skip to main content

The Effect of Various Sterilization Methods and Volume Containers on Phytochemical Content of Methanol Extract of Phyllanthus urinaria

Department of Chemistry, Faculty of Sciences and Mathematics, Diponegoro University, Semarang, Indonesia

Received: 14 Jun 2023; Revised: 19 Oct 2023; Accepted: 2 Nov 2023; Published: 5 Nov 2023.
Open Access Copyright 2023 Jurnal Kimia Sains dan Aplikasi under http://creativecommons.org/licenses/by-sa/4.0.

Citation Format:
Cover Image
Abstract

Phyllanthus urinaria is an annual perennial herbal species found in tropical Asia, America, China, and the Indian Ocean islands. Phyllanthus urinaria is used in folk medicine as a cure to treat jaundice, diabetes, malaria, and liver diseases. Sterilizing the substrate is a crucial step in the fermentation process. This process ensures that the inoculated microorganism is entirely single. Autoclave sterilization is widely favored within the scientific community. In autoclaving, pressurized steam is employed to deliver heat, effectively reducing the bioactive compounds present in the substrate. Comparative studies on various sterilization methods have been reported. This study aims to investigate the effects of substrate containers in sterilization methods of the herbal plant on phenol and flavonoid compounds by LC-MS (Liquid Chromatography-Mass Spectrometry) analysis. Three sterilization methods (pasteurization, steam, and autoclave sterilization) were each applied to the meniran herbal plant (Phyllanthus urinaria) for 15 minutes. Using the aluminum chloride colorimetric assay, the sterilization results were measured for total phenol content, the Folin-Ciocalteu test, and total flavonoid content. The LC-MS analysis showed that the methanol extract of Phyllanthus urinaria (APU) sterilized by autoclaving resulted in the most significant reduction in active phenolic and flavonoid compounds. Pasteurization, steaming, and autoclaving in a big container resulted in total flavonoid content of 1.80±0.034, 1.70±0.021, and 1.71±0.029 mg QE/g extract. The total phenolic content was 26.49±0.591, 22.77±0.230, and 22.097±0.155 mg GAE extract/g, respectively. Meanwhile, using a small container, each method produced a total flavonoid content of 1.73±0.024, 1.71±0.051, and 1.62±0.015 mg QE/g extract, respectively. The total phenolic content was 20.56±0.093, 19.79±0.295, and 20.09±0.124 mg GAE/g extract. Furthermore, the LC-MS profile revealed that APU experienced a reduction in ρ-hydroxybenzaldehyde and naringenin compounds, leading to a decrease in rutin, methyl brevifolincarboxylate, and ethyl gallate compounds. From the results of LC-MS analysis, this research determined that pasteurization using a big container is the most effective sterilization method for preserving the highest levels of total flavonoid and phenolic content in Phyllanthus urinaria while minimizing adverse effects on phytochemical compounds.

Fulltext View|Download
Keywords: herbal extract; phytochemical compounds; sterilization method; Phyllanthus urinaria; LC-MS

Article Metrics:

