Method and kit for detecting atrazine, chlorpyrifos and triadimefon based on SERS technology
A detection kit and atrazine technology, applied in the field of pesticide detection, can solve the problems of low detection sensitivity, achieve high stability, and overcome the effect of long preparation cycle
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[0038] In some embodiments, the preparation method of the artificial antigen of atrazine, chlorpyrifos, triadimefon comprises:
[0039] The hapten of the pesticide molecule is activated and reacted with NHS and EDC in turn, and then coupled with the carrier protein;
[0040] The pesticide molecule hapten is selected from any one of atrazine, chlorpyrifos and triadimefon.
[0041] In some embodiments, the molar ratio of the pesticide molecule hapten to NHS and EDC is 1: (1.7-2.3): (2.7-3.3);
[0042] In some embodiments, the molar ratio of the pesticide molecule hapten to NHS and EDC is 1:2:3.
[0043] In some embodiments, the carrier protein is BSA.
[0044] According to another aspect of the present invention, the present invention also relates to a method for detecting pesticides using the above kit, the method is used to detect one or more of atrazine, chlorpyrifos, triadimefon, the method include:
[0045] The substance to be detected is used to competitively bind to t...
Embodiment 1
[0048] 1. Synthesis of nano gold sol
[0049] First heat 300mL 0.01% chloroauric acid solution to boiling, quickly add a certain amount of 1% trisodium citrate aqueous solution, and obtain nano-gold sols with different particle sizes by changing the amount of sodium citrate added. Some blue at first, then light blue, then blue, then heated to red, boiled for 7 to 10 minutes to appear transparent orange-red, wherein the molar ratio of trisodium citrate to the chloroauric acid is 4.3.
[0050] 2. Preparation of signal-labeled probes based on gold nanospheres (AuNPs) / carrier protein / target
[0051] Triazophos hapten carrier protein, atrazine hapten carrier protein and chlorpyrifos hapten carrier protein were respectively selected, and covalent bond, electrostatic self-assembly and other reconstruction techniques were used to prepare gold nanospheres (AuNPs) / carrier protein / Three different signal-labeled probes composed of targets.
[0052] (1) Triazophos hapten (15mmol L -1 ,...
Embodiment 2
[0062] 1. Synthesis of nano gold sol
[0063] First heat 300mL 0.01% chloroauric acid solution to boiling, quickly add a certain amount of 1% trisodium citrate aqueous solution, and obtain nano-gold sols with different particle sizes by changing the amount of sodium citrate added. Some blue at first, then light blue, then blue, red after heating, and transparent orange-red after boiling for 7-10 minutes. In the experiment, it was found that when the molar ratio of trisodium citrate: chloroauric acid was 4.9, it was possible to synthesize gold nanoparticles of about 16nm. Stable storage for more than half a year.
[0064] 2. Preparation of signal-labeled probes based on gold nanospheres (AuNPs) / carrier protein / target
[0065] Triazophos hapten carrier protein, atrazine hapten carrier protein and chlorpyrifos hapten carrier protein were respectively selected, and covalent bond, electrostatic self-assembly and other reconstruction techniques were used to prepare gold nanospheres...
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