas mass spectrometry to detect veterinary drug residues in meat products.

[0004] According to an embodiment of the present invention, a method for detecting veterinary drug residues in meat products comprises the following steps:

[0005] S1, quantitatively take 5g of sample, cut and grind it, transfer it to a 50ml centrifuge tube, add 20ml EDTA-Na solution, ultrasonicate it for 15-20min, and then perform low-temperature high-speed centrifugation, collect the upper clear liquid, add 20ml EDTA-Na solution to the lower precipitate again, perform ultrasonicate, low-temperature high-speed centrifugation, repeat 2-5 times, and combine all the upper clear liquid;

[0006] S2, all the supernatants in S1 are passed through a SPE column filled with a macroporous resin, and then the SPE column is eluted with pure water, and then the SPE column is eluted with a methanol solution with a volume ratio of 85% to obtain a purified solution;

[0007] S3: The purified liquid in S2 is concentrated to 1 ml and then detected by liquid chromatography-mass spectrometry.

[0008] Furthermore, the concentration of the EDTA-Na solution is 0.1 mol / L.

[0009] Furthermore, the low-temperature high-speed centrifugation treatment conditions are: 4°C, a rotation speed of 13000r / min, and centrifugation for 8-10min.

[0010] Furthermore, the pH value of the supernatant in S1 is adjusted to weak acidity before performing step S2.

[0011] Furthermore, the pH of the methanol solution is 2.

[0012] Furthermore, the purified liquid in S2 is also filtered.

[0013] Furthermore, the ultrasonic treatment power is 400-450w.

[0014] Furthermore, the liquid chromatography conditions were as follows: BEH-C18 column, column temperature 35°C, mobile phase A was 0.1% formic acid solution, phase B was acetonitrile, detection was performed by gradient elution, flow rate 0.3 mL / min, injection volume 10 uL.

[0015] Furthermore, the mass spectrometry conditions were: electrospray ion source, positive ion scanning, multiple reaction detection, ionization voltage 1.0 kV, ion source temperature 150°C, nebulization temperature 400°C, cone gas flow rate and nebulizer flow rate 50 L / h and 800 L / h, respectively.

[0016] Furthermore, the SPE cartridge is subjected to activation treatment.

[0017] Compared with the prior art, the present invention has the following beneficial effects:

[0018] Through ultrasonic treatment, the extraction efficiency of veterinary drug residues in the sample is improved, and then the target object is extracted more efficiently in the subsequent solid phase extraction process. The overall pretreatment time is short, which solves the problem of low efficiency in the existing technology of using liquid-gas mass spectrometry to detect veterinary drug residues in meat products. DETAILED DESCRIPTION

[0019] The technical solution of the present invention is further described below in conjunction with embodiments.

[0020] This embodiment provides a method for detecting veterinary drug residues in meat products, which improves the extraction efficiency of veterinary drug residues by ultrasonic treatment, and specifically includes the following steps:

[0021] Quantitatively sample 5g, cut and grind, transfer to a 50ml centrifuge tube, add 20ml EDTA-Na (pH 5) solution of 0.1mol / L, ultrasonically treat for 15-20min at 400-450w, then perform low-temperature high-speed centrifugation at 4℃ and 13000r / min for 8-10min, collect the upper clear liquid, add 20ml of the above EDTA-Na solution to the lower precipitate again, perform ultrasonic treatment and low-temperature high-speed centrifugation, repeat 2-5 times, combine all the upper clear liquids, adjust the pH of all the supernatants to 4, then pass through the SPE column filled with macroporous resin (after activation treatment), and then rinse the SPE with pure water. The SPE column was then eluted with a methanol solution with a volume ratio of 85% and a pH of 2 to obtain a purified liquid, which was filtered through a 0.22 μm membrane and then concentrated to 1 ml. The liquid chromatography conditions were as follows: BEH-C18 column, column temperature of 35°C, mobile phase A of 0.1% formic acid solution, phase B of acetonitrile, detection by gradient elution, flow rate of 0.3 mL / min, injection volume of 10 uL; mass spectrometry conditions were as follows: electrospray ion source, positive ion scanning, multiple reaction detection, ionization voltage of 1.0 kV, ion source temperature of 150°C, atomization temperature of 400°C, cone gas flow rate and nebulizer flow rate of 50 L / h and 800 L / h, respectively.

[0022] The following table lists the comparison results of specific embodiments:

[0023]

[0024]

[0025] Finally, it should be noted that the above embodiments are only used to illustrate the technical solution of the present invention rather than to limit it. Although the present invention has been described in detail with reference to the preferred embodiments, those skilled in the art should understand that the technical solution of the present invention can be modified or replaced by equivalents without departing from the purpose and scope of the technical solution of the present invention, which should be included in the scope of the claims of the present invention.

Claims

1. A method for detecting veterinary drug residues in meat products, characterized in that: The steps include: S1: quantitatively take 5g of sample, chop and grind it, transfer it to a 50ml centrifuge tube, add 20ml of EDTA-Na solution, ultrasonicate it for 15-20min, and then centrifuge it at low temperature and high speed. Collect the supernatant, add 20ml of EDTA-Na solution to the lower precipitate, ultrasonicate it again, and centrifuge it at low temperature and high speed. Repeat 2-5 times, and combine all the supernatants. All supernatants in S2 and S1 are passed through an SPE column filled with macroporous resin, and then the SPE column is eluted with pure water, and then the SPE column is eluted with a methanol solution with a volume ratio of 85% to obtain the purified solution; S3. The purified solution in S2 was concentrated to 1 ml and then detected by liquid chromatography-gas chromatography-mass spectrometry.

2. The method for detecting veterinary drug residues in meat products according to claim 1, wherein: The concentration of EDTA-Na solution is 0.1 mol / L.

3. The method for detecting veterinary drug residues in meat products according to claim 1, wherein: The low-temperature high-speed centrifugation treatment conditions are: 4°C, a rotation speed of 13000 r / min, and centrifugation for 8-10 minutes.

4. The method for detecting veterinary drug residues in meat products according to claim 1, wherein: The pH value of the supernatant in S1 is adjusted to weak acidity before proceeding to step S2.

5. The method for detecting veterinary drug residues in meat products according to claim 1, wherein: The pH of the methanol solution is 2.

6. The method for detecting veterinary drug residues in meat products according to claim 1, wherein: The purified liquid in S2 is further filtered.

7. The method for detecting veterinary drug residues in meat products according to claim 1, wherein: The ultrasonic treatment power is 400-450w.

8. The method for detecting veterinary drug residues in meat products according to claim 1, wherein: Liquid chromatography conditions were as follows: BEH-C18 column, column temperature 35°C, mobile phase A consisting of 0.1% formic acid solution, mobile phase B consisting of acetonitrile, detection by gradient elution, flow rate 0.3 mL / min, injection volume 10 μL.

9. The method for detecting veterinary drug residues in meat products according to claim 1, wherein: The mass spectrometry conditions were as follows: electrospray ion source, positive ion scanning, multiple reaction detection, ionization voltage 1.0 kV, ion source temperature 150 °C, nebulization temperature 400 °C, cone gas flow rate and nebulizer flow rate 50 L / h and 800 L / h, respectively.

10. The method for detecting veterinary drug residues in meat products according to any one of claims 1 to 9, characterized in that: The SPE cartridge was activated.

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