The FTIR analysis reveals that crystals resulting from the MA-MAEC technique are GSH-based. “GSH was isolated in yeast for the first time in 1929, and its metabolism in the body and its role in cancer treatment was described in 1984” (Natter and Kohlwein 315; Ortega, Mena, and Estrela 1286). MA-MAEC is a technique that merges silver nanoparticle films (SNFs) and microwave heating in producing temperature gradients induced by microwaves, which boost the formation and growth of crystals.
Crystallization of Glutathione by using cenvenatil microwave
Metal-assisted and microwave-accelerated evaporative crystallization (MA-MAEC technique) was used to ascertain the duration of crystallization of both substrates, namely, PMMA substrate and PMMA substrate coated with SNFs, at room temperature. The duration of crystallization was the time taken for the initial crystals to form and the length of time it took for complete crystals to form. The outcomes of all GSH concentrations, which were 500, 400, and 300 mg/ml, are summarized in Table 2 and 3. Comparatively, the duration of crystallization at microwave and room temperature conditions for the SNFs substrate was 25-30% faster than for the PMMA substrate. The experimental outcomes show that 300 mg/ml of GSH in PMMA platform took an average duration of 628±314 minutes. In comparison, the experimental outcomes also show that 300 mg/ml of GSH in SNFs coated PMMA platform took 415±38 minutes. As Table 3 shows, the durations of crystallization for SNFS coated platform and PMMA platform were 108±12 minutes and 130±20 minutes correspondingly.
The analysis of the experimental outcomes reveals that microwave heating plays a central role in the MA-MAEC process because it speeds up the rate of crystallization. Evidently, the rate of crystallization under microwave conditions is 5.8 times faster than the rate of crystallization under the room temperature conditions.
The GSH crystals produced at room temperature through the MA-MAEC technique in both SNFs coated platform and PMMA platform have matching diffraction peaks, which resemble the X-ray diffraction peaks indicated in the literature. The matching and resemblance of diffraction peaks indicate that MA-MAEC process does not change the structure and form of GSH crystals.
- Ethyl alcohol, which was bought from Pharmaco-Aaper in Brookfield
- EMS 150RS silver targets and a sputter coater, which was bought from Electron Microscopy Sciences in Hatfield
- 0.01 g/ml solution of GSH was prepared by weighing 0.01 g and putting it in 1 ml solvent. To dissolve GSH completely, the solute and the solvent were shaken vigorously for five minutes. To enhance dissolution of GSH at room temperature, a Corning magnetic stirrer was used. However, 1ml of DMF and ethanol did not dissolve 0.01 g of GSH to make a concentration of 0.01 g/ml despite vigorous shaking for 60 minutes. GSH was weighed and gradual increments of 0.01 g were added to the solvents while shaking vigorously by means of a Corning stirrer pending complete dissolution of GSH. The visual appearance of the solute was used to judge the extent of dissolution and the temperature as determined using thermometer.
Preparation of Sodium acetate buffer Solution (pH=4.6).
To prepare 0.2 M solution of sodium acetate, 8.2 g of the salt was dissolved in 500 ml of deionized water. Hydrochloric acid and/or sodium hydroxide were added appropriately to the solution to adjust the pH to 4.6. In the analysis of the crystals formed, the 2.0 version of Motic images software was used. Moreover, FTIT spectrometer was used to generate infrared spectrum of GSH crystals.
Comparison of crystallization time between iCrystal system and commercial microwave for Glutathione
Fourier Transform Infrared Spectroscopy (FTIR) for GSH
The MA-MAEC technique employed in all concentrations of PMMA and PMMA coated SNFs, as shown in Figures 25-35, illustrates the infrared spectra produced by Glutathione crystals subjected to microwave heating at 200-1000 watts. The infrared spectral peaks for Glutathione were CN stretch at 1075 cm-1, COO– symmetric stretching at 1382 cm-1, NH stretching at 334 and 3246 cm-1, C=O anti-symmetric at 1710 cm-1, and SH stretch at 2520 cm-1.
Subsequently, the MA-MAEC technique was employed in the successful synthesis of Glutathione. The analysis of the synthesized GSH crystals using FTIR revealed that the functional groups were intact and undisturbed by the MA-MAEC process. The most significant functional groups of GSH that were present are the –CN stretch, amine (-NH2/-NH), anti-symmetric (C=O), carbonyl group, thiol group (-SH), and asymmetric group (COO–).
Natter, Klaus, and Sepp Kohlwein. “Yeast and cancer cells: Common principles in lipid metabolism.” Biochimica et Biophysica Acta 1831.1 (2013); 314-326. Print.
Ortega, Anger, Salvador Mena, and Jose Estrela. “Glutathione in Cancer Cell Death.” Cancers 3.1 (2011): 1285-1210. Print.