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Method for determining amount of residual florfenicol in aquatic product by using high-efficiency liquid chromatogram fluorescence method

A technology of high performance liquid chromatography and florfenicol, which is applied in the field of detection of florfenicol residues in aquatic products

Inactive Publication Date: 2010-02-03
吴光红 +2
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0003] At present, in the domestic monitoring methods for detecting florfenicol residues, gas chromatography, high performance liquid chromatography-ultraviolet detector detection or chromatography-mass method are mostly used for detection, while high performance liquid chromatography-fluorescence detector is used to detect fluorobenzene The remnants of Nikau are still in a blank stage in China

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  • Method for determining amount of residual florfenicol in aquatic product by using high-efficiency liquid chromatogram fluorescence method
  • Method for determining amount of residual florfenicol in aquatic product by using high-efficiency liquid chromatogram fluorescence method
  • Method for determining amount of residual florfenicol in aquatic product by using high-efficiency liquid chromatogram fluorescence method

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Experimental program
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Effect test

Embodiment 1

[0047] Example 1: Extraction

[0048] Since the different ratios of acetonitrile and water will affect the retention time of the chromatographic peaks, in order to ensure that the peak time of Florfenicol is ideal and the peak shape is sharp, the ratio of acetonitrile and water in the mobile phase is finally determined as 25:75. The peak eluting time is ideal, and the target peak and impurity peak can be completely separated. As increasing the proportion of mobile phase acetonitrile, although the analysis time can be shortened, the peak time of florfenicol is too early, and it is difficult to separate completely from the impurity peak ( figure 2 ). As reducing the proportion of acetonitrile, the peak time of florfenicol is later, and the analysis time is longer ( image 3 ).

[0049] Therefore, in order to ensure that each target has a good separation effect and chromatographic peak shape, this method uses acetonitrile+water (25:75) solution as the mobile phase, and adopts...

Embodiment 2

[0050] Example 2: Purification

[0051] Many documents are processed to degreasing and purification, then concentrate, constant volume, and test on the machine. This study has also done a lot of experiments in this area, and found that the presence of some substrates interfered with the qualitative and quantitative peaks of the target drug and reduced the detection sensitivity when detecting low-concentration drugs ( Figure 4 ). Therefore, it is not enough to degrease and purify only by n-hexane, because n-hexane only removes most of the non-polar impurity components, and many polar substances, such as phospholipids, glycoproteins, etc., still exist in the extraction solvent, so further SPE purification is required . On the basis of comparing different types, solid-phase extraction columns of different types of fillers and different filler amounts on the purification effect of the target compound, it is concluded that the purification effect of the C18 solid-phase extractio...

Embodiment 3

[0052] Example 3: Method linear range and correlation

[0053] Take a certain amount of 1.0mg / L standard intermediate solution, dilute it with acetonitrile in sequence to 0.2, 0.5, 5, 10, 50, 100mg / L standard working solution, and detect according to the above chromatographic conditions. Carry out linear regression analysis to the mass concentration X (mg / L) of standard working solution with the peak area Y of each concentration, linear regression equation Y=225043X+82815, correlation coefficient r=0.9996 ( Figure 6 ). The results showed that: Florfenicol had a good linear relationship in the concentration range of 0.2μg / mL~100μg / mL.

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Abstract

The invention provides a method for determining the amount of residual florfenicol in an aquatic product by using a high-efficiency liquid chromatogram fluorescence method. In the method, florfenicolin an aquatic product sample is extracted by ethyl acetate, degreased by normal hexane, concentrated and purified by C18 solid extraction column, determined by a liquid chromatograph and a fluorescence detector and quantitated by an external standard method. The method can determine the residual florfenicol in the aquatic product quickly, sensitively and accurately.

Description

technical field [0001] The invention relates to a detection technology for detecting the residual amount of florfenicol in aquatic products. Background technique [0002] In the aquaculture industry, the diseases of aquatic animals often occur, especially the harm of bacterial diseases to aquatic animals is still very serious. The economic loss caused by the occurrence of diseases reaches tens of billions of dollars every year. Huge losses have been brought to the farming industry. In the past, chloramphenicol and other antibacterial drugs were often used to prevent and treat such diseases, but chloramphenicol is prone to drug resistance, and its disadvantage of causing aplastic anemia has caused my country's aquatic product exports to be repeatedly restricted and blocked. In 1996, the United States and the European Union, and in March 2002 in my country, banned the use of chloramphenicol in food animals. In view of the above situation, it is urgent to seek an alternative...

Claims

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Application Information

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IPC IPC(8): G01N30/06G01N21/64G01N30/74
Inventor 吴光红杨洪生陈玲珍孟勇张美琴
Owner 吴光红
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