Preparation and application of a nucleic acid aptamer-modified magnetic metal-organic framework medium
A nucleic acid aptamer and magnetic metal technology, which is applied in the field of material science and modern separation and analysis, can solve the problems of poor selectivity and solvent resistance, and achieve the effect of high extraction capacity and good selective enrichment ability
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Embodiment 1
[0029] The preparation of the magnetic MIL-101 of embodiment 1 nucleic acid aptamer modification
[0030] The synthetic schematic diagram of OTA Apt-MMIL-101 of the present invention is as follows figure 1 Shown:
[0031] (1) NH 2 -Fe 3 o 4 Preparation: Accurately weigh 28.1g FeCl 3 ·6H 2 O, 14.4g FeSO 4 ·7H 2O, dissolved in 150mL of distilled water, poured into a 500mL three-necked flask, stirred mechanically, and passed through nitrogen to remove oxygen for 10min. Quickly add 70 mL of 25% ammonia water dropwise, then drop 10 mL of oleic acid into the flask at a constant rate of 0.5 mL / min, and stir at constant temperature for 1 h. The obtained magnetic fluid was placed on a magnet to settle for 20 minutes, and then the supernatant liquid was poured off, and the magnetic separation was cleaned by ultrasonic cleaning with dedistilled water. Measure 15mL of magnetic fluid and place it in 200mL of n-propanol, after ultrasonication for 15min, add NH under mechanical stir...
Embodiment 2
[0035] The selectivity and enrichment ability of embodiment 2OTA Apt-MMIL-101
[0036] (1) Optional
[0037] OTA Apt-MMIL-101 has specific recognition performance for its target, select 6 analogues of ochratoxin A, ochratoxin B (OchratoxinB, OTB), aflatoxin B1, aflatoxin G1 , aflatoxin G2, acemethaquine and quinocetone. The adsorption capacity of OTA Apt-MMIL-101 to different compounds was evaluated by the absolute adsorption capacity of MSPE extraction concentration of 10 μg / L. Standard solutions of 10 μg / L of each substance were prepared with hydroxyethylpiperazine ethylsulfuric acid (HEPES) buffer solution, and the extraction volume was 10 mL. The results are shown in Table 1.
[0038] Table 1 Adsorption performance of OTA Apt-MMIL-101 on different substances
[0039]
[0040] As shown in Table 1, the OTA Apt-MMIL-101 prepared by the present invention has a very high selective extraction ability for ochratoxin A (OTA). However, for the structural analogues of OTA, su...
Embodiment 3
[0042] Example 3 Application of OTA Apt-MMIL-101 combined with HPLC-FLD to analyze trace amounts of ochratoxin in food
[0043] (1) Establishment of HPLC-FLD analysis method
[0044] Prepare a series of ochratoxin A standard solutions with concentration gradients of 20.0, 10.0, 2.00, 1.50, 1.00, 0.500, 0.100, 0.0500, 0.0250 μg / L, etc., and modify the magnetic MIL- The standard curve, linear range, correlation coefficient and detection limit of ochratoxin A determined by 101 solid phase extraction combined with UPLC-FLD. Within the set concentration range, the standard equation of ochratoxin A is Y=3.71×10 5 X+1.14×10 4 , the linear relationship at 0.025-20 μg / L is good (correlation coefficient is 0.9998), adopting 1 μg / L standard solution to investigate method precision (n=5) RSD is 3.2%, detection limit 6.7ng / L (detection limit according to Signal-to-noise ratio 3:1 estimated).
[0045] (2) Analysis of actual samples
[0046] Weigh 10.0g powder of crushed rice or peanut ...
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