阅读说明：本技术 一种兽药喹烯酮中乙酰甲喹的检测方法 (Method for detecting mequindox in veterinary drug quinocetone ) 是由 郝燕娟 王智民 黄妍 洪思悦 曾敏 刘丽英 于 2020-04-30 设计创作，主要内容包括：本发明公开了一种兽药喹烯酮中乙酰甲喹的检测方法。乙酰甲喹样品经二甲亚砜溶解,甲醇超声提取后,再经C18色谱柱分离,利用高效液相色谱仪进行定性分析,采用外标法定量。本发明的检测方法步骤简单,易于操作,准确度、精密度和灵敏性高,可作为制定兽药喹烯酮中乙酰甲喹检测标准的参考依据。(The invention discloses a method for detecting mequindox in veterinary drug quinocetone. Dissolving mequindox sample in dimethyl sulfoxide, ultrasonically extracting with methanol, separating with C18 chromatographic column, performing qualitative analysis with high performance liquid chromatograph, and quantifying with external standard method. The detection method disclosed by the invention is simple in steps, easy to operate and high in accuracy, precision and sensitivity, and can be used as a reference basis for formulating the detection standard of mequindox in veterinary drug quinocetone.)

1. A detection method of mequindox in veterinary drug quinocetone is characterized by comprising the following steps:

(1) adding dimethyl sulfoxide into a quinocetone sample for ultrasonic dissolution, adding methanol for ultrasonic extraction, cooling, performing constant volume with methanol, shaking up, and filtering to obtain a test solution;

(2) and (3) carrying out high performance liquid chromatography detection on the test solution, carrying out qualitative detection according to the retention time of a chromatographic peak, and carrying out quantitative detection by using a peak area external standard method.

2. The method for detecting mequindox in veterinary quinocetone according to claim 1, wherein in the step (2), the chromatographic conditions are specifically as follows:

a chromatographic column: a C18 chromatography column;

the mobile phase comprises the following components in percentage by volume: acetonitrile: 20 parts of water: 80;

flow rate: 1.0 mL/min;

column temperature: 30 ℃;

detection wavelength: 257 nm;

sample introduction amount: 10 μ L.

3. The method for detecting mequindox in veterinary quinocetone according to claim 2, wherein the chromatographic column is Hypersil GOLD aQ, and the specification is as follows: 4.6X 250mm, 5 μm.

4. The method for detecting mequindox in veterinary drug quinocetone according to claim 1, wherein in the step (1), the ratio of the quinocetone sample to the dimethyl sulfoxide is 1g:20 mL.

5. The method for detecting mequindox in veterinary quinocetone according to claim 1, wherein in the step (1), the time for ultrasonic extraction is not less than 10 min.

6. The method for detecting mequindox in veterinary quinocetone according to any one of claims 1-5, wherein in step (1), the test solution is filtered through a 0.45 μm filter.

Technical Field

The invention relates to the technical field of mequindox detection and analysis, and particularly relates to a method for detecting mequindox in veterinary drug quinocetone.

Background

Quinocetone (Quinocetone) is a national-grade new drug officially approved in 8 months in 2003 by Ministry of agriculture and is a new veterinary drug initiated internationally in our country. The composition has antibacterial, safe use, rapid metabolism, and obvious effect, and is suitable for preventing diseases and promoting growth of pig, fowl, aquatic product, young livestock, and young fowl. Zhang et al, in the research of the relationship between water solubility and in vitro antibacterial activity of different quinenone hydroxy derivatives, propose the synthetic method of the quinenone compound, namely take mequindox as raw materials, through condensation reaction with benzaldehyde substituted by hydroxy at different positions, get quinocetone or quinenone derivatives. Due to the problems of the process, the problem of residual quantity of mequindox serving as an intermediate of quinocetone is often ignored, and no means for detecting and limiting mequindox in veterinary quinocetone exists at present.

Disclosure of Invention

The invention aims to overcome the defects of the prior art and provides a method for detecting mequindox in veterinary drug quinocetone, which has simple steps and easy operation and can effectively determine the residual quantity of mequindox as an intermediate of quinocetone.

The chemical structural formula of mequindox is as follows:

in order to achieve the purpose, the technical scheme adopted by the invention is as follows:

a detection method of mequindox in veterinary drug quinocetone comprises the following steps:

(1) adding dimethyl sulfoxide into a quinocetone sample for ultrasonic dissolution, adding methanol for ultrasonic extraction, cooling, diluting to a constant volume with methanol, shaking up, and filtering;

(2) and (3) carrying out high performance liquid chromatography detection on the test solution, carrying out qualitative detection according to the retention time of a chromatographic peak, and carrying out quantitative detection by using a peak area external standard method.

The inventor selects different extraction modes to carry out pretreatment on the sample, and the result shows that the best effect is achieved by ultrasonic extraction with methanol after the dimethyl sulfoxide is added for ultrasonic dissolution. The detection method is simple and convenient to operate and rapid, and can effectively determine the intermediate mequindox remained in the veterinary drug quinocetone.

Further, in the step (2), the chromatographic conditions are specifically as follows:

a chromatographic column: a C18 chromatography column;

the mobile phase is acetonitrile according to the volume ratio: 20 parts of water: 80, selecting the acetonitrile water as a mobile phase, and separating mequindox from other component chromatographic peaks better;

flow rate: 1.0 mL/min;

column temperature: 30 ℃;

detection wavelength: 257 nm;

sample introduction amount: 10 μ L.

Under the condition of the liquid chromatography, the mequindox peak can be well separated and detected, and the accuracy, the precision density and the sensitivity are high.

Further, the chromatographic column is Hypersil GOLD aQ, and the specification is as follows: 4.6X 250mm, 5 μm, good separation effect was obtained.

Further, in the step (1), the ratio of the quinocetone sample to the dimethyl sulfoxide is 1g:20mL, so that excessive dimethyl sulfoxide is prevented from influencing the peak shape of the map.

Further, in order to ensure complete ultrasonic extraction, in the step (1), the time of ultrasonic extraction is more than or equal to 10 min.

Further, in the step (1), the sample solution is filtered through a 0.45 μm filter.

Compared with the prior art, the invention has the beneficial effects that:

the invention establishes a method for detecting mequindox in veterinary drug quinocetone, wherein a sample is subjected to ultrasonic dissolution by dimethyl sulfoxide, ultrasonic extraction by methanol, separation by a C18 chromatographic column, qualitative analysis by a high performance liquid chromatograph, and quantification by an external standard method. The method has simple steps, easy operation and high accuracy, precision and sensitivity, and can be used as a reference basis for establishing the detection standard of mequindox in quinocetone.

Drawings

FIG. 1 is a chromatogram of a control solution at 257nm at a mequindox concentration of 10. mu.g/mL.

FIG. 2 is a chromatogram of a test solution at 257 nm.

FIG. 3 is a spectrum of mequindox standard.

FIG. 4 is a spectrum of a sample.

FIG. 5 is a superimposed spectrum of mequindox standard and test sample.

Detailed Description

To better illustrate the objects, aspects and advantages of the present invention, the present invention will be further described with reference to specific examples. It will be understood by those skilled in the art that the specific embodiments described herein are merely illustrative of the invention and are not intended to limit the invention.