Results of protein and MTX loaded onto magnetic nanoparticlesFe3O4@ (SiO2/SHor SiO2/SH/NH2) by spectrophotometry
The purpose of this section is to investigate the absorption of drug biomolecules on the magnetic nanoparticle bed. Therefore, using the equation [7-8], we can examine the absorption rate. Under standard conditions, the amount of 30 micrograms per μl of the bio-molecule protein and drug are dissolved in 2 ml of sterile water, and then twice 25 milligrams of magnetic nanoparticles weigh in two separate dishes and on each One of the bio-molecule proteins and drug (60min, 36h) were solved separately.
The instant of dissolution of the two mixtures was continued at the instant of zero minutes to about an hour for each of the mixtures and summarized the obtained data. The results showed that the absorption rate of the protein in the first half hour was about 60% and the absorption rate of the drug on the nanoparticles were about 80%. The results obtained in the future did not change later, and the results showed that the absorption of nanoparticles in non-co-volcanic and covalent magnetic nanoparticles is approximately equal to one. After methotrexate has been stabilized on a surface of Fe3O4@SiO2/SH/NH2 magnetic nanoparticles with different methods, we will now consider the methotrexate release rate. This is done with a phosphate buffer solution which is a mixture of 4 K2HPO3, Na2HPO3, KCl, and NaCl salts in a specified amount of distilled water, which is ready for use in the reaction after the autoclave. Using phosphate buffered saline, sodium hydroxide, and chloride acid adjust the pH to 8.8 to adjust the pH of the pH in the presence of phosphate buffer of methotrexate on the surface of nanoparticles and with sodium hydroxide and chloride. And the rate of release has been measured over a period of 2 hr to 72 hr at pH of 7.28-8.0. Looking at the data in the chart, it can be concluded that after 24 hr, the release of the methotrexate drug molecule from the surface of nanoparticles is about 80% at pH 7.8. With these results, can be said that is a full stabilization of MTX drug molecule on MNPs-IHSPN In-vitro. The results are shown in (Figure 3). All Error bars are selected as standard, indicating the accuracy of the data presented in the chart.
The effect of time in absorption
Reaction time for absorption, 1hr and 36hr in this test as the standard time for this research is very important. At the beginning of the reaction, patterns were to study the magnetic nanoparticles containing silica-protein-MTX absorption studies conducted showed that the uptake protein the concentration (30 μg/ml) and 10µg/ml for MTX, in The wavelength of 595nm for 30min (absorption wavelength protein) is 60% and the uptake MTX the concentration (10 μg/ml), in The wavelength of 304nm for 36h (absorption wavelength MTX) is 80%, and after 1hr and 36hr absorption be stable over time because the absorption process of protein and MTX was complete on the surface of magnetic nanoparticles. The absorption of biomolecules (MTX, protein) within a half-hour and twenty four-hour episodes were tested and the results were observed in (Figure 4). And according to the results, we find that over time gradually increased uptake, and it been stable after 1h for protein and 36h for MTX drug molecule.
The stabilization of methotrexate and protein to the surface of the magnetic nanoparticles Fe3O4@SiO2/SH/NH2 by EDX, FT-IR
In this discussion, by W% of elements perceived whichever were dependent to the reactants of MTX and protein and magnetic nanoparticles Fe3O4@SiO2/SH, Fe3O4@SiO2/SH/NH2 and EDX analysis showed that both of the reactants bonded together been in the product. Also, elements of Fekα and Fekβ with elements Si (with a strong peak) and O are shown in Fe3O4@SiO2 product. This analysis may be demonstrative bond between magnetic nanoparticles and MTX or BSA. The results of the EDX analysis show that binding of agent N in 750 keV and agent of O 1100 keV and agent N in 750 keV and agent of C of 600 keV because they are in a line so, it may be stated this approaching is electrostatic bonding same for N-O and covalent bonding same for C-N. Element O is in the agent group O2 of coated silica (SiO2) and element N of the agent NH2 of MTX or protein. Evidence of EDX analyses is a Spectrophotometer seconder for this tissue. The result of absorption, and the link between magnetic nanoparticles and MTX (down picture (B)) or BSA (up picture (A)) protein by EDX analysis, shown in (Figure 5, left picture).
