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Visualized detection method aiming at aquatic food fluoroquinolone analog drug residue

A fluoroquinolone and detection method technology, applied in the field of detection of drug residues in aquatic food, can solve the problems such as cumbersome, it takes several hours at the fastest, poor stability of detection results, difficult on-site detection, etc. Easy to carry and assemble and shorten the analysis time

Inactive Publication Date: 2008-07-02
OCEAN UNIV OF CHINA
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

It has a high degree of specificity and sensitivity, but the analysis procedure is relatively cumbersome, and it takes several hours at the fastest to report the test results. Operators need to have high professional skills and operating experience; enzymes are more sensitive to interference from external factors. The stability of the test results is poor; in addition, equipment such as a microplate reader is required, which generally needs to be carried out in a laboratory, and it is difficult to carry out on-site detection

Method used

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  • Visualized detection method aiming at aquatic food fluoroquinolone analog drug residue

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0028] Example 1: Rapid detection of enrofloxacin, ciprofloxacin and norfloxacin residues in eels

[0029] 1. Preparation of antibodies

[0030] First prepare phosphate buffer (0.01M, pH7.4, referred to as PBS): take NaCl 8g, KCl 0.2g, Na 2 HPO 4 ·12H 2 O 2.9g, KH 2 PO 4 2.0g, add double distilled water to 1000mL, pass through 0.22μm membrane, and store at room temperature.

[0031] The carbodiimide synthesis procedure is as follows: dissolve 15mg EF, 5.2mg NHS, 9.3mg EDC in 0.5mL dimethylformamide (DMF for short), shake at room temperature for 1.5h; centrifuge at 3000r / min for 30s, add 2mL supernatant The PBS solution containing 25 mg of carrier protein (bovine serum albumin) (pre-dissolved with 33% DMF) was shaken overnight at constant temperature (150 r / min, 4 °C); DMF was dialyzed against PBS solution, and the amount of DMF was gradually reduced until PBS was completely used.

[0032] Healthy BALB / C mice aged 6-8 weeks were selected for immune test. Blood was coll...

Embodiment 2

[0055] Embodiment 2: the detection effect of the present invention and its comparison with HPLC method

[0056] Table 1 Detection results of spiked eel meat by HPLC (n=3)

[0057] Add Concentration

[0058] At present, the requirements for fluoroquinolone drug residues in food at home and abroad vary according to different drug varieties and different food types, but the minimum detection limit is generally required to be above 50 μg / kg. Using the detection method of the present invention, when the added concentration of enrofloxacin and ciprofloxacin is 20ng / mL, it can be judged as positive by naked eyes, while the detection limit of norfloxacin is at 50ng / mL; The results are basically the same; the recovery rate of the pretreatment method can basically be guaranteed to be above 70%. This shows that the main performance of this method, such as sensitivity and accuracy, can fully meet the requirements of actual detection; but the detection time only takes 25-30 mi...

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Abstract

The invention discloses a visualized detection method of residual fluoroquinolones in sea foods. The method comprises the following steps of: preparing a specific antibody by coupling bovine serum albumin (used as a carrier protein) and fluoroquinolones with immunological technique; conjugating the specific antibody and red colloidal gold particles to obtain a colloidal gold-labeled antibody; coating and blocking a nitrocellulose membrane (used as a solid carrier) with another antibody; sequentially adding a sample solution, the colloidal gold-labeled antibody and a washing solution, and preparing a negative control from an extractive solution containing no fluoroquinolones; and compared with the negative control, and determining whether the sample is positive (containing fluoroquinolones) based on the existence of macroscopic red spots. The invention can be used for rapid field screening and detection of multiple residual fluoroquinolones; and has the advtanges of high sensitivity and accuracy, simple and rapid operation, no need of extra analytical apparatus, and macroscopic result.

Description

technical field [0001] The invention relates to a method for detecting drug residues in aquatic food, in particular to a method for detecting fluoroquinolone drug residues. The invention is funded by the National Natural Science Foundation of China (No. 30400336) and the Qingdao Science and Technology Project (03-1-NSH-2-2). Background technique [0002] Fluoroquinolones (FQs) are new synthetic broad-spectrum bactericidal antibacterial drugs, which have the advantages of broad antibacterial spectrum, strong antibacterial activity, less toxic and side effects, and less cross-resistance with other antibiotics and antibacterial drugs. Therefore, it is widely used in aquatic products and livestock and poultry breeding, and has become one of the commonly used fish and veterinary drugs in my country. Due to the large and long-term use of fluoroquinolones, fluoroquinolones accumulate in the body and cause drug residues in animal tissues, which is potentially harmful to food hygiene...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/543
Inventor 曹立民林洪隋建新李振兴江洁
Owner OCEAN UNIV OF CHINA
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