  1. Madamanchi Geethangili, Shih-Torng Ding, A Review of the Phytochemistry and Pharmacology of Phyllanthus urinaria L., Frontiers in Pharmacology, 9, (2018), 1109 https://doi.org/10.3389/fphar.2018.01109
  2. Chun Wu, Chun-Shan Wei, Shao-Fu Yu, Bai-Lian Liu, Yao-Lan Li, Wen-Cai Ye, Guang-Dong Tong*, Guang-Xiong Zhou*, Two new acetylated flavonoid glycosides from Phyllanthus urinaria, Journal of Asian Natural Products Research, 15, 7, (2013), 703-707 https://doi.org/10.1080/10286020.2013.794792
  3. Dian Wahyu Harjanti, Fajar Wahyono, Vincentia Rizke Ciptaningtyas, Effects of different sterilization methods of herbal formula on phytochemical compounds and antibacterial activity against mastitis-causing bacteria, Veterinary World, 13, 6, (2020), 1187-1192 www.doi.org/10.14202/vetworld.2020.1187-1192
  4. Mercedes M. Pedrosa, Eva Guillamón, Claudia Arribas, Autoclaved and Extruded Legumes as a Source of Bioactive Phytochemicals: A Review, Foods, 10, 2, (2021), 379 https://doi.org/10.3390/foods10020379
  5. Diksha Gupta, Neelam Chaturvedi, A Comparative Study on Impact of Blanching and Autoclaving on Nutraceutical Profile of Helianthus Tuberosus L. (Jerusalem Artichoke), Carpathian Journal of Food Science & Technology, 13, 2, (2021), 43-53 https://doi.org/10.34302/crpjfst/2021.13.2.4
  6. Paula Ribeiro Buarque Feitosa, Tacila Rayane Jericó Santos, Nayjara Carvalho Gualberto, Narendra Narain, Luciana Cristina Lins de Aquino Santana, Solid-state fermentation with Aspergillus niger for the bio-enrichment of bioactive compounds in Moringa oleifera (moringa) leaves, Biocatalysis and Agricultural Biotechnology, 27, (2020), 101709 https://doi.org/10.1016/j.bcab.2020.101709
  7. Sunil Kumar, Awantika Singh, Brijesh Kumar, Identification and characterization of phenolics and terpenoids from ethanolic extracts of Phyllanthus species by HPLC-ESI-QTOF-MS/MS, Journal of Pharmaceutical Analysis, 7, 4, (2017), 214-222 https://doi.org/10.1016/j.jpha.2017.01.005
  8. N. A. Ibrahim, S. Mustafa, A. Ismail, Effect of lactic fermentation on the antioxidant capacity of Malaysian herbal teas, International Food Research Journal, 21, 4, (2014), 1483-1488
  9. Aminah Aminah, Nurhayati Tomayahu, Zainal Abidin, Penetapan kadar flavonoid total ekstrak etanol kulit buah alpukat (Persea americana Mill.) dengan metode spektrofotometri UV-Vis, Jurnal Fitofarmaka Indonesia, 4, 2, (2017), 226-230 https://doi.org/10.33096/jffi.v4i2.265
  10. Ramadan ElGamal, Cheng Song, Ahmed M. Rayan, Chuanping Liu, Salim Al-Rejaie, Gamal ElMasry, Thermal Degradation of Bioactive Compounds during Drying Process of Horticultural and Agronomic Products: A Comprehensive Overview, Agronomy, 13, 6, (2023), 1580 https://doi.org/10.3390/agronomy13061580
  11. Laila Ayuni Hidayah, Mirwa Adipraha Anggarani, Determination of Total Phenolic, Total Flavonoid, and Antioxidant Activity of India Onion Extract, Indonesian Journal of Chemical Science, 11, 2, (2022), 123-135 https://doi.org/10.15294/ijcs.v11i2.54610
  12. Amjad M. Shraim, Talaat A. Ahmed, Md Mizanur Rahman, Yousef M. Hijji, Determination of total flavonoid content by aluminum chloride assay: A critical evaluation, LWT, 150, (2021), 111932 https://doi.org/10.1016/j.lwt.2021.111932
  13. L. G. Malta, R. H. Liu, Analyses of Total Phenolics, Total Flavonoids, and Total Antioxidant Activities in Foods and Dietary Supplements, in: N.K. Van Alfen (Ed.) Encyclopedia of Agriculture and Food Systems, Academic Press, Oxford, 2014, https://doi.org/10.1016/B978-0-444-52512-3.00058-9
  14. Miguel Palma, Zulema Piñeiro, Carmelo G. Barroso, Stability of phenolic compounds during extraction with superheated solvents, Journal of Chromatography A, 921, 2, (2001), 169-174 https://doi.org/10.1016/S0021-9673(01)00882-2
  15. Kunlestiowati Hadiningrum, Ratu Fenny Muldani, Optimization of the amount of gas moles determination through Boyle’s law and Gay-Lussac’s law experiments, 2018, 2, 02, (2018), 11 https://doi.org/10.20961/jphystheor-appl.v2i2.30666
  16. Panin Poolchak, Wittaya Kanchanapusakit, An equipment design to verify Boyle’s law, Journal of Physics: Conference Series, 1144, (2018), 012075 https://doi.org/10.1088/1742-6596/1144/1/012075
  17. Y. Martono, F. F. Yanuarsih, N. R. Aminu, J. Muninggar, Fractionation and determination of phenolic and flavonoid compound from Moringa oleifera leaves, Journal of Physics: Conference Series, 1307, (2019), 012014 https://doi.org/10.1088/1742-6596/1307/1/012014
