APPLICATION OF MIXTURE DESIGN TO ENHANCE FLAVONOID CONTENT AND IDENTIFY SECONDARY METABOLITES IN PORTULACA OLERACEA L.
DOI:
https://doi.org/10.55766/sujst6704Keywords:
Antioxidant, LC-HR/MS, Simplex Centroid Mixture Design, Total Flavonoid ContentAbstract
Flavonoids are secondary plant metabolites with numerous pharmacological benefits. However, selecting an appropriate extraction solvent is crucial for effectively extracting flavonoids from plants. Portulaca oleracea L., a wild plant rich in flavonoids, offers significant potential for development through solvent extraction techniques. Therefore, this study aims to enhance the total flavonoid content (TFC) of P. oleracea extracts using a simplex centroid mixture design (SCMD) and to identify the secondary metabolites present. SCMD was formulated using Design Expert v.13 software. Various extraction solvents, including water, acetone, methanol, and ethanol, served as independent variables, while the response variable was the TFC, measured through the AlCl3 colorimetric method. LC-HR/MS analysis was performed to identify the secondary metabolites of the optimal extract. The results showed that an acetone–ethanol mixture produced the highest TFC (2.07 mg QE/g DW), but not significantly different from 100% methanol (1.95 mg QE/g DW) and acetone-methanol (1.88 mg QE/g DW) combinations. A linear model (p < 0.05) was generated from these response formulations. The optimal extraction solvent formula identified was a 50:50 acetone-methanol mixture, with a desirability value of 0.81. Model validation confirmed the reliability of this formula (RSE < 10%). Additionally, the P. oleracea extract showed significant antioxidant capacity, with reducing power against ABTS radicals (21.16 μmol TE/g DW), Fe³⁺ (12.33 μmol TE/g DW), and Cu²⁺ (38.57 μmol TE/g DW). Secondary metabolite analysis also showed the presence of 9 phenols, 8 flavonoids, 5 alkaloids, 5 indoles, and 3 quinolines. These results provide a valuable reference for further studies on the extraction of flavonoid from P. oleracea.
References
Ahmadi, S. M., Farhoosh, R., Sharif, A., & Rezaie, M. (2020). Structure-antioxidant activity relationships of luteolin and catechin. Journal of Food Science, 85(2), 298-305. https://doi.org/10.1111/1750-3841.14994
Aisyah, S. I., Oktavia, A. W. P., Ayuningtyas, A. A., Putra, R. P., Prassiska, S., Jamilah, S., & Nurcholis, W. (2023). Short communication: Differences in phytochemical compounds and antioxidant activity of Portulaca oleracea and Portulaca grandiflora. Biodiversitas, 24(3), 1385-1390. https://doi.org/10.13057/biodiv/d240307
Cavalcante, G. M., Cabral, A. E. da S., & Silva, C. C. (2018). Leishmanicidal activity of natural and synthetic flavonoids: A minireview. Mintage Journal of Pharmaceutical and Medical Sciences, 7, 25-34. https://doi.org/10.37885/220207877
Chen, W. C., Wang, S. W., Li, C. W., Lin, H. R., Yang, C. S., Chu, Y. C., Lee, T. H., & Chen, J. J. (2022). Comparison of various solvent extracts and major bioactive components from Portulaca oleracea for antioxidant, anti-tyrosinase, and anti-α-glucosidase activities. Antioxidants, 11(2), 398. https://doi.org/10.3390/antiox11020398
Dias, M. C., Pinto, D. C. G. A., & Silva, A. M. S. (2021). Plant flavonoids: Chemical characteristics and biological activity. Molecules, 26(17), 5377. https://doi.org/10.3390/molecules26175377
Hwangdee, P., Charee, S., Kheowkrai, W., Junsiri, C., & Laloon, K. (2022). Application of the simplex-centroid mixture design to biomass charcoal powder formulation ratio for biomass charcoal briquettes. Sustainability, 14(7), 3940. https://doi.org/10.3390/su14073940
Idrissi, A., Polok, K., Barj, M., Marekha, B., Kiselev, M., & Jedlovszky, P. (2013). Free energy of mixing of acetone and methanol: A computer simulation investigation. The Journal of Physical Chemistry B, 117(50), 16157-16164. https://doi.org/10.1021/jp405090j
Jdaini, K., Alla, F., Mansouri, F., Parmar, A., & Elhoumaizi, M. A. (2023). Optimizing the extraction of phenolic antioxidants from date palm fruit by simplex-centroid solvent mixture design. Heliyon, 9(1), e12738. https://doi.org/10.1016/j.heliyon.2022.e12738
Kashyap, P., Kumar, S., Riar, C. S., Jindal, N., Baniwal, P., Guiné, R. P. F., Correia, P. M. R., Mehra, R., & Kumar, H. (2022). Recent advances in drumstick (Moringa oleifera) leaves bioactive compounds: Composition, health benefits, bioaccessibility, and dietary applications. Antioxidants, 11(2), 402. https://doi.org/10.3390/antiox11020402
Liga, S., Paul, C., & Péter, F. (2023). Flavonoids: Overview of biosynthesis, biological activity, and current extraction techniques. Plants, 12(14), 2732. https://doi.org/10.3390/plants12142732
