Theses (MSc in Analytical and Pharmaceutical Chemistry)

Tocotrienols (T3), a subfamily of vitamin E, are gaining recognition for their potent antioxidant, anti-inflammatory, and neuroprotective properties. However, their use in functional foods and medicinal products faces challenges such as low water solubility, instability under environmental conditions, and susceptibility to oxidative degradation. This study aims to address these challenges by using spray drying as an encapsulation technique to convert liquid nano emulsions of T3 into stable, dry powders. The process is guided by the Quality by Design (QbD) methodology, which optimises the spray drying parameters and the selection of wall materials to enhance the stability of nano-T3 powders. The results indicate that the combination of sodium caseinate and fibersol as wall materials effectively encapsulated T3, preserving its chemical integrity during processing. The optimised spray drying conditions produced nano-sized particles with ideal characteristics such as excellent flowability (10.71 ± 1.12%), wettability (93.07 ± 6.01%), low moisture content (1.79 ± 0.02%), particle size (100.42 ± 48.05 nm) and Vitamin E content (579.00 ± 233.39 mg/g). However, stability testing revealed that while the nano-T3 powder retained its chemical stability, moisture absorption over time led to particle aggregation and a decrease in T3 content. This indicates a need for further optimisation to improve moisture resistance and long-term stability. The study highlights the significant impact of wall materials and spray-drying parameters on the properties of nano-T3 powders and provides valuable insights into enhancing their stability and efficacy. Future research should focus on refining the encapsulation matrix and exploring the incorporation of antioxidants to mitigate degradation, thereby improving the stability and functional applications of nano-T3 in pharmaceuticals, nutraceuticals, and functional foods. In conclusion, this study demonstrates the potential of spray-dried nano-T3 as a stable, bioactive ingredient for diverse applications, with room for improvement in moisture resistance to enhance long-term stability.

The emergence and re-emergence of viral pathogens such as SARS-CoV-2, human immunodeficiency virus (HIV-1), hepatitis C virus (HCV), Zika virus (ZIKV), and dengue virus (DENV) continue to pose significant global health challenges. Viral proteases play a central role in viral replication through polyprotein processing and therefore represent attractive targets for antiviral drug development. This dissertation investigates the identification of synthetic viral protease inhibitors using an integrated in silico approach comprising Structure-Based Pharmacophore Modelling (SBPM), virtual screening, molecular docking, and pharmacokinetic prediction. High-resolution crystal structures of five disease-specific viral proteases (PDB IDs: 8HUR, 4WF8, 3SPK, 2FOM, and 5H4I) were analyzed to generate validated structure-based pharmacophore models using the Pharmit platform. These models were applied to virtually screen large compound libraries, including ZINC and MolPort, to identify molecules exhibiting key interaction features within protease active sites. The top-ranked virtual hits were further evaluated through molecular docking simulations using iGEMDOCK to predict binding conformations, interaction profiles, and relative binding affinities. Drug-likeness and ADMET properties were assessed using the SwissADME web tool. Several chemically diverse lead scaffolds, including triazine, indazole, and isoquinoline cores, demonstrated favorable hydrogen bonding, π–π stacking, and hydrophobic interactions with conserved catalytic residues across multiple viral proteases. Notably, some compounds exhibited multi-target binding potential, indicating suitability for broad-spectrum antiviral development. Overall, this study demonstrates that SBPM integrated with molecular docking provides an efficient and cost-effective computational framework for antiviral lead identification and establishes a strong foundation for future experimental validation and optimization. Keywords: viral proteases; structure-based drug design; pharmacophore modelling; virtual screening

Wound healing is a complex biological process requiring a coordinated response to restore damaged tissue. Traditional wound dressings such as gauze and bandages provide protection but lack bioactivity, delaying healing. In addition, drug-resistant bacteria are an ever-evolving issue that poses a problem. This limitation can lead to prolonged recovery times and increased susceptibility to infections. This study investigated sodium alginate-carboxymethyl cellulose composite hydrogels incorporating bioactive glass and alizarin for wound healing applications. Formulations made up of sodium alginate and sodium carboxymethyl cellulose, alizarin, and bioactive glass were prepared via lyophilisation. Fourier transform infrared spectroscopy confirmed the presence of sodium alginate, sodium carboxymethyl cellulose, alizarin, and bioactive glass and indicated successful incorporation of these components. Scanning electron microscopy explored the porosity of the samples and proved that alginate-carboxymethyl cellulose hydrogels have the most pores present to facilitate and house active components. Antioxidant assays showed alizarin’s dose-dependent radical scavenging activity. At the same time, drug-release studies indicated near-complete alizarin release within 168 hours. Degradation in PBS was slower in bioactive glass-rich formulations. Antibacterial studies against Staphylococcus aureus and Pseudomonas aeruginosa showed that samples with an equal ratio of bioactive glass and alizarin showed the highest reduction in the bacteria strains compared to the control. MTT assays indicated that alginate-carboxymethyl cellulose hydrogels and samples with bioactive glass only exhibited the highest number of living and healthy cells compared to alginate-carboxymethyl cellulose with alizarin incorporated. Finally, in scratch assays, the sample with equal alizarin and bioactive glass ratios displayed the most rapid wound closure, suggesting that a balance of components can significantly enhance wound healing. Overall, this study demonstrates that combining sodium alginate, sodium carboxymethyl cellulose, bioactive glass, and alizarin can produce hydrogels with promising antioxidant, antibacterial, and tissue-regenerative properties for wound-healing applications. However, further optimisation may be required to balance cytocompatibility and antibacterial efficacy.

