Antioxidant and Antibacterial Activity of Different Solvent Extracts of Leaves and Stem of Alyxia reinwardtii Blume


  • Vanishri Kaly Sitthan Department of Biology, Universiti Putra Malaysia, 43400, Serdang, Malaysia
  • Muhamad Salihu Abdallah Department of Biology, Universiti Putra Malaysia, 43400, Serdang, Malaysia; Department of Microbiology, Yobe State University, KM 7, Sir Kashim Ibrahim Way, P.M.B 1144, Damaturu, Yobe State, Nigeria
  • Meenakshii Nallappan Department of Biology, Universiti Putra Malaysia, 43400, Serdang, Malaysia
  • Sang-Ho Choi Korea Research Institute of Bioscience and Biotechnology (KRIBB), 125 Gwahak-ro, Yuseong-gu, Daejeon 306-809, South Korea
  • Jin-Hyub Paik Korea Research Institute of Bioscience and Biotechnology (KRIBB), 125 Gwahak-ro, Yuseong-gu, Daejeon 306-809, South Korea
  • Rusea Go Department of Biology, Universiti Putra Malaysia, 43400, Serdang, Malaysia


Apocynaceae, DPPH, extraction, flavonoid, FRAP, medicinal plant, phenolic compound, TPC


Alyxia reinwardtii Blume (Pulasari) Apocynaceae is being widely used as a traditional medicine in treating various illnesses due to its phenolic, coumarins, lignans, iridoid glycosides alkaloids and flavonoids content. These compounds in A. reinwardtii were reported to be useful as medicinal and agricultural potentials. Hence, A. reinwardtii meets the rationale for antioxidant and antibacterial studies to replace synthetic substances using different solvents on the leaves and stem of A. reinwardtii using cold maceration. The Folin-Ciocalteu reagent assay was used to estimate the phenolic content of extracts. The total flavonoid content was determined using aluminum chloride. The antioxidant capacity of the samples was evaluated by 2,2-diphenyl-1-picrylhydrazyl (DPPH) free radical and Ferric Reducing Antioxidant Power (FRAP) assay. Disc Diffusion Assay (DDA), Minimum Inhibitory Concentration (MIC), and Minimum Bactericidal Concentration (MBC) were conducted to evaluate the antibacterial activity of Escherichia coli and Bacillus cereus growth. Methanol was determined as the most effective solvent due to the highest crude yield obtained, (19.47 ± 2.80 %). Acetone showed the highest TPC content (170.44 ± 10.99 mg GAE/g) while hexane showed the highest TFC content (2957 ± 91.24 mg QE/g). Moreover, for DPPH, methanol inhibited the highest antioxidant (75.81 ± 12.62 %) and hexane for FRAP assay (2278.89 ± 69.47 mg AAE/g). Hexane extract is potentially effective with variable efficiency with DDA (11.17 ± 4.48 mm) while methanol extracts with MIC (0.63 μg/mL) and MBC (2.50 μg/mL) against E. coli. The results indicated that methanolic and hexane extract using cold maceration showed optimum antioxidant and antibacterial activity. The present study advocates that A. reinwardtii was a source of substantial antioxidant and antibacterial agents for potential pharmaceutical applications.


Download data is not yet available.


Metrics Loading ...


Abu Bakar, M.F., Abdul Karim, F. & Perisamy, E. 2015. Comparison of phytochemicals and antioxidant properties of different fruit parts of selected Artocarpus species from Sabah, Malaysia. Sains Malaysiana, 44(3): 355-363. DOI:

Adam, O.A.O., Abadi, R.S.M. & Ayoub, S.M.H. 2019. The effect of extraction method and solvents on yield and antioxidant activity of certain Sudanese medicinal plant extracts. The Journal of Phytopharmacology, 8(5): 248-252. DOI:

Alibi, S., Crespo, D. & Navas, J. 2021. Plant-derivatives small molecules with antibacterial activity. Antibiotics, 10: 231. DOI:

Anokwuru, C.P., Anyasor, G.N., Ajibaye, O. & Okebugwu, P. 2011. Effect of extraction solvents on phenolic, flavonoid and antioxidant activities of three Nigerian medicinal plants. Nature and Science of Sleep, 9(7): 53-61.

