Colloidal gold test strip for detecting sildenafil drugs, and preparation method and application thereof
A technology for sildenafil and test strips, which is applied in the field of colloidal gold test strips for sildenafil drugs and its preparation, can solve problems such as low sensitivity, inability to realize multi-residue immunoassay detection, and few detection objects
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Embodiment 1
[0084] The preparation of embodiment 1 sildenafil drug colloidal gold test strip
[0085] 1) Preparation of coating original
[0086] Utilize active ester method to prepare coating former, concrete steps are as follows:
[0087] (1) get the sildenafil hapten 13.2mg (0.05mmol) shown in formula (I), be dissolved in 1.5mL DMF, stir and add 24.8mg DCC and 13.8mg NHS, stir and dissolve and prepare C liquid; Dissolve 30 mg of ovalbumin in 4 mL of PBS buffer (0.01M, pH 7.4), stir and dissolve to prepare solution D; under magnetic stirring, absorb solution C and add it to solution D dropwise, and react with magnetic stirring at 4°C for 12 hours; centrifuge Finally, take the supernatant, dialyze against PBS buffer solution at 4°C for 3 days, change the dialysate 3 times a day, and obtain the coating agent with the structural formula as shown in formula (VII);
[0088] (2) Adjust the concentration of PBS buffer used for coating prepared in step (1) to 1 mg / mL, dispense 500 μL of each ...
Embodiment 2
[0117] The using method of embodiment 2 test strips
[0118] The using method of the test strip that above embodiment 1 obtains may further comprise the steps:
[0119] 1) Sample pretreatment
[0120] (1) Wine sample processing
[0121] Take 1.0mL alcohol sample in a centrifuge tube, dry it with nitrogen in a water bath at 50°C, redissolve with 50μL 0.1% acetic acid-methanol solution, then dilute to 1.0mL with 0.1M PB buffer, vortex for 30s, and set aside;
[0122] (2) Oral liquid sample processing
[0123] Take 0.2mL of liquid sample in 5mL of chloroform organic solvent, adjust the pH of the solution to 9.0 with ammonia water, vortex and shake for 1min, and let it stand to separate layers; take the organic reagent part, dry it with nitrogen in a water bath at 50°C, and use 50μL of 0.1% Reconstitute with acetic acid-methanol solution, then dilute to 0.2mL with 0.1M PB buffer, vortex for 30s, and set aside;
[0124] (3) Solid sample processing
[0125] Take 0.5g of solid s...
Embodiment 3
[0136] Sensitivity and specificity detection of embodiment 3 test strips
[0137] 1. Sensitivity and specificity detection experiments of the kit
[0138] The colloidal gold test strips obtained in Example 1 of the present invention were used to detect sildenafil drugs (as shown in Table 1), and each drug was prepared in different concentrations.
[0139] 2. Sensitivity and specificity test results
[0140] The test strip that the embodiment of the present invention 1 obtains is to the detection sensitivity of sildenafil drugs, specificity and standard curve as table 1 and figure 2 As shown, the results show that the test strip is effective for Sildenafil, NorneoSildenafil, N-Desmethyl Sildenafil, Homocidena HomoSildenafil, Thiohomosildenafil, Aildenafil, Thioaildenafil, Prepoxypheyl Thioaildenafil , Acetildenafil, Hydroxyacetildenafil, Udenafil, Vardenafil, Pseudo Vardenafil, or Namovardenafil (NorneoVardenafil) The detection results of 14 sildenafil drugs were all posit...
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