Dheyab, Ali Sami (2023) Extraction of anti-tuberculosis properties from selected medicinal plant by deep eutectic solvents (DES). Doctoral thesis, Universiti Tun Hussein Onn Malaysia.

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Abstract

Globally, the threat of tuberculosis (TB) continues to grow, requiring innovative techniques to find more effective treatment options. Medicinal plants of Malaysia, Middle East, and North Africa (MENA region) following: Algeria, Bahrain, Djibouti, Egypt, Iran, Iraq, Jordan, Kuwait, Lebanon, Libya, Malta, Morocco, Oman, Qatar, Saudi Arabia, Syria, Tunisia, Turkey, United Arab Emirates, Palestine, Yemen, Ethiopia, and Sudan have a significant role in providing new therapeutic remedies. Locals reported that indigenous formulations of some herbs are utilized to treat respiratory disease symptoms relate to TB disease. Deep eutectic solvents (DESs) are considered green solvents and have more significant than traditional solvents in ecosystem. The present study aims to evaluate the potential anti-TB from plants of Malaysia and MENA region extracted by DESs and exploit the traditional medicinal practices of the local people. Aqueous extracts and different types of deep eutectic solvents extracts of 5 kind plant species (Moringa oleifera Lam, Premna odorata Blanco, Ocimum basilicum L, Rosmarinus officinalis L and Eucalyptus camaldulensis) were screened using different assay against multidrug-resistant (MDR) Mycobacterium tuberculosis strain isolated from a clinical sample. The effect of the most active extracts on mycobacterial cells at the cellular level was investigated upon treatment with the crude extracts via time-kill analysis, leakage of compound was absorbing at 280nm. The in vitro cytotoxicity of most active extracts was evaluated against Rat Embryonic Fibroblasts (REF) cell line using MTT assay. Phytochemical profiling of the extracts was investigated by using liquid chromatography-mass spectrometry (LC-MS) in order to identify the chemical constituents present in the most active extracts. The findings revealed that DES2: Tailor extract of R. officinalis displayed the largest zone of inhibition (DIZ= 17.33 ± 1.15 mm) and exhibited the best antimycobacterial activity with minimum inhibitory concentration (3.12 mg/mL). Followed by DES3: CHGl of R. officinalis showed the good minimum inhibitory concentration (MIC= 6.25 mg/mL). DES2: Tailor extract of R. officinalis showed the lowest minimum bactericidal concentration (MBC= 12.5 mg/mL). At 3-fold of MIC, DES2: Tailor extract of R. officinalis killed the entire bacterial cell within 48 h of exposure by causing the cell lysis and releases of cell constituents when measured cell wall integrity. Cytotoxicity assay revealed that the most active DES extract was non-toxic to the REF cell line. The LC-MS analysis revealed the presence of phytoconstituents like: camphor, α-pinene, quercetin, kaempferol and oleic acid that potentially contribute to the antimycobacterial activity. In conclusion, the results demonstrated that the extracts obtained by DESs have incredible potency in TB treatment and are a greener procedure than conventional extraction solvents. Further studies on the use of DESs of plant extract could lead to the development of new anti-TB drugs

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