Nanofibers have become a prominent research focus in wound dressing applications due to their exceptional material properties. The drug-loaded nanofibers can effectively eliminate the bacterial growth in the wound site. This study presents the formulation and fabrication of chitosan-gelatin (CS-GEL) nanofibers incorporated with curcumin and copper/zinc-based metal-organic framework (Cu/Zn-MOF) using the electrospinning method. The Cu/Zn-MOF was analysed using X-ray diffraction (XRD). The CS-GEL nanofibers loaded with curcumin and copper/zinc MOF were characterized with ATR-FTIR spectroscopy and scanning electron microscopy (SEM). The in-vitro antioxidant, anti-inflammatory, cytotoxicity, and antibacterial properties were also evaluated. The optimal conditions for smooth nanofiber production were identified as a polymer concentration of 3% (w/v) chitosan and 45% (w/v) gelatin, with a 30:70 chitosan-to-gelatin ratio. The optimized electrospinning parameters included an applied voltage of 18 kV, a flow rate of 0.8 mL/h, and a working distance of 15 cm. The optimized concentration of curcumin-Cu/Zn-MOF/CS-GEL nanofibers showed no toxic effects on HaCaT cells in 5% (w/v) of curcumin. Additionally, the 5CUR/10CuZn-MOF/CS-GEL nanofiber sample showed the highest percentage of DPPH radical scavenging (83.70%) for antioxidant activity. The highest percentage inhibition for the formulation of curcumin and MOF was 83.10%. The incorporation of curcumin and Cu/Zn-MOF effectively inhibited the growth of S. aureus and P. aeruginosa in antibacterial assays. The 5CUR/10CuZn-MOF/CS-GEL nanofiber sample was the most effective formulation against S. aureus and P. aeruginosa for antibacterial test. The 5CUR/10CuZn-MOF/CS-GEL nanofiber sample also showed the anti-biofilm effect against S. aureus. The 5CUR/1CuZn-MOF/CS-GEL, 5CUR/5CuZn-MOF/CS-GEL and 5CUR/10CuZn-MOF/CS-GEL nanofiber samples demonstrated potential applications for wound healing, as they were non-toxic to cells and exhibited antibacterial activity, which showed wound healing rate of 95.30%, 99.30% and 96.96%, respectively. The potential application of advanced wound dressings and nanotechnology lies in their ability to enhance healing rates and improve treatment outcomes.

Omega-3 fatty acids (ω-3 FAs) includes eicosapentaenoic acid (EPA) and docosahexaenoic acid (DHA) have attracted considerable interest for their likely health advantages. Many different benefits have been claimed for ω-3 fish oils, including preventing cardiovascular diseases (CVD), reducing cognitive decline, and helping improve and manage inflammatory diseases such as asthma, neurodegeneration, and arthritis. This study investigates the chemical profiles and heterogeneity of various commercial omega-3 fish oil products by chemometric cluster analysis of spectroscopic data, with specific focus on correlating these findings with their neuroprotective activity in microglial cells. Acquiring spectra was enabled using spectrophotometers: FTIR (Shimadzu) and 1H-NMR-Nanalysis equipped with MestReNova software operating at 61 MHz. A bioassay experiment was performed on the BV2 Murine microglia cell line, where Nitrite (NO) production was measured through the Griess assay. The NMR spectra were analyzed through chemometric analysis by principal component analysis (PCA), identifying patterns for the classification of the samples by determining similarities among them based on potency and establishing the relationship between the spectra data and NO production of the samples. The assessment of fish oils using spectroscopy techniques (FTIR and 1H-NMR) revealed the presence of ω-3 FAs compounds as their spectra fitted the DHA and EPA reference standards. The concentrations of the fish oils corresponded relatively inversely with the NO production measured through the Griess assay. DHA and EPA significantly reduced NO production on BV2 microglial cells, revealing the neuroprotective properties. The concentration range of DHA and EPA determined in samples was meaningful to exhibit the corresponding neuroprotective properties of the compounds.