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FR-07

Evaluation on chemical composition, anti-oxidative, and anti-inflammatory

activity of Marsilea minuta L.

1
1
1
1
Prangruthai Boorapha, Jutiporn Nundee, Kittiyakan Prasopsin, Benyapha Srinop,
2
Tanavin Sobhon, Yi-Pei Li, Sheng-Ting Lin, Ching-Yun Liu, Y u -Chia Chang, Ming-
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4,5
1
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2,6
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Yi Lee, 2,3,4 Ching-I Lin, Passakorn Kingwascharapong,* Cheng-Pei Chung* ,2,3,6

1 Department of Fishery Products, Kasetsart University, Bangkok 10900, Thailand
2 Department of Nutrition and Health Sciences, Chang Gung University of Science and
Technology, Taoyuan 333324, Taiwan
3 Graduate Institute of Health Industry Technology, Chang Gung University of Science and
Technology, Taoyuan 3333234, Taiwan
4 Center for Drug Research and Development, Chang Gung University of Science and
Technology, Taoyuan 333324, Taiwan
5 Department of Cosmetic Science, Chang Gung University of Science and Technology,
Taoyuan 333324, Taiwan
6 Research Center for Food and Cosmetic Safety, College of Human Ecology, Chang Gung
University of Science and Technology, Taoyuan 333324, Taiwan
* E-mail: passakorn.ki@ku.th; cpchung@mail.cgust.edu.tw

Abstract
Marsilea minuta L. (MM), a perennial aquatic fern native to Taiwan, has historically been
consumed as food and used for treating malaria, neurasthenia, hepatitis, and edema. In folk
medicine, the whole plant is applied, whereas medicinal uses of local varieties abroad typically
involve only the stems and leaves. According to previous reports, many of the biological
activities of MM are associated with inflammatory responses. In the present study, the whole
plant of MM was extracted with ethanol to obtain ethanolic extract (MME), which was
subsequently partitioned into n-hexane (MME-Hex), ethyl acetate (MME-EA), 1-butanol
(MME-BuOH), and water (MME-H₂O) fractions. Total polyphenol contents (TPCs) and
flavonoid contents were measured, and the anti-oxidative and anti-inflammatory activities were
evaluated using 2,2-diphenyl-1-picrylhydrazyl (DPPH) free radical scavenging assay, ferric
reducing antioxidant power (FRAP), and lipopolysaccharide (LPS)-induced nitric oxide (NO)
production in RAW 264.7 macrophages. The results showed that MME-BuOH and MME-H2O
contained relatively high levels of TPCs, whereas MME-EA exhibited the highest flavonoid
content. MME-EA and MME-BuOH demonstrated better DPPH-scavenging activities of 31.0
± 0.0 and 33.5 ± 0.2% at 500 µg/mL as compared with other fractions, while all fractions
displayed limited FRAP activity. MME at 25 µg/mL in combination with LPS affected RAW
264.7 cell viability, but none of the fractions influenced viability up to 50 µg/mL. Notably, all
fractions significantly suppressed LPS-induced NO production in dose-dependant manner,
approaching half-maximal inhibitory levels at 50 µg/mL. In conclusion, MM exhibits
promising anti-inflammatory potential, warranting further investigation into its chemical
constituents and underlying mechanisms.

Keywords: Marsilea minuta L.; DPPH free radical; RAW 264.7 macrophages; NO production