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Method for detecting residual quantity of ivermectin in sheep muscle tissues by using liquid chromatograph/mass spectrometer with doramectin as internal standard substance

A muscle tissue, LC/MS technology, applied in measurement devices, instruments, scientific instruments, etc., can solve the problems of ivermectin loss, complicated and tedious operation procedures, etc., and achieves low detection limit, low coefficient of variation, and high sensitivity. Effect

Inactive Publication Date: 2015-12-23
LANZHOU INST OF ANIMAL SCI & VETERINARY PHARMA OF CAAS
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Acetonitrile-saturated n-hexane liquid-liquid extraction method, ivermectin is slightly soluble in n-hexane, and a certain amount of ivermectin will be lost while removing impurities
GB / T21320-2007 chooses C 18 and C 8 The solid phase extraction column is double purified, and the operation procedure is complicated and cumbersome

Method used

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  • Method for detecting residual quantity of ivermectin in sheep muscle tissues by using liquid chromatograph/mass spectrometer with doramectin as internal standard substance
  • Method for detecting residual quantity of ivermectin in sheep muscle tissues by using liquid chromatograph/mass spectrometer with doramectin as internal standard substance
  • Method for detecting residual quantity of ivermectin in sheep muscle tissues by using liquid chromatograph/mass spectrometer with doramectin as internal standard substance

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Experimental program
Comparison scheme
Effect test

Embodiment 1

[0045] Example 1 Effect of different ion source desolvation temperatures on the form of ion adducts

[0046] The ESI ion source is a soft ionization method, and ivermectin is usually combined with H in the ESI ion source + , Na + ,NH 4 + are combined to form quasi-molecular ion peaks, where [M+Na] + The ion structure is stable, if it is cracked by secondary mass spectrometry, it is not easy to obtain fragment ions, and the H + ,NH 4 + etc. can compete with the target molecule, thereby causing [M+Na] +The ion abundance is extremely unstable, so [M+NH 4 ] + as the parent ion.

[0047] 1. Conditions of liquid chromatography and mass spectrometry

[0048] 1. Liquid chromatography conditions

[0049] Analytical column: Agilent ZORBAX EclipsePlusC 8 Column; mobile phase A: acetonitrile, mobile phase B: 10 mmol / L ammonium acetate aqueous solution containing 0.1% formic acid (volume percentage); flow rate: 0.2 mL / min; column temperature: 25 °C; injection volume: 5 μL.

...

Embodiment 2

[0056] The standard curve assay experiment of embodiment 2 ivermectin

[0057] 1. Experimental steps

[0058] (1) Preparation of standard solution

[0059] 1. Preparation of doramectin internal standard solution

[0060] (1) Preparation of standard stock solution: Accurately weigh 10 mg of doramectin standard substance, dissolve and dilute it with acetonitrile, prepare a 100 μg / mL standard stock solution, and store it in a -20°C refrigerator in the dark.

[0061] (2) Preparation of standard solution: Accurately pipette 1 mL of doramectin standard stock solution into a 100 mL volumetric flask, dilute to the mark with acetonitrile and mix well to obtain a 1 μg / mL standard solution, store at 4°C for later use.

[0062] 2. Preparation of ivermectin standard solution

[0063] (1) Preparation of standard stock solution: Accurately weigh 10 mg of ivermectin standard substance, dissolve and dilute it with acetonitrile, prepare a 100 μg / mL standard stock solution, and store it in ...

Embodiment 3

[0083] The methodological verification of the residual determination of embodiment 3 ivermectin

[0084] By investigating method sensitivity, repeatability and recovery rate indicators, the method of the present invention is verified methodologically.

[0085] 1. Sensitivity experiment

[0086] According to embodiment 2 steps (two) to (six) process and analyze and measure the added sheep muscle sample, obtain the baseline noise of the added mark sheep muscle in the ivermectin retention time range, detect according to the calculation method of 3 times signal-to-noise ratio method Out of the limit, 10 times the signal-to-noise ratio method to calculate the quantitative limit. The test results show that the detection limit of the method of the present invention is 0.3 μg / kg, and the quantification limit is 0.5 μg / kg. The limits of ivermectin residues in sheep muscle in the United States, the European Union and my country are shown in Table 2. The detection limit and quantific...

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Abstract

The invention discloses a method for detecting the residual quantity of ivermectin in sheep muscle tissues by using a liquid chromatograph / mass spectrometer with doramectin as an internal standard substance. The method comprises the following steps: extracting ivermectin residual in the sheep muscle tissues with acetonitrile as an extraction solvent, adopting an impurity adsorption and solid phase extraction mode, adopting an alkaline alumina column as a solid phase extraction column, introducing acetonitrile, eluting, and collecting all obtained eluate. The method solves the problems of complex extraction and purification processes and high detection limit of traditional detection methods, realizes enrichment and purification of ivermectin residual in the sheep muscle tissues, improves the detection sensitivity, repeatability and accuracy, and is suitable for batch sample detection.

Description

technical field [0001] The invention belongs to the field of detection methods for veterinary drug residues, in particular to a method for detecting residual ivermectin in sheep muscle tissue. Background technique [0002] Ivermectin (IVM for short) is a new macrolide antibiotic, the second-generation derivative of Avermectins, which has the same deworming mechanism as Avermectin, both It is achieved by strengthening the effect of r-aminobutyric acid (GABA). GABA is an inhibitory neurotransmitter. In the brain, GABA mainly inhibits post-synaptic nerve conduction, and the release of GABA increases, which is the normal rest of post-synaptic cells. Potential energy is improved, it is difficult for nerves to transmit stimulation to muscles, muscles cannot contract, and parasites are paralyzed and expelled. It is one of the best anthelmintics in the abamectin family and is widely used in animal husbandry production. Ivermectin is widely used as an antiparasitic and acaricide in ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N30/02
Inventor 张继瑜周绪正李冰文豪魏小娟牛建荣程富胜王娟娟
Owner LANZHOU INST OF ANIMAL SCI & VETERINARY PHARMA OF CAAS
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