Method for detecting trace silver ions based on biosensor
A biosensor and silver ion technology, applied in the field of bioelectrochemical sensors, can solve the problems of cumbersome preparation and detection, large background noise, and high cost, and achieve the effects of low background noise, simple equipment, and wide linear range
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Embodiment 1
[0047] Construction of ECL biosensors.
[0048] The schematic diagram of the construction of this ECL biosensor is attached figure 1 shown, including the following steps:
[0049] (1) Synthesis of citrate AuNPs and linkDNA-AuNPs.
[0050] Synthesis of 1’ citrate AuNPs: Add 98 mL of deionized water into a double-necked flask, and add HAuCl4 to make the final concentration 1 mM. When the solution started to reflux, sodium citrate solution (10 mL, 38.8 mM) was added quickly and the stopper was closed. The color of the solution changed from light yellow to deep red after 1 minute, and the reflux was continued for 20 minutes. Turn off the heat and allow the system to cool to room temperature (23-25 °C) with stirring. The color of the prepared nanoparticle solution should be deep red, with high stability and reproducibility, and it can be stable for several months when stored in a clean container. As shown in the transmission electron microscope picture (attached figure 2 ...
Embodiment 2
[0058] The ESI method verified the successful construction of the ECL biosensor.
[0059] Bare GE has a smaller signal (attached image 3 A, Curve a), and when the probe DNA is immobilized on the GE surface and blocked with MCH, the ECL signal intensity decreases slightly due to the hindrance of the nucleic acid to the electron transfer (attached image 3 A, curve b). When linkDNA-AuNPs were connected to the electrode surface through hybridization with probe DNA, the intensity of ECL increased, which proved that the strong electron transfer ability of AuNPs can promote the reaction of ECL (attached image 3 A, curve c). When Ag is added + After that, the ECL signal intensity was significantly enhanced, which is due to the large number of AuNPs connected to the surface of the electrode and the C-Ag + The formation of -C structure greatly improves the charge transfer rate (attached image 3 A, curve d). Bare GE exhibits a smaller semicircle (attached image 3 B, Curv...
Embodiment 4
[0061] Differential pulse voltammetry to verify ECL signal and C-Ag + -C relationship.
[0062] attached Figure 4 A compared the biosensors constructed by AuNPs modified with DNA sequences rich in four different bases connected to the electrode surface against Ag + Link DNA 1, Link DNA 2, Link DNA 3 and Link DNA 4 respectively correspond to DNA sequences rich in C, T, A, and G bases.
[0063] Depend on Figure 4 In A, we can clearly see that Ag +Sensors constructed only to C-base-rich DNA sequences responded, demonstrating a significant increase in ECL signal with C-Ag + The formation of -C has an indispensable relationship. as attached Figure 4 As shown in B, we tried to use M DNA instead of probe DNA to carry out control experiments. M DNA is a single-stranded DNA rich in A bases and cannot be complementary to link DNA. attached Figure 4 (a) in B represents the pair of sensors constructed by hybridizing AuNPs modified by Link DNA 1 and Link DNA 2 to probe DNA, ...
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