Anwar, F., Kalsoom, U., Sultana, B., Mushtaq, M., Mehmood, T. & Arshad, H.A. 2013. Effect of drying method and extraction solvent on the total phenolics and antioxidant activity of cauliflower (Brassica oleracea L.) extracts. International Food Research Journal, 20(2): 653-659.

Anwar, M.M., Parveen, A., Hossain, M.M., Mahamud, N.U. & Roy, R.K. 2015. Efficacy of fungicides in controlling late blight of potato. Progressive Agriculture, 26: 103-108. DOI:

Aryal, S., Baniya, M.K., Danekhu, K., Kunwar, P., Gurung, R. & Koirala, N. 2019. Total phenolic content, flavonoid content and antioxidant potential of wild vegetables from Western Nepal. Plants, 8(4): 96. DOI:

Asghar, N., Naqvi, S.A.R., Hussain, Z., Rasool, N., Khan, Z.A., Shahzad, S.A., Sherazi, T.A., Janjua, M. R. S.A., Nagra, S.A., Zia-UI-Haq, M. & Jaafar, H.Z. 2016. Compositional difference in antioxidant and antibacterial activity of all parts of the Carica papaya using different solvents. Chemistry Central Journal, 10: 5. DOI:

Balouiri, M., Sadiki, M. & Ibnsouda, S.K. 2016. Methods for in vitro evaluating antimicrobial activity: A review. Journal of Pharmaceutical Analysis, 6: 71-79. DOI:

Banjarnahor, S.D. & Artanti, N. 2014. Antioxidant properties of flavonoids. Medical Journal of Indonesia, 23(4): 239-44. DOI:

Boeing, J.S., Barizão, É.O., e Silva, B.C., Montanher, P.F., de Cinque Almeida, V. & Visentainer, J.V. 2014. Evaluation of solvent effect on the extraction of phenolic compounds and antioxidant capacities from the berries: Application of principal component analysis. Chemistry Central Journal, 8: 48. DOI:

Breijyeh, Z., Jubeh, B. & Karaman, R. 2020. Resistance of gram-negative bacteria to current antibacterial agents and approaches to resolve it. Molecules, 25(6): 1340. DOI:

Chaves, J.O., de Souza, M.C., da Silva, L.C., Lachos-Perez, D., Torres-Mayanga, P.C., Machado, A., Forster-Carneiro, T., Vázquez-Espinosa, M., González-de-Peredo, A.V., Barbero, G.F. & Rostagno, M.A. 2020. Extraction of flavonoids from natural sources using modern techniques. Frontiers in Chemistry, 8: 507887. DOI:

Choudhary, R.K. & Swarnkar, P.L. 2011. Antioxidant activity of phenolic and flavonoid compounds in some medicinal plants of India. Natural Product Research, 25(11): 1101-1109. DOI:

Cosme, P., Rodríguez, A. B., Espino, J. & Garrido, M. 2020. Plant phenolics: Bioavailability as a key determinant of their potential health-promoting applications. Antioxidants, 9(12): 1263. DOI:

Dirar, A.I., Alsaadi, D.H.M., Wada, M., Mohamed, M.A., Watanabe, T. & Devkota, H.P. 2018. Effects of extraction solvents on total phenolic and flavonoid contents and biological activities of extracts from Sudanese medicinal plants. South African Journal of Botany, 120: 261-267. DOI:

Do, Q.D., Angkawijaya, A.E., Tran-Nguyen, P.L., Huynh, L.H., Soetaredjo, F.E., Ismadji, S. & Yi-Hsu, J. 2014. Effect of extraction solvent on total phenol content, total flavonoid content and antioxidant activity of Limnophila aromatica. Journal Food and Drug Analysis, 22(3): 296-302. DOI:

Dyrda, G., Boniewska-Bernacka, E., Man, D., Barchiewicz, K. & Slota, R. 2019. The effect of organic solvents on selected microorganisms and model liposome membrane. Molecular Biology Reports, 46: 3225-3232. DOI:

Feduraev, P., Chupakhina, G., Maslennikov, P., Tacenko, N. & Skrypnik, L. 2019. Variation in phenolic compounds content and antioxidant activity of different plant organs from Rumex crispus L. and Rumex obtusifolius L. at different growth stages. Antioxidants, 8(7): 237. DOI:

