Method for detecting norfloxacin in milk based on surface enhanced Raman technology
A surface-enhanced Raman and technical detection technology, which is applied in the field of analysis and detection, can solve the problems of complex experimental process, low detection limit, and low detection limit, and achieve the effect of improving capture ability, high accuracy, and simple and fast detection
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Embodiment 1
[0050] The preparation of embodiment 1 substrate
[0051] β-cyclodextrin is used as a reducing agent to reduce silver nitrate to obtain β-cyclodextrin-modified silver nanoparticles as a method for preparing a Raman-enhanced substrate, comprising the following steps:
[0052] (1) Weigh 0.0851g of β-cyclodextrin and dissolve it in 50mL of deionized water, keep stirring at 500rpm for 10 minutes to obtain the β-cyclodextrin solution;
[0053] (2) Take 30mL of β-cyclodextrin solution and add it to a conical flask, heat it in a water bath at 60°C for 10 minutes, add 15mL of deionized water, and add the same volume of 7.5mM sodium hydroxide solution to adjust the pH to 10-12 to obtain a base Stirring environment; the solution was kept in a water bath at 60°C and stirred vigorously at 700rpm for 5 minutes;
[0054] (3) Slowly add 20 mL of silver nitrate solution with a concentration of 1 mM in the solution of step (2), and the speed control is to add 200 μL of silver nitrate solution...
Embodiment 2
[0055] The construction of embodiment 2 standard curve model
[0056] (1) Configure 10 first -3 The norfloxacin mother liquor of M concentration, gets mother liquor and is diluted to the norfloxacin standard solution of 0.025, 0.05, 0.125, 0.25, 0.375, 0.5, 2.5 and 5 μ mol / L respectively afterwards;
[0057] (2) Norfloxacin standard solutions of different concentrations and milk are mixed evenly according to the volume ratio of 1:1 to form milk sample solutions of different concentrations of norfloxacin;
[0058] (3) Milk sample liquids of different concentrations and acetonitrile were mixed uniformly according to the volume ratio of 1:1, and centrifuged at 12000rpm for 10min to obtain supernatants of different concentrations of norfloxacin;
[0059] (4) Mix the supernatants of different concentrations with the SERS substrate according to the volume ratio of 1:1, let it stand for 10 minutes, and perform Raman detection to obtain the Raman spectra of milk sample liquids with d...
Embodiment 3
[0067] A method based on surface-enhanced Raman technology detection of norfloxacin in milk (flow process as follows Figure 4 ), including the following steps:
[0068] (1) Mix milk with norfloxacin concentrations of 0.375, 0.25, and 0.05 μmol / L with acetonitrile at a volume ratio of 1:1, and centrifuge at 12,000 rpm for 10 minutes to obtain a supernatant;
[0069] (2) Mix the supernatant and the β-cyclodextrin-modified silver nanoparticle substrate (β-cyclodextrin-AgNPs) prepared in Example 1 evenly according to the volume ratio of 1:1, let stand for 10min, and perform Raman Detect, obtain the Raman spectrogram of milk (such as Figure 5 );
[0070] (3) 1389cm in the Raman spectrum -1 ±5cm -1 Characteristic peak intensity is substituted into the standard curve model of embodiment 2, obtains the concentration of norfloxacin in the milk to be tested.
[0071] The test results are as follows:
[0072] The milk sample to be tested with a norfloxacin concentration of 0.375 ...
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