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Time-resolved fluorescent test strip for detecting profenofos and its application

A technology of time-resolved fluorescence and profenofos, applied in measuring devices, analytical materials, instruments, etc., can solve the problems of unsuitable basic laboratory batches, rapid detection of samples, difficult batch detection of samples, complex detection steps, etc., to achieve easy The effects of promotional use, high sensitivity, and short detection time

Pending Publication Date: 2017-11-24
联合益康(北京)生物科技有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The accuracy of the instrument detection method is high, but the instrument is expensive, requires professional technicians, the detection steps are complicated, and it is difficult to detect samples in batches. It is not suitable for batch and rapid detection of samples in grassroots laboratories.

Method used

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  • Time-resolved fluorescent test strip for detecting profenofos and its application
  • Time-resolved fluorescent test strip for detecting profenofos and its application
  • Time-resolved fluorescent test strip for detecting profenofos and its application

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0039] The preparation of embodiment 1 profenofos detection test strip

[0040] The preparation method of this test strip mainly comprises the following steps:

[0041] 1) Prepare a sample absorption pad sprayed with profenofos monoclonal antibody-time-resolved fluorescent marker;

[0042] 2) preparing a reaction membrane with a detection line coated with profenofos hapten-carrier protein conjugate and a quality control line coated with anti-rabbit antibody;

[0043] 3) Assemble the sample absorbent pad, reaction membrane and water absorbent pad prepared in 1) and 2) into a test strip.

[0044] The following step-by-step detailed description:

[0045] 1. Preparation of profenofos hapten

[0046] (1) Pour 20ml of absolute ethanol into a single-necked bottle, stir, and at ≤0°C, slowly add 5g of phosphorus trichloride dropwise, react for 30min, add 20ml of water, transfer to a separatory funnel, extract, and separate layers. The lower organic phase was removed, dried over anh...

Embodiment 2

[0070] The detection of profenofos residue in the sample of embodiment 2

[0071] 1. Sample processing

[0072] (1) Apple pretreatment method.

[0073] Homogenize the sample with a homogenizer; weigh (2.00±0.05)g homogenized apple sample into a 10mL polystyrene centrifuge tube, add 4mL 0.01M PBS, 2mL methanol, and vortex for 5min with a vortexer; room temperature Centrifuge at 4000r / min for 5min; take 200μL of the supernatant, add it to 800μL of 0.01M PBS, mix thoroughly, and take 50μL of it for analysis.

[0074] (2) Corn pretreatment method.

[0075] Homogenize the sample with a homogenizer; weigh (2.00±0.05)g homogenized corn sample into a 10mL polystyrene centrifuge tube, add 4mL 10% sodium chloride and 2mL methanol respectively, and vibrate with a vortexer for 5min; Centrifuge at 4000r / min for 5min at room temperature; take 100μL of the supernatant, add it to 900μL of 0.01M PBS, mix well, and take 50μL of it for analysis.

[0076] (3) Vegetable pretreatment method.

...

Embodiment 3

[0085] Example 3 sample detection example

[0086] 1. Detection limit test

[0087] Take blank samples of apple / corn / cucumber / tomato / radish leaf / cabbage, etc., add profenofos to the final concentration of 0.5ng / ml, 1ng / ml, 5ng / ml, 10ng / ml, 15ng / ml, 25ng / ml , 30ng / ml, 35ng / ml, take a test strip for detection, and repeat the measurement three times for each sample.

[0088] When testing samples such as apple / corn / cucumber / tomato / radish leaves / cabbage with test strips, the detection limit range is determined according to the test strip display results, indicating that the detection limit range of this test strip is 1ng / ml-30ng / ml.

[0089] 2. Sample precision and accuracy test

[0090] The recovery rate was used as the accuracy evaluation index, and the relative standard deviation (RSD%) of the test results of repeated determination of a certain concentration sample was used as the precision evaluation index. The calculation formula is: recovery rate (%)=actual measured value / ...

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Abstract

The invention discloses a time-resolved fluorescent test strip for detecting profenofos and its application. The test strip comprises a sample absorption pad, a reaction membrane and a water absorption pad, wherein the reaction membrane is located at the middle; the sample absorption pad and the water absorption pad respectively overlap the left and right ends of the reaction membrane; the sample absorption pad is provided with a sample loading zone and a microsphere zone; the microsphere zone are loaded bromofos time-resolved fluorescent microspheres; and the reaction membrane is provided with a test line ( a T line) and a control line (a C line), the T line is coated with a profenofos antigen (a coating antigen), and the C line coated with an anti-rabbit antibody. The invention also provides a method for detecting profenofos residues in samples of apples, corn, cucumbers, tomatoes, radish leaves, Chinese cabbages and the like by using the profenofos test strip. The test strip provided by the invention has the advantages of simple operation, high sensitivity, quantitative detection, rapid detection, low cost and the like, and is suitable for screening and on-site monitoring of a large number of samples.

Description

technical field [0001] The invention relates to a time-resolved fluorescent test strip for detecting profenofos and its application, which belongs to the technical field of time-resolved fluorescent immunoassay (TRFIA), and is used for profenofos in samples such as apples, corns, cucumbers, tomatoes, radish leaves, and cabbage. Content detection of bromphos. Background technique [0002] Profenofos is an organophosphorus insecticide, also known as bromochlorphos and polyfenphos, which can inhibit pests on vegetables, cotton and other crops, and is mainly suitable for the control of cotton bollworm, cotton aphid, pink bollworm, Rice leaf roller, etc. Profenofos is a light yellow liquid, miscible with most organic solvents, relatively stable in neutral and slightly acidic conditions, unstable in alkaline environments. Profenofos has contact and stomach toxicity, and the action is rapid. As the pesticide is widely used, it will inevitably cause pollution to vegetables and ot...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/558G01N33/533
CPCG01N33/533G01N33/558
Inventor 吴鹏姚承志王新华
Owner 联合益康(北京)生物科技有限公司
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