  18. José R. Vergara-Salinas, Jara Pérez-Jiménez, Josep Lluís Torres, Eduardo Agosin, José R. Pérez-Correa, Effects of Temperature and Time on Polyphenolic Content and Antioxidant Activity in the Pressurized Hot Water Extraction of Deodorized Thyme (Thymus vulgaris), Journal of Agricultural and Food Chemistry, 60, 44, (2012), 10920-10929 https://doi.org/10.1021/jf3027759
  19. Vellingiri Maheshu, Deivamarudhachalam Teepica Priyadarsini, Jagathala Mahalingam Sasikumar, Effects of processing conditions on the stability of polyphenolic contents and antioxidant capacity of Dolichos lablab L., Journal of Food Science and Technology, 50, (2013), 731-738 https://doi.org/10.1007/s13197-011-0387-z
  20. Ahmed Olatunde, Aminu Mohammed, Mohammed Auwal Ibrahim, Mohammed Nasir Shuaibu, Influence of methoxylation on the anti-diabetic activity of ρ-hydroxybenzaldehyde in type 2 diabetic rat model, Phytomedicine Plus, 1, 1, (2021), 100003 https://doi.org/10.1016/j.phyplu.2020.100003
  21. Bradley W. Bolling, Almond Polyphenols: Methods of Analysis, Contribution to Food Quality, and Health Promotion, Comprehensive Reviews in Food Science and Food Safety, 16, 3, (2017), 346-368 https://doi.org/10.1111/1541-4337.12260
  22. Guankui Du, Man Xiao, Siman Yu, Mengyi Wang, Yiqiang Xie, Shenggang Sang, Phyllanthus urinaria: a potential phytopharmacological source of natural medicine, International Journal of Clinical and Experimental Medicine, 11, 7, (2018), 6509-6520
  23. Min Xu, Zhong-Jun Zha, Xue-Ling Qin, Xiang-Lan Zhang, Chong-Ren Yang, Ying-Jun Zhang, Phenolic Antioxidants from the Whole Plant of Phyllanthus urinaria, Chemistry & Biodiversity, 4, 9, (2007), 2246-2252 https://doi.org/10.1002/cbdv.200790183
  24. Li-jun Yao, Jian-qing Wang, Hong Zhao, Jian-she Liu, An-guo Deng, Effect of telmisartan on expression of protein kinase C-α in kidneys of diabetic mice, Acta Pharmacologica Sinica, 28, 6, (2007), 829-838 https://doi.org/10.1111/j.1745-7254.2007.00541.x
  25. Zhengxi Hu, Yongji Lai, Jinwen Zhang, Ye Wu, Zengwei Luo, Guangmin Yao, Yongbo Xue, Yonghui Zhang, Phytochemical and chemotaxonomic studies on Phyllanthus urinaria, Biochemical Systematics and Ecology, 56, (2014), 60-64 https://doi.org/10.1016/j.bse.2014.04.016
  26. Shih-Hua Fang, Yerra Koteswara Rao, Yew-Min Tzeng, Anti-oxidant and inflammatory mediator's growth inhibitory effects of compounds isolated from Phyllanthus urinaria, Journal of Ethnopharmacology, 116, 2, (2008), 333-340 https://doi.org/10.1016/j.jep.2007.11.040
  27. Adair R. S. Santos, Rafael O. P. De Campos, Obdúlio G. Miguel, Valdir Cechinel-Filho, Rosendo A. Yunes, João B. Calixto, The involvement of K+ channels and Gi/o protein in the antinociceptive action of the gallic acid ethyl ester, European Journal of Pharmacology, 379, 1, (1999), 7-17 https://doi.org/10.1016/S0014-2999(99)00490-2
  28. Assieh Behdad, Sasan Mohsenzadeh, Majid Azizi, Comparison of phytochemical compounds of two Glycyrrhiza glabra L. populations and their relationship with the ecological factors, Acta Physiologiae Plantarum, 42, (2020), 133 https://doi.org/10.1007/s11738-020-03121-0
  29. Matej Ravber, Darja Pečar, Andreja Goršek, Jernej Iskra, Željko Knez, Mojca Škerget, Hydrothermal Degradation of Rutin: Identification of Degradation Products and Kinetics Study, Journal of Agricultural and Food Chemistry, 64, 48, (2016), 9196-9202 https://doi.org/10.1021/acs.jafc.6b03191
  30. Mercedes M. Pedrosa, Carmen Cuadrado, Carmen Burbano, Mercedes Muzquiz, Blanca Cabellos, Begoña Olmedilla-Alonso, Carmen Asensio-Vegas, Effects of industrial canning on the proximate composition, bioactive compounds contents and nutritional profile of two Spanish common dry beans (Phaseolus vulgaris L.), Food Chemistry, 166, (2015), 68-75 https://doi.org/10.1016/j.foodchem.2014.05.158
  31. Moonkyeung Choi, Yu-Ra Kang, Hyo Don Zu, In-Sook Lim, Sung Keun Jung, Yoon Hyuk Chang, Effects of Time on Phenolics and in vitro Bioactivity in Autoclave Extraction of Graviola (Annona muricata) Leaf, Biotechnology and Bioprocess Engineering, 25, (2020), 9-15 https://doi.org/10.1007/s12257-019-0259-3
  32. David Gregg, John N. Saddler, A techno-economic assessment of the pretreatment and fractionation steps of a biomass-to-ethanol process, Applied Biochemistry and Biotechnology, 57, (1996), 711-727 https://doi.org/10.1007/BF02941753
  33. H.-H. Goh, K. Khairudin, N. A. Sukiran, M. N. Normah, S. N. Baharum, Metabolite profiling reveals temperature effects on the VOCs and flavonoids of different plant populations, Plant Biology, 18, S1, (2016), 130-139 https://doi.org/10.1111/plb.12403

Last update:

No citation recorded.

Last update: 2024-11-21 08:38:13

No citation recorded.