Comparative Analysis of Phytochemical Composition in Ethanolic Extracts of Acalypha Indica Between Ultrasound-Assisted and Maceration Extraction Methods
Keywords:
Acalypha indica, Ultrasonic-Assisted Extraction, Maceration Extraction, Polyphenol, AntioxidantAbstract
Acalypha indica (AI) is one of the edible traditional medicinal herbs used worldwide. Previously, conventional maceration extraction was often used to extract phytochemicals from AI plant material with limitations of prolonged extraction time and high solvent consumption. This study investigates the extraction methods, namely maceration extraction (ME) and ultrasound-assisted extraction (UAE), which were utilised to extract and compare the phytochemical content and antioxidant activity of the AI sample derived from the whole plant. The extraction involved the use of water solvent and ethanolic solvent (20%, 40%, 60%, and 80%). Each extract was subjected to chemical-assay guidance by testing for Total Phenolic Content (TPC), Total Flavonoid Content (TFC), and 2,2-diphenyl-1-picrylhydrazyl (DPPH) radical scavenging assay. The study reveals UAE extract gained a low EC50 (84.5 μg/ml) at 40% ethanolic solvent, the highest TPC (555.952 ± 41.938 mg GAE/g) using water solvent, and 60% obtained the highest TFC (887.9048 ± 46.085 mg RE/g). As for maceration, 60% obtained a low value of EC50 (115.1 μg/ml) and the highest TPC (567.061 ± 16.432 mg GAE/g). Meanwhile, 80% ethanol solvent obtained the highest TFC (577.619 ± 43.008 mg RE/g). Ultimately, maceration achieved higher polyphenolic content while having the better antioxidant outcome. However, the UAE extracted polyphenolic content in a shorter period of time and with lower solvent consumption.
References
Abd Rashid, S. N. A., Hasham, R., Abd. Rashid, Z. I., Cheng, K. K., Aziz, A. A., Shafin, N., & Kaprawi, A. A. (2022). Formulation and characterization of the physicochemical, antioxidant activity and sensory attributes of curcuma-based herbal drink. Materials Today: Proceedings, 57, 1061–1066.
Adeeyo, A. O., Oyetade, J. A., Alabi, M. A., Adeeyo, R. O., Samie, A., & Makungo, R. (2023). Tuning water chemistry for the recovery of greener products: pragmatic and sustainable approaches. RSC Advances, 13(10), 6808–6826.
Akshayaa, L., Malaippan, D., & Rajeshkumar, S. (2021). Evaluation of antioxidant and anti-inflammatory activity of Acalypha indica aqueous ethanolic extract. Nveo – Natural Volatiles & Essential Oils Journal | NVEO, 5965– 5977.
Borges, A., José, H., Homem, V., & Simões, M. (2020). Comparison of techniques and solvents on the antimicrobial and antioxidant potential of extracts from Acacia dealbata and Olea europaea. Antibiotics, 9(2), 48.
Brglez Mojzer, E., Knez Hrnčič, M., Škerget, M., Knez, Ž., & Bren, U. (2016). Polyphenols: Extraction methods, antioxidative action, bioavailability and anticarcinogenic effects. Molecules, 21(7), 901.
Carmen, A. L., Cristina, G., Elena, P. A., & Eufrosina, A. (2022). Extraction of active compounds from Armoracia rusticana using maceration and ultrasound assisted extraction. 2022 E-Health and Bioengineering Conference (EHB).
Debiasi, B. W., Rodrigues, P. G. R. S., Torres, M. P. R., Bonacorsi, C., Andrighetti, C. R., Ribeiro, E. B., & Valladão, D. M. S. (2021). Comparison between maceration and ultrasound-assisted extraction of white bracts with flowers of Bougainvillea spectabilis Willd. Scientific Electronic Archives, 14(2), 47.
Gantala, S. R., Kalukuri, S., Dommat, W. G., Volukula, V., Gangi, S., Saval, V., & Koppula, P. (2023). Therapeutic and biological aspects of leaf extracts from indian copper leaf plant (Acalypha Indica). International Journal of Plant and Environment, 9(02), 150–156.
Gulcin, İ. (2020). Antioxidants and antioxidant methods: an updated overview. Archives of Toxicology, 94(3), 651–715.
Gülçin, İ., & Alwasel, S. (2023). DPPH radical scavenging assay. Processes, 11(8), 2248–2248.
Huynh, T. T. H., Wongmaneepratip, W., & Vangnai, K. (2025). Relationship between flavonoid chemical structures and their antioxidant capacity in preventing polycyclic aromatic hydrocarbons formation in heated meat model system. Foods, 13(7).
Ibrahim, A. M., Hamid, M. A., Althiab, R. A., Amir, & Zulkifli, R. M. (2021). In vitro fibroblasts viability and migration stimulation of Acalypha indica: an insight on wound healing activity. Future Journal of Pharmaceutical Sciences, 7(1).