Makkiyah, F. A., Rahmi, E. P., Susantiningsih, T., Marliani, N., Arista, R. A., & Nurcholis, W. (2023). Optimization of Graptophyllum pictum leaves extraction using a simplex centroid design focused on extracting flavonoids with antioxidant activity. Journal of Applied Pharmaceutical Science, 13(5), 214-221. https://doi.org/10.7324/japs.2023.117043
Maulana, F., Muhammad, A. A., Umar, A., Mahendra, F. R., Musthofa, M., & Nurcholis, W. (2022). Profiling metabolites through chemometric analysis in Orthosiphon aristatus extracts as α-glucosidase inhibitory activity and in silico molecular docking. Indonesian Journal of Chemistry, 22(2), 501-514. https://doi.org/10.22146/ijc.71334
Ngo, V., & Duennwald, M. L. (2022). Nrf2 and oxidative stress: A general overview of mechanisms and implications in human disease. Antioxidants, 11(12), 2345. https://doi.org/10.3390/antiox11122345
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. https://doi.org/10.3390/plants11172221
Panche, A. N., Diwan, A. D., & Chandra, S. R. (2016). Flavonoids: An overview. Journal of Nutritional Science, 5, e47. https://doi.org/10.1017/jns.2016.41
Pandey, K. B., & Rizvi, S. I. (2009). Plant polyphenols as dietary antioxidants in human health and disease. Oxidative Medicine and Cellular Longevity, 2(5), 270-278. https://doi.org/10.4161/oxim.2.5.9498
Patel, M. B., Shaikh, F., Patel, V., & Surti, N. I. (2017). Application of simplex centroid design in formulation and optimization of floating matrix tablets of metformin. Journal of Applied Pharmaceutical Science, 7(4), 23-30. https://doi.org/10.7324/japs.2017.70403
Ponphaiboon, J., Krongrawa, W., Aung, W. W., Chinatangkul, N., Limmatvapirat, S., & Limmatvapirat, C. (2023). Advances in natural product extraction techniques, electrospun fiber fabrication, and the integration of experimental design: A comprehensive review. Molecules, 28(13), 5163. https://doi.org/10.3390/molecules28135163
Putra, R. P., Aisyah, S. I., & Nurcholis, W. (2023). Benefits of total phenolic and flavonoid content of Portulaca oleracea as antioxidant and antidiabetic: A review. Tropical Journal of Natural Product Research, 7(2), 2293-2304. https://doi.org/10.26538/tjnpr/v7i2.1
Putra, R. P., Aisyah, S. I., Kurniatin, P. A., & Nurcholis, W. (2024). Optimization of solvent sonication–maceration for enhancing total phenolic content and antioxidant activity of Portulaca oleracea L. extract using the simplex centroid design method. Tropical Journal of Natural Product Research, 8(3), 6552-6557. https://doi.org/10.26538/tjnpr/v8i3.12
Rohman, S., & Aini, A. Q. (2023). Total phenolics and flavonoid content of maceration-based purslane (Portulaca oleracea) extract. Nusantara Science and Technology Proceedings, 2023(33), 421-424. https://doi.org/10.11594/nstp.2023.3367
Sajid, M., Channakesavula, C. N., Stone, S. R., & Kaur, P. (2021). Synthetic biology towards improved flavonoid pharmacokinetics. Biomolecules, 11(5), 754. https://doi.org/10.3390/biom11050754
Sicari, V., Loizzo, M. R., Tundis, R., Mincione, A., & Pellicanò, T. M. (2018). Portulaca oleracea L. extracts display antioxidant and hypoglycaemic effects. Journal of Applied Botany and Food Quality, 91, 39-46. https://doi.org/10.5073/JABFQ.2018.091.006
Suhaimi, S. H., Hasham, R., Idris, M. K. H., Ismail, H. F., Ariffin, N. H. M., & Majid, F. A. A. (2019). Optimization of ultrasound-assisted extraction conditions followed by solid phase extraction fractionation from Orthosiphon stamineus Benth. leaves for antiproliferative effect on prostate cancer cells. Molecules, 24(22), 4183. https://doi.org/10.3390/molecules24224183
Thao, T. T. P., Tu, P. T. C., & Men, T. T. (2023). A comparative study on polyphenol and flavonoid content, antioxidant and anti-inflammatory capacity of different solvent extracts from Portulaca oleracea in carrageenan-induced paw edema in mice. Tropical Journal of Natural Product Research, 7(10), 4152-4159. https://doi.org/10.26538/tjnpr/v7i10.6
Tzanova, M., Atanasov, V., Yaneva, Z., Ivanova, D., & Dinev, T. (2020). Selectivity of current extraction techniques for flavonoids from plant materials. Processes, 8(10), 1222. https://doi.org/10.3390/pr8101222
Wang, M., Li, C., Li, J., Hu, W., Yu, A., Tang, H., Li, J., Kuang, H., & Zhang, H. (2023). Extraction, purification, structural characteristics, biological activity and application of polysaccharides from Portulaca oleracea L. (purslane): A review. Molecules, 28(12), 4813. https://doi.org/10.3390/molecules28124813
Zhang, Y. C., Gan, F. F., Shelar, S. B., Ng, K. Y., & Chew, E. H. (2013). Antioxidant and Nrf2-inducing activities of luteolin, a flavonoid constituent in Ixeris sonchifolia Hance, provide neuroprotective effects against ischemia-induced cellular injury. Food and Chemical Toxicology, 59, 272-280. https://doi.org/10.1016/j.fct.2013.05.058