Ghasemzadeh, A., Jaafar, H.Z., Rahmat, A. & Ashkani, S. 2015. Secondary metabolites constituents and antioxidant, anticancer and antibacterial activities of Etlingera elatior (Jack) RM Sm grown in different locations of Malaysia. BMC Complementary and Alternative Medicine, 15(1): 335. DOI:

Hadadi, Z., Nematzadeh, G.A., & Ghahari, S. 2020. A study on the antioxidant and antimicrobial activities in the chloroformic and methanolic extracts of 6 important medicinal plants collected from North of Iran. BMC Chemistry, 14: 33. DOI:

Herrera-Pool, E., Ramos-Díaz, A.L., Lizardi-Jiménez, M.A., Pech-Cohuo, S., Ayora-Talavera, T., Cuevas-Bernardino, J.C., García-Cruz, U. & Pacheco, N. 2021. Effect of solvent polarity on the Ultrasound assisted extraction and antioxidant activity of phenolic compounds from habanero pepper leaves (Capsicum chinense) and its identification by UPLC-PDA-ESI-MS/MS. Ultrasonics Sonochemistry, 76. DOI:

Hien, V.T., Lin, C., Thanh, V.C., Oanh, N.T.K., Thanh, B.X., Weng, C.E., Yuan, C.S. & Rene, E.R. 2019. An overview of the development of vertical sampling technologies for ambient volatile organic compounds (VOCs). Journal of Environmental Management, 247: 401-412. DOI:

Hossain, M.A., Arafat, M.Y., Alam, M. & Hossain, M.M. 2021. Effect of solvent types on antioxidant activity and total flavonoids of some Bangladeshi legumes. Food Research, 5(4): 329-335. DOI:

Iloki-Assanga, S.B., Lewis-Luján, L.M., Lara-Espinoza, C.L., Gil-Salido, A.A., Fernandez- Angulo, D., Rubio-Pino, J.L. & Haines, D.D. 2015. Solvent effects on phytochemical constituent profiles and antioxidant activities, using four different extraction formulations for analysis of Bucida buceras L. and Phoradendron californicum Complementary and Alternative Medicine. BMC Research Notes, 8: 396. DOI:

Inglin, R.C., Stevens, M.J.A., Meile, L., Lacroix, C. & Meile, L. 2015. High-throughput screening assays for antibacterial and antifungal activities of Lactobacillus species. Journal of Microbiological Methods, 114: 26-29. DOI:

Justine, V.T., Mustafa, M., Kankara, S.S. & Go, R. 2019. Effect of drying methods and extraction solvents on phenolic antioxidants and antioxidant activity of Scurrula ferruginae (Jack) Danser (Loranthaceae) leaf extracts. Sains Malaysiana, 48(7): 1383-1393. DOI:

Kumar, S. & Pandey, A.K. 2013. Chemistry and biological activities of flavonoids: An overview. The Scientific World Journal, 2013: 162750. DOI:

Lin, D., Xiao, M., Zhao, J., Li, Z., Xing, B., Li, X., Kong, M., Li, L., Zhang, Q., Liu, Y., Chen, H., Qin, W., Wu, H. & Chen, S. 2016. An overview of plant phenolic compounds and their importance in human nutrition and management of type 2 diabetes. Molecules, 21(1): 3. DOI:

Mahasneh, A.M. & Al-Hussaini, R. 2011. Antibacterial and antifungal activity of ethanol extract of different parts of medicinal plants in Jordan. Jordan Journal of Pharmaceutical Sciences, 108(397): 1-26.

Marasini, B.P., Baral, P., Aryal, P., Ghimire, K.R., Neupane, S., Dahal, N., Singh, A., Ghimire, L. & Shrestha, K. 2015. Evaluation of antibacterial activity of some traditionally used medicinal plants against human pathogenic bacteria. BioMed Research International, 2015: 265425. DOI:

Mathesius, U. 2018. Flavonoid functions in plants and their interactions with other organisms. Plants, 7(2): 7-9. DOI:

Mohamed, A.A., Ali, S.I. & El-Baz, F.K. 2013. Antioxidant and antibacterial activities of crude extracts and essential oils of Syzygium cumini leaves. PLoS ONE, 8(4): e60269. DOI:

Monteiro, M., Santos, R.A., Iglesius, P., Couto, A., Serre, C. R., Gouvinhas, I., Barros, A., Olivia-Teles, A., Enes, P. & Díaz-Rosales, P. 2020. Effect of extraction method and solvent system on the phenolic content and antioxidant activity of selected macro- and microalgae extracts. Journal of Applied Phycology, 32: 349-362. DOI:

Muskhazli, M., Dirnahayu, M., Nor Azwady, A.A., Nurhafiza, Y., Nor, D.E. & Che Ku Nurshaira, C.K.N. 2009. Antibacterial activity of methanolic crude extracts from selected plant against Bacillus cereus. Pertanika Journal of Tropical Agricultural Science, 32(2): 175-183.

