Electrochemical immunosensor detection method for capsaicin substances and application
An immunosensor, capsaicin technology, applied in the direction of material electrochemical variables, scientific instruments, biological testing, etc., can solve the problems of long operation time and high test cost, and achieve the effect of enhancing stability and improving antibody immobilization effect
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Embodiment 1
[0029] Example 1: Preparation of immunosensor
[0030] (1) Preparation of nanomaterials
[0031] First, all glassware was treated with aqua regia and rinsed with ultrapure water before drying to ensure that there were no impurities on the glassware that might affect the experimental results; 50 mL of 0.01 wt% tetrachloroauric acid solution was heated under magnetic stirring to the boiling point, then quickly add 2.5 mL of 1wt% sodium citrate solution; when the mixture turns dark red, continue to boil and stir for 15 minutes, then cool to room temperature, and store in the dark at 4°C; weigh 0.1 g chitosan, dissolve In 200 mL of 1.0% acetic acid solution, stir at room temperature for 3 hours to completely dissolve chitosan; 0.0625g FeCl 3 , 0.0822g K 3 [Fe(CN) 6 ], 0.7455g KCl and 1 mL of concentrated hydrochloric acid were added to 100 mL of the above-mentioned chitosan solution, and dispersed at room temperature by ultrasonic waves to obtain a stable dark green dispersion;...
Embodiment 2
[0042] Embodiment 2: An electrochemical immunosensor based on thionine / Prussian blue-nano gold composite material is used for the detection of capsaicin in serum
[0043] (1) Establishment of standard curve
[0044] In order to monitor the electrochemical signal changes occurring on the immunosensor, the differential pulse voltammetry detection technique was used at 5.0 mM [Fe(CN) 6 ] 3− / 4− It was carried out in solution (pH 7.5, containing 0.1 M KCl), and the potential range was -0.1~+0.6V (potential increment 5 mV / s, amplitude 50 mV); all immunosensors were prepared under optimal experimental conditions, and the measurement Its initial DPV response (recorded as I); then, 5 μL of different concentrations of capsaicin standard was dropped on the immunosensor, incubated at room temperature for 40 minutes, rinsed and dried and then detected its electrochemical response again (recorded as I 0 ), calculate the ΔI value (ΔI=I−I 0 ), analyze its relationship with capsaicin standa...
Embodiment 3
[0047] Example 3: An electrochemical immunosensor performance test for detecting capsaicin in blood based on thionine / Prussian blue-gold nanocomposites
[0048] (1) Selective analysis
[0049] Figure 5 where abc stands for ferulic acid, isoferulic acid and trans-ferulic acid respectively, d is the mixture of these three interfering substances, efg is the three specific targets of the sensor capsaicin, synthetic capsaicin and dihydrocapsaicin , h is the mixture of these three capsaicinoids, and i is the mixture of these six substances; it can be seen from the figure that when there is no specific target substance, the DPV peak current difference is very small, indicating that the antibody cannot interact with These interfering molecules are combined; the histograms h and i have a large current difference, and the results are similar, indicating that the immunosensor has good selectivity and anti-interference ability.
[0050] (2) Stability and repeatability studies
[0051]...
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