2024(2)      April- June, 2024

Characterization and identification of bioactive natural products in the ethanol extracts of Acacia nilotica, Melia azedarach, and Euphorbia hirta from Cholistan desert, Pakistan

Sobia Malik1,2, Nuzhat Sial1, Mirza Imran Shahzad3*, Shazia Anjum4, Arshad Javid5, Gildardo Rivera6

1Department of Zoology, The Islamia University, Bahawalpur, Pakistan

2Department of Zoology, Government Sadiq College Women University, Bahawalpur, Pakistan

3Department of Biochemistry, The Islamia University, Bahawalpur, Pakistan

4Department of Chemistry, The Islamia University, Bahawalpur, Pakistan

5Department of Wildlife and Ecology, University of Veterinary and Animal Sciences, Lahore, Pakistan

6Laboratorio de Biotecnología Farmacéutica, Centro de Biotecnología Genómica, Instituto Politécnico Nacional, México



Cholistan desert plants form Fabaceae, Meliaceae and Euphorbiaceae families have always been recognized as an alternate source of medicine and used in different pharmacological activities due to the presence of bioactive secondary metabolites. This study was aimed to characterize bioactive contents in ethanol extracts of Acacia nilotica (whole branch, bark), Melia azedarach (leaves, bark,) and Euphorbia hirta (whole plant). Characterization and composition of secondary metabolites were determined by both chromatographic and non-chromatographic techniques. TLC profile showed maximum spots in Acacia nilotica and M. azedarach. A. nilotica whole branch yielded nine spots for n-Hex, seven spots for DCM, 4 spots for EtAC while A. nilotica bark yielded 4 spots for n-Hex, nine spots for DCM and 4 spots for EtAC. M. azedarach leaves revealed seven spots for n-Hex, six spots for DCM, seven spots for EtAC, while M. azedarach bark revealed seven spots for n-Hex, five spots for DCM, and also five spots for EtAC. E. hirta yielded six spots for n-Hex, two spots for DCM and for EtAC two spots detected. FT-IR spectra showed the characteristic prominent peaks. The maximum number of functional groups were observed in M. azedarach bark, followed by A. nilotica whole branch/bark and M. azedarach whole branch. The least number of functional groups were observed in E. hirta. HPLC analysis was revealed that 9 compounds were majorly quantified in A. nilotica whole branch bark i.e., Gallic Acid, p-hydroxy benzoic acid, Gentisic Acid, Protocatechuic Acid, Catechin, Syringic Acid, Chlorogenic Acid, Vanillic acid, and Epi-catechin. twenty-three compounds were predominantly quantified in M. azedarach leaves bark i.e., Quercetin, Hydroxy ferulic acid hexoside, Rutin, Vanillic Acid, Ferulic Acid, Ferulic acid hexoside II, Feruloylquinic Acid, Myricetin hexoside, Kaempferol -3- O -rutinoside, Kaempferol -3- O – rhamnoside, Procyanidin dimer B, Toosendanin, Quercetin-7-O- glycoside, Kaempferol, Catechin-7-O- glycoside, Apigenin -7-O- glycoside, Kaempferol -7-O-glycoside, Catechin-5-O- glycoside, Capric acid methyl ester, 8- Hexadecene, Phytadiene, γ-n-Amyl butyrolactone, Apigenin, Luteolin, Kaempferol -3-O-glycoside and from E. hirta seven compounds were quantified i.e., Rutin, Gallic Acid, Tannic Acid, Resorcinol, Ellagic Acid, Benzoic Acid, Quercetin. The LCMS scan of A. nilotica whole branch demonstrated the presence of twelve active compounds showing 1.612 -11.183 retention time, fifteen compounds confirmed in A. nilotica bark with rt 0.700 – 14.202, M. azedarach leaves showed only eleven compounds with rt 1.494 -13.031, M. azedarach bark showed 14 compounds in with rt 1.659 – 13.039 and E. hirta revealed eleven compounds with rt 1.557 – 10.884. The GCMS scan of A. nilotica whole branch ten compounds were detected with rt 23.529 – 35.779, A. nilotica bark 9 compounds identified with rt 6.180 – 36.157. Only one compound from M. azedarach leaves identified with rt 34.116, while 5 compounds found in M. azedarach bark with rt 30.740 – 35.379. E. hirta revealed twenty-eight compounds with rt 6.411 – 36.933. The experimental data of studies suggest that the presence of active compound introduce the therapeutic use against infectious diseases and also based on assumption that Cholistan desert medicinal plants are rich source(s) that confers various biological activities.


Keywords: Cholistan desert, Medicinal plants, Ethanol extraction, Liquid-liquid fractionation, TLC, Phytochemical analysis

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