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Colloidal gold test paper card for detecting enrofloxacin medicament residue

The technology of colloidal gold test paper and enrofloxacin is applied in the field of immunochemical rapid detection of veterinary drug residues, which can solve the problems of being unsuitable for on-site monitoring and screening of a large number of samples, unfavorable for the promotion and use of grassroots units, and complex instruments and equipment, and achieves detection of The effect of short time, low cost and high sensitivity

Inactive Publication Date: 2009-08-05
贵州勤邦食品安全科学技术有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

One is chromatographic analysis, such as high performance liquid chromatography (HPLC), which is not suitable for on-site monitoring and screening of a large number of samples due to complex instruments and tedious processes
The other is an immunological method, such as enzyme-linked immunosorbent assay (ELISA), which has the disadvantages of long detection time and high cost, which is not conducive to popularization and use in grassroots units.
Moreover, both of these methods require professional and technical personnel to operate

Method used

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  • Colloidal gold test paper card for detecting enrofloxacin medicament residue
  • Colloidal gold test paper card for detecting enrofloxacin medicament residue
  • Colloidal gold test paper card for detecting enrofloxacin medicament residue

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0031] Example 1 Preparation of Enrofloxacin Detection Test Paper Card

[0032] 1. Synthesis and identification of enrofloxacin-carrier protein conjugates

[0033] (1) Synthesis of Enrofloxacin-Human Serum Protein Conjugate

[0034] 30mg of enrofloxacin was dissolved in 1ml of dimethylformyl. Cool to 10°C, add 40ul of isobutyl chloroformate, stir and react at 10°C for 30 minutes. 120mgHSA was dissolved with 2ml 50mmol / LNa2CO2. React at 10°C for 4 hours (add NaOH if necessary to maintain the pH of the solution at 9.0), then overnight at 4°C. Freeze-dry and store at -20°C.

[0035] (2) Synthesis of enrofloxacin-bovine serum albumin:

[0036] Take 100 mg of EDC and dissolve it fully with 1.5 mL of 10 mmol / L PBS solution with pH 8.0 (I solution). Take 40mg of enrofloxacin and dissolve it in 2ml of 0.2mol / L NaOH solution (II liquid). Take 100mg of BSA and dissolve it in 3ml of 10mmol / L PBS (pH8.0) solution (III solution).

[0037] Mix liquid II and liquid III, and add liqui...

Embodiment 2

[0063] Example 2 Detection of Enrofloxacin Residues in Samples

[0064] 1. Sample pretreatment

[0065] (1) Animal tissue pretreatment (chicken, chicken liver, pork, pork liver, fish, shrimp)

[0066] Weigh 1.0±0.05g of the homogenized tissue sample into a centrifuge tube, add 4ml of deionized water, and tightly cap the bottle. Place the centrifuge tube containing the sample in a slightly boiling water bath for 10 minutes, draw more than three drops of the solution and pour it into a 1.5ml centrifuge tube. If there is obvious yellow turbidity, please centrifuge, and then use the supernatant as the sample solution to be tested.

[0067] (2) Pretreatment of serum samples

[0068] Put a small amount of serum sample in a centrifuge tube, add 4ml of deionized water, and mix it as the sample solution to be tested.

[0069] 2, detect with the test paper card of the present invention

[0070] Use a dropper to drop 3 drops of sample into the hole of the reagent card, and observe t...

Embodiment 3

[0073] Example 3 Example of sample detection

[0074] Take 20 samples each of pork, chicken, pork liver, shrimp, and serum and 20 samples of various negative samples with known enrofloxacin drug residue concentration greater than 40ng / g, repeat the test twice for each sample, and calculate its negative and positive results Rate. The results are shown in Table 1 to Table 5.

[0075] Table 1 Negative and positive rate detection of pork samples

[0076]

[0077]

[0078] Table 2 Detection of negative and positive rate of chicken samples

[0079]

[0080]

[0081] Table 3 Negative and positive rate detection of pig liver samples

[0082]

[0083]

[0084] Table 4 Detection of negative and positive rate of shrimp samples

[0085]

[0086] Table 5 Negative and positive rate detection of serum samples

[0087]

[0088]

[0089] The results showed that pork, chicken, pork liver, shrimp, serum 20 positive samples and 20 respective negative samples, each ...

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Abstract

The invention provides a colloidal gold immunologic test paper card for detecting enrofloxacin drugs, comprising a reaction film, a sample pad, a conjugate releasing pad, a water absorbing pad and a back lining, wherein the reaction film is provided with a test zone coated with the enrofloxacin drugs, namely carrier protein conjugates and a quality control zone coated with sheep-anti-mouse IgG; and the conjugate releasing pad is coated with a monoclonal antibody of the enrofloxacin drugs, namely a colloidal gold label. The invention also provides a method for applying the test paper card to detect the enrofloxacin drugs, including the following steps of sample preliminary treatment, detection by using the test paper card and detected result analysis. The test paper card can be used for detecting the residual quantity of the enrofloxacin drugs in animal foods, such as pork, pork liver, chicken, chicken liver, serum, fishes and shrimps, has the advantages of simple operation, high sensitivity, rapid detection speed and low cost, and can monitor on site and satisfy the requirement of screening a plurality of samples.

Description

technical field [0001] The present invention relates to the technical field of immunochemical rapid detection of veterinary drug residues, and more specifically relates to a colloidal gold test paper card for detecting enrofloxacin drug residues, and rapid detection of enrofloxacin by means of immunochromatographic reaction of colloidal gold labeling and color development drug residue. Background technique [0002] Enrofloxacin (enrofloxaicn, ENR) drug is a class of very important broad-spectrum antibiotics developed rapidly in the past 20 years. In terms of chemical structure, this class of drugs belongs to pyridonecarboxylic acids (PCAs). Inhibit bacterial DNA helicase, broad antibacterial spectrum, high efficiency, low toxicity, strong tissue penetration ability, antibacterial effect is nearly a thousand times that of sulfonamides, comparable to third-generation cephalosporin antibiotics. Due to its chemical structure and low price, it will soon be widely used in medici...

Claims

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Application Information

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IPC IPC(8): G01N33/577G01N33/558
Inventor 何方洋沈建忠万宇平冯才伟冯才茂赵正苗吴小平朱亮汪善良
Owner 贵州勤邦食品安全科学技术有限公司
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