In the FT-IR spectrum (Figure 5, right picture), by factorizing the nanoparticles with silica coating, we observe no change in the structure of the nanocomposite of magnetism. By examining the pixels of the four samples, the following results were obtained: A) the structure of magnetic nanoparticles with coating silica, B) the structure of magnetic nanoparticles with amine linker consisting of 2930 cm-1 ppm for the (-CH) group, 1407 and 1550 cm-1, for the -NH2 group and 1100 cm-1 for the Si-O group and furthermore so, 1112 cm-1 is for –SH group, C) Magnetic nanoparticle structure with SH-NH2 linker and fixed methotrexate on it includes 1606 and 1644 pixels cm-1 for covalent bond between carboxyl group (-COOH) methotrexate and amine group (-NH2), and D) methotrexate structure including peak 3391 cm-1 related the group (-OH) and peaks of and 1603 and 1644 cm-1 correspond to poor absorption of methotrexate on surface of an amine-bonded magnetic nanoparticle. -SH linker present in the ionic complex in the structure of the magnetic nanoparticles improves the covalent bond between the amine linker of the nanoparticles with the carboxyl linker of the methotrexate bio-drug and is a sharp and regular peak that shows drug absorption and strong bonding with the nanoparticles. So it can be said that the presence of ionic liquid composition increases the rate of drug absorption on the surface of the nanoparticles.
Stability of magnetic nanoparticles in repeated use after recycling
The results of the magnetic nanoparticles with silica coating for MTX or BSA adsorptions were analyzed by spectrophotometric analysis over a period of 12-170 h for 10 periods for protein and 4 days for MTX, and the results showed that the efficiency of nanoparticles in the application again and again the stabilization of biomolecules, MNPs-MTX, BSA even decreased by 10 percent over the course of 15 percent. Magnetic nanoparticles are very important for sustainability under favorable reaction conditions and having the ability to re-use these magnetic nanoparticles. On the picture 6, the magnitude of this stability has been investigated in 7 days (the reaction process and optimal conditions are the same as in the discussion and conclusion). The results are shown in (Figure 6).
The results of the absorption of protein by electrophoresis
In this section, protein is analyzed by the electrophoresis. Electrophoresis analysis is based on absorption at absorption times. Here, vertical electrophoresis to measure protein absorption. With respect to (Figure 7), it can be seen that the amount of stained specimens in the range of 0 to 60 mins, the absorption of protein on the magnetic nanoparticles have gradually dimmed, which this fading stain shows that biomolecules are absorbed on surface of magnetic nanoparticles. Therefore, spectrophotometric absorption and electrophoresis of both devices showed acceptable results for this absorption. First, in 5 different patterns for protein in three separate tests at a dose of 30 μg/ml and at 0-60min on a sol-gel plate (Different mixed supernatant solution and biomolecules (protein)-MNPs-IHSPN, and eventually the last point of the mass of magnetic nanoparticles containing biomolecules). In the first line, the ladder is first point, the second line of staining for protein and without nanoparticles (sample 1), the third line contains any protein mixed in nanoparticles (without catalytic) at zero time (sample 2), the fourth line contains either fixed protein in nanoparticles at 5 minutes (sample 3), fifth line at 15 minutes for protein (sample 4) and line six for 30 minutes for protein (Example 5). Then, sol-gel was placed on the electrophoresis and began to scan and stained. After 2-5 hours, results in same spots of about 60% of the apparently weaker spots were observed separately from the specimens containing the pure amount of the protein, and the staining smoothed showed that the absorption rate the nanoparticle surface is at least 60% for protein. Biomolecules protein is absorbed on surface of the magnetic nanoparticles, which confirms the stability of the biomolecules (protein) in the nanoparticles, which are shown in (Table 1 (down picture), Figure 7 (up picture)) evidence for this stability.