Nadia, Z. & Rachid, M. 2013. Antioxidant and antibacterial activities of thymus vulgaris. Medicinal and Aromatic Plant Research Journal, 1: 5-11.

Nagwa, M.A., Sanaa, M.S., Sahar, I.N. & Yasmeen, A.A. 2019. Evaluation of antimicrobial activity of some plant extracts against antibiotic susceptible and resistant bacterial strains causing wound infection. Bulletin of the National Research Centre, 43: 144. DOI:

Ngo, T.V., Scarlett, C.J., Bowyer, M.C., Ngo, P.D. & Vuong, Q.V. 2017. Impact of different extraction solvents on bioactive compounds and antioxidant capacity from the root of Salacia chinensis L. Journal of Food Quality, 2017: 9305047. DOI:

Njoya, E.M. 2021. Chapter 31- medicinal plants, antioxidant potential, and cancer. In: Cancer. V.R. Preedy and V.B. Patel (Eds.). Academic Press. pp. 349-357. DOI:

Nugroho, A.E., Wijayanti, A., Mutmainah, M., Susilowati, R. & Rahmawati, N. 2016. Gastroprotective effect of combination of hot water extracts of Licorice (Glycyrrhiza glabra), Pulasari (Alyxia reinwardtii) and Sembung leaf (Blumea balsamifera) against aspirin-induced gastric ulcer model rats. Journal of Evidence-Based Complementary & Alternative Medicine, 21(4): 77-84. DOI:

Obeidat, M., Shatnawi, M., Al-alawi, M., Al-Zu'bi, E., Al-Dmoor, H., Al-Qudah, M., El-Qudah, J. & Otri, I. 2012. Antimicrobial activity of crude extracts of some plant leaves. Research Journal of Microbiology, 7(1): 59-67. DOI:

Ortega-Ramirez, L.A., Rodriguez-Garcia, I., Leyva, J.M., Cruz-Valenzuela, M.R., Silva Espinoza, B.A., Gonzalez-Aguilar, G.A., Siddiqui, M.W. & Ayala-Zavala, J.F. 2014. Potential of medicinal plants as antimicrobial and antioxidant agents in food industry: A hypothesis. Journal of Food Science, 79(2). DOI:

Panche, A.N., Diwan, A.D. & Chandra, S.R. 2016. Flavonoids: An overview. Journal of Nutritional Science, 5: e47. DOI:

Peng-ngummuang, K., Palanuvej, C. & Ruangrungsi, N. 2015. Pharmacognostic specification and coumarin content of Alyxia reinwardtii inner bark. Engineering Journal, 19(3): 15. DOI:

Pham-Huy, L. A., He, H. & Pham-Huy, C. 2008. Free radicals, antioxidant in disease and health. International Journal of Biomedical Science, 4(2): 89-96. DOI:

Procházková, D., Boušová, I. & Wilhelmová, N. 2011. Antioxidant and prooxidant properties of flavonoids. Fitoterapia, 82(4): 513-523. DOI:

Rafat, A., Philip, K. & Muniandy, S. 2010. Antioxidant potential and content of phenolic compounds in ethanolic extracts of selected parts of Andrographis paniculata. Journal of Medicinal Plants Research, 4(3): 197-202.

Rattanapan, J., Sichaem, J. & Tip-yang, S. 2012. Chemical constituents and antioxidant activity from the stems of Alyxia reinwardtii. Records of Natural Products, 6(3): 288-291.

