Preparation method of polyglutamic acid PGA-coated superparamagnetic iron oxide nanoparticle
A technology of iron oxide nanometer and polyglutamic acid, which is applied in the preparations and pharmaceutical formulations for in vivo experiments, can solve the problem of not finding SPIO nanoparticles, and achieve a remarkable contrast effect, good water solubility, and low raw material cost. Effect
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[0061] Example 1
[0062] Dissolve 640 mg of ferric chloride hexahydrate in 50 mL of ultrapure water, bubbling in nitrogen for 10 minutes and fully stirring. Dissolve 90 mg of PGA in 20 mL of ultrapure water, and add the above solution dropwise. After stirring well, add 9mg / mL sodium sulfite solution dropwise. Transfer the mixed solution to a 60°C water bath, add 1 mL of ammonia water, and fully react for 30 minutes. The mixed solution was moved to room temperature, and the reaction was continued for 1.5 hours. Centrifuge the prepared solution, centrifuge at 8000 rpm for 10 minutes, discard the centrifugal sediment, and take the upper layer solution. Dialysis with a dialysis bag with a molecular weight cut-off of 8000-14000. Distilled water was used for dialysis, and the water was changed three times a day after dialysis for three days. The polyglutamic acid-coated ferroferric oxide nanoparticles prepared by the present invention were observed through a transmission electron...
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[0063] Example 2
[0064] The concentration of Fe in the SPIO-PGA nanoparticle solution prepared by the present invention is measured by the ICP-OES test method. Prepare 2 mL of SPIO-PGA nanoparticle aqueous solution with Fe concentrations of 0.004, 0.008, 0.016, 0.032, 0.064mM, and measure the T of the material under different Fe concentrations by magnetic resonance imaging analyzer. 2 Relaxation effect (see attached Figure 5 ). The relaxation rate test results show that the reciprocal relaxation time of SPIO nanomaterials has a good linear relationship with the increase of iron concentration (within the range of 0.004-0.064mM concentration). The r of SPIO-PGA prepared by the present invention can be obtained by calculation 2 Relaxation rate up to 333.7mM -1 s -1 . Therefore, the SPIO-PGA prepared by the present invention can be used as an excellent T 2 Signal attenuation contrast agent.
Example Embodiment
[0065] Example 3
[0066] HeLa cells are used as model cells to evaluate the effect of SPIO-PGA nanoparticles prepared by the invention on cell viability. Naked Fe prepared in Comparative Example 1 3 O 4 Nanoparticles served as a control. The solution obtained in Example 1 was sequentially prepared with Fe element concentration of 50, 150, 250, 350, 450 μg / mL SPIO-PGA nanoparticles physiological saline solution (sodium chloride content 0.9%). Plant HeLa cells in a 96-well plate, set 5 parallel samples for each concentration, and place the cell culture plate in CO 2 The concentration was 5% and the temperature was 37°C for 24 hours. After MTT treatment, the absorbance value of each well at λ=570nm was detected on a microplate reader, and the corresponding cell viability was calculated based on this. The cells treated with saline were used as a blank control, and the cell viability was recorded as 100%. Naked Fe with control group 3 O 4 In comparison, SPIO-PGA-treated cells have ...
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