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Atrazine detection kit based on upconversion fluorescence immunoassay sensor, application and atrazine detection method

A detection kit and atrazine technology, applied in the field of food safety detection immunoassay, can solve problems such as the influence of complex components, complex detection procedures, sensitivity and accuracy defects, and achieve high specificity

Active Publication Date: 2018-09-14
INST OF ENVIRONMENTAL MEDICINE & OCCUPATIONAL MEDICINE ACAD OF MILITARY MEDICINE ACAD OF MILITARY SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Large instrument methods require expensive instrumentation, time-consuming sample pretreatment, and complex detection procedures
As a traditional immunoassay detection method, ELISA has been widely used in some areas, but in the detection process of actual samples (such as corn, sugarcane, etc.), it is easily affected by the complex components in food, which makes the method more sensitive and accurate in detection. Sexual defects still exist

Method used

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  • Atrazine detection kit based on upconversion fluorescence immunoassay sensor, application and atrazine detection method
  • Atrazine detection kit based on upconversion fluorescence immunoassay sensor, application and atrazine detection method
  • Atrazine detection kit based on upconversion fluorescence immunoassay sensor, application and atrazine detection method

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Experimental program
Comparison scheme
Effect test

Embodiment 1

[0046] 1. Preparation of anti-atrazine antibody labeled with core / shell upconversion particles

[0047] 1) Core / shell upconversion particles (NaYF 4 :Yb,Er@NaYF 4 :Nd) synthetic method

[0048] ①Wash all glassware with aqua regia, rinse thoroughly with deionized water, and then dry in a 100°C drying oven.

[0049] ②Preparation of rare earth element trifluoroacetic acid precursor: Accurately weigh 10mmol each of rare earth element oxides (yttrium oxide, ytterbium oxide, and erbium oxide) into three 20mL two-necked bottles, install a condensation reflux device, and fix them in an oil bath , and the temperature was raised to 80°C. Add 60mmol trifluoroacetic acid under slow magnetic stirring, keep the temperature for 24h. After the reaction, dry in a vacuum oven at 60° C. for 12 hours. The obtained rare earth trifluoroacetate is collected and stored in a constant temperature and humidity drying oven.

[0050] ③ Synthesis of upconversion particles (NaYF 4 : Yb, Er) inner core:...

Embodiment 2

[0070] Applied method of an upconverting fluorescent immunosensor for the detection of atrazine.

[0071] 1) Block 500 μL of antibody-modified upconversion particles (0.5 mg / mL) with 1% BSA blocking solution for 1 h at 37° C. Centrifuge at 11000 rpm for 5 min, discard the supernatant, wash repeatedly with 0.01 M PBS for 5 min each time, and redissolve in 500 μL of PBS.

[0072] 2) Add a certain concentration of diluted (0.01ng mL -1 , 0.1ng mL -1 , 1ng mL -1 , 10ng mL -1 , 100ng mL -1 , 1000ng mL -1 ) atrazine standard, add 500 μL, and incubate at 37° C. for 1 h with slow shaking.

[0073] 3) After the reaction, centrifuge at 11,000 rpm for 5 minutes to remove unconnected small molecules, wash repeatedly with PBS solution, and redissolve in 500 μL PBS.

[0074] 4) Add 500 μL of black phosphorous nano-gold (0.5 mg / mL) and shake vigorously for 10 min to form a structural complex (UCP-mAb@BP-Au).

[0075] 5) After the reaction, centrifuge at 11,000 rpm for 10 min, and was...

Embodiment 3

[0079] Specificity experiments of an upconverting fluorescent immunosensor for the detection of atrazine.

[0080] 1) Block 500 μL of antibody-modified upconversion particles (0.5 mg / mL) with 1% BSA blocking solution for 1 h at 37° C. Centrifuge at 11000 rpm for 5 min, discard the supernatant, wash repeatedly with 0.01 M PBS for 5 min each time, and redissolve in 500 μL of PBS.

[0081] 2) Add 500 μL (500 ng / mL) standard products of chlorpyrifos, diethylstilbestrol, simazine, melamine, and atrazine to different tubes respectively. Slowly shake the reaction at 37°C for 1 h.

[0082] 3) After the reaction, centrifuge at 11,000 rpm for 5 min to remove unconnected small molecules, wash repeatedly with PBS solution, and redissolve in 500 μL PBS.

[0083] 4) Add 500 μL of black phosphorous nano-gold (0.5 mg / mL) and shake vigorously for 10 min to form a structural complex (UCP-mAb@BP-Au).

[0084] 5) After the reaction, centrifuge at 11,000 rpm for 10 min, and wash with 500 μL of PB...

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Abstract

The invention belongs to the technical field of food safety detection immunoassay and relates to an atrazine detection kit based on an upconversion fluorescence immunoassay sensor, application and anatrazine detection method. The kit comprises carboxylation core / shell type upconversion nanoparticles NaYF4:Yb,Er@NaYF4:Nd, anti-atrazine monoclonal antibody and black phosphorus nanogold composite material dispersion liquid. By utilizing the kit or the detection method, high-sensitivity detection of atrazine can be realized.

Description

technical field [0001] The invention belongs to the technical field of food safety detection immunoanalysis, and more specifically relates to an atrazine detection kit based on an up-conversion fluorescent immunosensor and its application and an atrazine detection method based on an up-conversion fluorescent immunosensor . Background technique [0002] Atrazine, also known as atrazine, is a broad-spectrum herbicide that can control a variety of annual grasses and broad-leaved weeds. It is suitable for upland crops such as corn, sorghum, sugar cane, fruit trees, nurseries, and woodlands. . However, large-scale use of atrazine can contaminate food and drinking water through various channels, leading to serious health risks. The US Environmental Protection Agency reports that overdose of atrazine can trigger vital medical conditions such as low blood pressure, muscle cramps and adrenal damage. Currently, atrazine residues can be detected in both groundwater and surface water...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/577G01N33/543G01N33/533G01N21/64
CPCG01N21/6428G01N33/533G01N33/54346G01N33/577G01N2021/6432G01N2021/6439
Inventor 王江高志贤白家磊彭媛任舒悦李双宁保安
Owner INST OF ENVIRONMENTAL MEDICINE & OCCUPATIONAL MEDICINE ACAD OF MILITARY MEDICINE ACAD OF MILITARY SCI
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