Routray, W. & Orsat, V. 2012. Microwave-assisted extraction of flavonoids: A review. Food and Bioprocess Technology, 5(2), 409-424. DOI:

Sahoo, C.R., Sahoo, J., Mahapatra, M., Lenka, D., Sahu, P.K., Dehury, B., Padhy, R.N. & Paidesetty, S.K. 2021. Coumarin derivatives as promising antibacterial agent(s). Arabian Journal of Chemistry, 14(2): 102922. DOI:

Saiah, H., Allem, R. & El Kebir, F.Z. 2016. Antioxidant and antibacterial activities of six Algerian medicinal plants. International Journal of Pharmacy and Pharmaceutical Sciences, 8(1): 367-374.

Samanta, A., Das, G. & Das, S. 2011. Roles of flavonoids in plants. Carbon, 6(1): 12-35.

Sankeshwari, R.M., Ankola, A.V., Bhat, K. & Hullatti, K. 2018. Soxhlet versus cold maceration: Which method gives better antimicrobial activity to Liquorice extract against Streptococcus mutans?. Journal of the Scientific Society, 45(2): 67-71. DOI:

Santo-Sánchez, N.F., Salas-Coronado, R., Hernández-Carlos, B. & Villanueva-Cañongo, C. 2019. Shikimic acid pathway in biosynthesis of phenolic compounds. In: Plant Physiological Aspects of Phenolic Compounds. M. Soto-Hernández, R. García-Mateos and M. Palma-Tenango (Eds.). IntechOpen.

Sepahpour, S., Selamat, J., Manap, M.Y.A., Khatib, A. & Razis, A.F.A. 2018. Comparative analysis of chemical composition, antioxidant activity and quantitative characterizationof some phenolic compounds in selected herbs and spices in different solvent extraction systems. Molecules, 23(2): 402. DOI:

Shahidi, F. & Zhong, Y. 2015. Measurements of antioxidant activity. Journal of Functional Foods, 18: 757-781. DOI:

Stankovic, M.S., Niciforovic, N., Topuzovic, M. & Solujic, S. 2011. Total phenolic content, flavonoid concentrations, and antioxidant activity, of the whole plant and plant parts extracts from Teucrium montanum L. var. montanum, F. supinum (L.) Reichenb. Biotechnology and Biotechnological Equipment, 25(1): 2222-2227. DOI:

Sugijanto, N.E., Diesel, A., Ebel, R., Indrayanto, G. & Zaini, N.C. 2009. Chemical constituents of the endophytic fungus Lecythophora sp. isolated from Alyxia reinwardtii. Natural Product Communications, 4(11): 1485-1488. DOI:

Sumayya, T.K. & Gopinathan, M. 2022. Correlation of antioxidant, antibacterial and phytochemicals of leaves and roots extracts of Clerodendrum infortunatum. International Journal of Pharmaceutical Sciences and Research, 13(5): 2144-2153.

Tan, M.C., Tan, C.P. & Ho, C.W. 2013. Effects of extraction solvent system, time and temperature on total phenolic content of henna (Lawsonia inermis) stems. International Food Research Journal, 20(6): 3117.

Topçu, G., Che, C., CordellĦ, G.A. & Ruangrungsi§, N. 1990. Iridolactones from Alyxia reinwardtii. Phytochemistry, 29(10): 3197-3199. DOI:

Wang, S.Y., Chen, H., Camp, Mary, J.C. & Mart, K.E. 2012. Flavonoid constituents and their contribution to antioxidant activity in cultivars and hybrids of Rabbit Eye Blueberry (Vaccinium ashei Reade). Food Chemistry, 132(2): 855-864. DOI:

Wijekoon, M.M.J.O., Bhat, R. & Karim, A.A. 2011. Effect of extraction solvents on the phenolic compounds and antioxidant activities of bunga kantan (Etlingera elatior Jack.) inflorescence. Journal of Food Composition and Analysis, 24: 615-619. DOI:

Zlotek, U., Mikulska, S., Nagajek, M. & Świeca, M. 2016. The effect of different solvents and number of extraction steps on the polyphenol content and antioxidant capacity of basil leaves (Ocimum basilicum L.) extracts. Saudi Journal of Biological Sciences, 23: 628-633. DOI:



How to Cite

Sitthan, V. K., Abdallah, M. S. ., Nallappan, M. ., Choi, S.-H. ., Paik, J.-H. ., & Go, R. (2023). Antioxidant and Antibacterial Activity of Different Solvent Extracts of Leaves and Stem of Alyxia reinwardtii Blume. Malaysian Applied Biology, 52(6), 67–80.



Research Articles