Idris, M. K. H., Hasham, R., & Ismail, H. F. (2022). Bioassay-guided extraction of andrographis paniculata for intervention of in-vitro prostate cancer progression in metabolic syndrome environment. Daru: Journal of Faculty of Pharmacy, Tehran University of Medical Sciences, 30(2), 253–272.
Ismail, H. F., Hashim, Z., Soon, W. T., Rahman, N. S. A., Zainudin, A. N., & Majid, F. A. A. (2017). Comparative study of herbal plants on the phenolic and flavonoid content, antioxidant activities and toxicity on cells and zebrafish embryo. Journal of Traditional and Complementary Medicine, 7(4), 452–465.
Jacotet-Navarro, M., Laguerre, M., Fabiano-Tixier, A.-S., Tenon, M., Feuillère, N., Bily, A., & Chemat, F. (2018). What is the best ethanol-water ratio for the extraction of antioxidants from rosemary? Impact of the solvent on yield, composition, and activity of the extracts. Electrophoresis, 39(15), 1946–1956. https://doi.org/10.1002/elps.201700397
Kedare, S. B., & Singh, R. P. (2011). Genesis and development of DPPH method of antioxidant assay. Journal of Food Science and Technology, 48(4), 412–422.
Koksal, E., Bursal, E., Dikici, E., Tozoglu, F., & Gulcin, I. (2011). Antioxidant activity of Melissa officinalis leaves. Journal of Medicinal Plants Research, 5(2), 217–222.
Kumar, K., Srivastav, S., & Sharanagat, V. S. (2021). Ultrasound assisted extraction (UAE) of bioactive compounds from fruit and vegetable processing by-products: A review. Ultrasonics Sonochemistry, 70(70), 105325.
Kusrini, D., Fachriyah, E., & Prinanda, G. R. (2019). Isolation of phenolic acid in Acalypha indica l plants and test total phenol also antioxidant test using DPPH method. IOP Conference Series: Materials Science and Engineering, 509, 012033.
Lim, K. J. A., Cabajar, A. A., Lobarbio, C. F. Y., Taboada, E. B., & Lacks, D. J. (2019). Extraction of bioactive compounds from mango (Mangifera indica L. var. Carabao) seed kernel with ethanol–water binary solvent systems. Journal of Food Science and Technology, 56(5), 2536–2544.
Majid, H., & Silva, F. V. M. (2021). Kanuka bush leaves for Alzheimer’s disease: Improved inhibition of β-secretase enzyme, antioxidant capacity and yield of extracts by ultrasound assisted extraction. Food and Bioproducts Processing, 128, 109–120.
Mikucka, W., Zielińska, M., Bułkowska, K., & Witońska, I. (2022). Valorization of distillery stillage by polyphenol recovery using microwave-assisted, ultrasound-assisted and conventional extractions. Journal of Environmental Management, 322, 116150.
Morales-Olán, G., Rojas‐López, M., Díaz-Reyes, J., De Fátima Rosas-Cárdenas, F., & Luna- Suárez, S. (2020). Effect of ethanol and methanol on the total phenolic content and antioxidant capacity of chia seeds (Salvia hispanica L.). Sains Malaysiana, 49(6), 1283–1292.
N’Paki, K. M., Domedjui, K. L., Melila, M., Kanda, M., Koffi Apeti Gbogbo, K. A., Folega, F., Dourma, M., Batawila, K., & Akpagana, K. (2024). Phytochemical characterization, antioxidant and antimicrobial properties of hydroethanolic extract from Acalypha indica L. (Euphorbiaceae) collected in Togo. Asian Journal of Biology, 20(7), 50–63.
Nambiar, M. K. G., & Varghese, V. (2023). In vitro hepatoprotective activity of methanolic leaf extract of Acalypha indica against CCl4 induced hepatotoxicity in goat liver slice culture. Trends in Sciences, 20(4), 4562.
Ngibad, K. (2019). Phytochemical screening of sunflower leaf (Helianthus annuus) and anting-anting (Acalypha indica Linn) plant ethanol extract. Borneo Journal of Pharmacy, 2(1), 24–30.
Nurcholis, W., Alfadzrin, R., Izzati, N., Arianti, R., Vinnai, B. Á., Sabri, F., Kristóf, E., & Artika, I. M. (2022). Effects of methods and durations of extraction on total flavonoid and phenolic contents and antioxidant activity of Java Cardamom (Amomum compactum Soland Ex Maton) fruit. Plants, 11(17), 2221–2221.
Nyero, A., Anywar, G. U., Achaye, I., & Malinga, G. M. (2023). Phytochemical composition and antioxidant activities of some wild edible plants locally consumed by rural communities in northern Uganda. Frontiers in Nutrition, 10.
