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Streptomycin sulfate detection method

A technology of streptomycin sulfate and detection method, which is applied in the direction of chemical reaction of materials for analysis, chemiluminescence/bioluminescence, etc., can solve the problems of difficulty in obtaining, harsh environmental conditions, and long antibody preparation period, and achieves cost Low, highly tolerated effect

Inactive Publication Date: 2015-04-29
INST OF AGRO FOOD SCI & TECH CHINESE ACADEMY OF AGRI SCI
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  • Abstract
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  • Application Information

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Problems solved by technology

For example, the patent CN 103645322 A relates to a chemiluminescent enzyme-linked immunoassay kit for streptomycin sulfate, which includes an antigen, a streptomycin sulfate monoclonal antibody, and a chemiluminescent substance. detection, but the core material antibody of this detection method usually has a long preparation period, is difficult to obtain, and has strict requirements on environmental conditions such as temperature, humidity, and solvent properties.

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Experimental program
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Embodiment 1

[0045] Embodiment one: the pretreatment of sample

[0046] Taking honey as an example, the following sample pretreatments can be performed:

[0047] 1) Extraction: Homogenize 10g honey and mix with 20mL 0.1% phosphoric acid solution for 5 minutes;

[0048] 2) Purification: SCX solid phase extraction column (500mg / 3cc) and C18 solid phase extraction column (500mg / 6cc) dual column purification: a) Activation of SCX solid phase extraction column: add 5mL methanol and 5mL water to SCX solid phase extraction column, discard the effluent; b) SCX solid-phase extraction column loading: inject the extracted sample into the SCX solid-phase extraction column, and discard the effluent; c) SCX solid-phase extraction column rinsing: successively use 5mL 0.1 % phosphoric acid solution, 5mL water rinse, and the effluent was discarded; d) SCX solid-phase extraction column elution: 30mL 0.2mol / L dipotassium hydrogen phosphate solution (phosphoric acid to adjust pH 8.0), elute, collect the elua...

Embodiment 2

[0050] Embodiment 2: streptomycin sulfate detection method

[0051] Dosing:

[0052] Solution A: Weigh 12.5 mg of 9,10-phenanthrenequinone and dissolve it in a small amount of anhydrous ether, and dilute to volume in a 25 mL brown volumetric flask to prepare a 500 ppm solution A.

[0053] Solution B: Dissolve 400mg of sodium hydroxide in deionized water, make it volume in a 10mL volumetric flask, and prepare 1N sodium hydroxide aqueous solution B.

[0054] Standard solution: Prepare a series of streptomycin sulfate aqueous solutions such as 0ppm, 0.01ppm, 0.05ppm, 0.2ppm, 1ppm, 5ppm, 25ppm, and 100ppm with deionized water as standard solutions.

[0055] Sample solution to be tested: the sample solution to be tested obtained in Example 1 was used.

[0056] Add 0.5mL of solution A, 0.1mL of solution B and 0.4mL of streptomycin sulfate standard solution to a 1.5mL centrifuge tube in turn, shake fully for 30s, and let stand for 1min. In a new centrifuge tube, bright yellow-gree...

Embodiment 3

[0058] Embodiment 3: add recovery experiment

[0059] The method of the present invention is carried out sample addition recovery experiment, under the sample addition level of 100ppb, 500ppb, 1ppm, the recovery rate is respectively 82.3%, 88.5%, 85.3%, and the result shows that this method can meet the requirements applicable to streptomycin sulfate in food. Rapid screening for toxins.

[0060] The core material of the chemical fluorescent probe used in the present invention is chemical synthesis, with low cost and high tolerance to external environmental conditions. The streptomycin sulfate detection method is fast, convenient, sensitive and efficient, low in cost, does not need to perform complicated procedures of traditional methods, and is suitable for rapid screening of streptomycin sulfate in food.

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Abstract

The invention discloses a streptomycin sulfate detection method. The method is implemented through a specific reaction of a chemical fluorescent probe with streptomycin sulfate and comprises steps as follows: 1) dissolving the chemical fluorescent probe in an organic solvent to obtain a solution A, and dissolving alkali in water to obtain a solution B; 2) dissolving the streptomycin sulfate in water to prepare a series of streptomycin sulfate aqueous solutions with different concentrations, that is, standard solutions; 3) mixing and oscillating the solution A and the solution B with the standard solutions with the different concentrations respectively, leaving the mixtures to stand for layering, transferring water layers, measuring the fluorescence intensity, and establishing a correlation curve about the fluorescence intensity and the concentrations of the standard solutions; 4) mixing and oscillating the solution A and the solution B with a to-be-detected sample solution, leaving the mixture to stand for layering, transferring a water layer, measuring the fluorescence intensity, comparing the curve with the curve established in Step 3), and calculating the content of the streptomycin sulfate in a sample.

Description

technical field [0001] The invention belongs to the technical field of food safety analysis, relates to a detection method of streptomycin sulfate, in particular to a detection method of streptomycin sulfate based on an organic small molecule fluorescent probe. Background technique [0002] Streptomycin sulfate is a commonly used broad-spectrum aminoglycoside antibiotic, but it has certain toxic and side effects, mainly manifested as damage to the cranial nerves, hearing and kidneys. Therefore, many countries and institutions have clearly stipulated the maximum residue limits (MRLs) of such drugs in food. [0003] At present, the commonly used detection methods of streptomycin sulfate are mainly high-performance liquid chromatography (HPLC), chromatography-mass spectrometry (LC-MS) and enzyme-linked immunosorbent assay. The streptomycin sulfate instrument analysis method has high accuracy, but its operation is cumbersome, and derivatization is required, which is expensive, ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N21/76
Inventor 王淼陈卫华范蓓李敏敏黄亚涛孙玉凤卢嘉王凤忠
Owner INST OF AGRO FOOD SCI & TECH CHINESE ACADEMY OF AGRI SCI
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