Ravi, S., Shanmugam, B., Prasad, S. H., Ganjikunta Venkata Subbaiah, & Reddy, K. S. (2015). Assessment of potential antioxidant activity of polyphenolic fraction separated from Acalypha Indica leaves: An in vitro approach. International Journal of Pharma Research and Review, 2015.
Ribeiro, A., Romeiras, M. M., Tavares, J., & Faria, M. T. (2010). Ethnobotanical survey in Canhane village, district of Massingir, Mozambique: Medicinal plants and traditional knowledge. Journal of Ethnobiology and Ethnomedicine, 6(1).
Safe, S., Jayaraman, A., Chapkin, R. s, Howard, M., Mohankumar, K., & Shrestha, R. (2021). Flavonoids: structure-function and mechanisms of action and opportunities for drug development. Toxicology Research, 37(2).
Sahukari, R., Punabaka, J., Bhasha, S., Ganjikunta, V. S., Kondeti Ramudu, S., Kesireddy, S. R., Ye, W., & Korivi, M. (2021). Phytochemical profile, free radical scavenging and anti-inflammatory properties of Acalypha indica root extract: Evidence from In vitro and in vivo studies. Molecules, 26(20), 6251.
Salma Iza, F., Hayati, E. K., Rafi, M., & Sabarudin, A. (2023). Effect of ethanol-water concentration as extraction solvent on antioxidant activity of Acalypha indica. Al Kimiya: Jurnal Ilmu Kimia Dan Terapan, 10(2).
Sarabo, Z., Hanafi, N., Azzuan Rosli, M., Norazlina Abd Rashid, S., Mohd Ropi, N. A., Hasham, R., Sarmidi, M. R., Cheng, K. K., & Othman, N. Z. (2021). Effect of different pre-treatments on the physicochemical properties of Freeze-Dried Ananas comosus L. Materials Today: Proceedings, 42(1), 229–233.
Scaffidi, A., Algar, D., Bohman, B., Ghisalberti, E. L., & Flematti, G. (2016). Identification of the cat attractants isodihydronepetalactone and isoiridomyrmecin from Acalypha indica. Australian Journal of Chemistry, 69(2), 169.
Sharma, D., Sharma, S., Mandal, V., & Dhobi, M. (2023). Unveiling the anti-inflammatory potential of Acalypha indica L. and analyzing its research trend: digging deep to learn deep. Naunyn-Schmiedeberg’s Archives of Pharmacology, 397.
Teffane, M., Boudries, H., Bey, M. B., Kadi, A., & Boukhalfa, F. (2022). Effect of solvent type, extraction temperature, agitation speed and microwave power on phenolic compound extraction and antioxidant activity of apricot kernels (Prunus armeniaca L.). Current Bioactive Compounds, 18(1).
Venkatesan, U., & Muniyan, R. (2025). Therapeutic potential of Acalypha indica L. leaf fractions against foodborne pathogens: an in vitro and in silico study. Scientific Reports.
Venkatesan, T., Choi, Y.-W., & Kim, Y.-K. (2019). Impact of different extraction solvents on phenolic content and antioxidant potential of pinus densiflora 70 bark extract. BioMed Research International, 2019, 1–14.
Wickramaratne, S., de Silva, N., Wickramasinghe, S., Wanigasekara, A., Adhikari, R., & Rajapakasha, E. (2022). Olfactory behaviour reactions to Acalypha indica preparations in domestic cats. Applied Animal Behaviour Science, 257, 105776.
Yıldırım, H. K. (2022). Assessment of propolis treated by different extraction methods. Brazilian Archives of Biology and Technology, 65.
Zahidin, N. S., Saidin, S., Zulkifli, R. M., Muhamad, I. I., Ya’akob, H., & Nur, H. (2017). A review of Acalypha indica L. (Euphorbiaceae) as traditional medicinal plant and its therapeutic potential. Journal of Ethnopharmacology, 207, 146–173.
Zahidin, N. S., Zulkifli, R. M., Muhamad, I. I., Ya’akob, H., Nur, H., Shariff, A. H. M., & Saidin, S. (2018). Preliminary study of potential herbal tea, Acalypha indica and comparison with domestic tea in Malaysia market. Food Science and Technology, 6(1), 41–45.
Ziani, I., Bouakline, H., Yahyaoui, M. I., Belbachir, Y., Fauconnier, M.-L., Asehraou, A., Tahani, A., Talhaoui, A., & El Bachiri, A. (2022). The effect of ethanol/water concentration on phenolic composition, antioxidant, and antimicrobial activities of Rosmarinus tournefortii de Noé hydrodistillation solid residues. Journal of Food Measurement and Characterization, 17(2), 1602–1615.
