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Extraction of Gnetum gnemon L. (Melinjo) Leaves Using Deep Eutectic and Conventional Solvents: Phytochemical Contents, Antioxidant Capacity, and Toxicity Assessment
(2026-02-01)
Rushdy, Muhammad Naufal Qaweim
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Awang, Mohd Azrie
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Benjamin, Mohammad Amil Zulhilmi
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Saini, Aniza
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Mohd Rosdan, Muhammad Daniel Eazzat
Gnetum gnemon L. (Melinjo) is widely recognised for its rich bioactive compounds with potential health benefits. This study evaluates the effectiveness of deep eutectic solvents (DESs) and conventional solvents using ultrasound-assisted extraction on the extraction efficiency, antioxidant activity, and toxicity of G. gnemon leaf extracts (GGLE). Four DESs, namely choline chloride-lactic acid (ChCl-LA), choline chloride-glycerol (ChCl-Gly), betaine-lactic acid (Bet-LA), and L-proline-citric acid (LP-CA), were compared with conventional solvents, namely water, methanol, and ethanol. Extraction performance was evaluated based on crude extract yield (CEY), phytochemical content through chlorogenic acid content (CAC), total phenolic (TP), and total flavonoid (TF), as well as antioxidant activity assessed using 2,2-diphenyl-1-picrylhydrazyl (DPPH), 2,2’-azino-bis(3-ethylbenzothiazoline-6-sulfonic acid) (ABTS), and ferric reducing antioxidant power (FRAP) assays. Toxicity was assessed using the brine shrimp lethality assay (BSLA). Among all solvents, ChCl-LA yielded the highest CEY (20.23 ± 0.68%), CAC (13.96 ± 0.31 µg/g), TP (92.54 ± 0.74 mg GAE/g), DPPH (84.95 ± 1.49%), ABTS (91.25 ± 0.34%), and FRAP (481.14 ± 2.31 mg TE/g) (p < 0.05), while exhibiting no toxicity with a high LC₅₀ value (6,339.43 µg/mL). Nonetheless, ethanol yielded the highest TF (2.98 ± 0.47 mg QE/g), followed by ChCl-Gly (2.45 ± 0.27 mg QE/g), although non-significant (p > 0.05), with ChCl-LA (2.14 ± 0.27 mg QE/g) placed as the third highest. Overall, DESs, particularly ChCl-LA, demonstrated promising extraction efficiency and non-toxicity compared to conventional solvents, supporting their potential as greener alternatives for extracting bioactive compounds from G. gnemon leaves.
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Discovery and characterization of bioactive compounds from Limnophila aromatica: nevadensin and related flavonoids as potent antimicrobial agents
(2025-08-01)
Maswanna, Thanaporn
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Maneeruttanarungroj, Cherdsak
Limnophila aromatica, a traditional medicinal plant, has been previously reported to possess notable antimicrobial properties. However, the specific bioactive constituents responsible for this activity remain largely unidentified. This study aimed to isolate, identify, and evaluate the antibacterial potential of compounds from a 100% ethanolic extract of L. aromatica. The crude ethanolic extract exhibited the highest antibacterial activity against Methicillin-resistant Staphylococcus aureus (MRSA) and was subsequently subjected to Diaion HP-20 column chromatography, followed by preparative HPLC. Seven major peaks were identified using UV-Vis spectra, LC-QTOF-MS, and NMR analyses. The antibacterial efficacy of these isolated compounds was assessed using disc diffusion and broth microdilution assays against Bacillus subtilis, S. aureus, MRSA, Escherichia coli, and Pseudomonas aeruginosa. The isolated compounds were identified as norethindrone acetate, isothymusin, nevadensin, gardenin B, 5,3’-dihydroxy-7,8,2’-trimethoxyisoflavone, jasmolin II, and oleanolic acid. Nevadensin, the predominant compound (79.55%), demonstrated potent bactericidal activity against B. subtilis, S. aureus, and MRSA. Jasmolin II and gardenin B also exhibited promising antibacterial effects. Although the disc diffusion assay was limited by compound diffusion, the broth microdilution method confirmed significant MIC values, ranging from 0.59 to 2.86 mg/ml while the MBC values ranged from 0.59 to 12.72 mg/ml. This study highlights nevadensin and other flavonoids from L. aromatica as potential candidates for the development of alternative antibacterial therapies, particularly against drug-resistant Gram-positive bacteria.
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Paclobutrazol enhances yield and secondary compound accumulation in Curcuma longa L.
(2024-11-01)
Montri, N.
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Chauytam, S.
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Deewatthanawong, R.
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Bunya-Atichart, K.
The results indicated that the application of different concentrations of PBZ at 60 days preharvest had an impact on yield after harvest at 9 months while the color of turmeric remained relatively consistent. In all treatments, the rhizome color was yellow to orange. The fresh weight of the rhizome showed no significant differences within the range of 0-800 mg/L PBZ treatments, with the lowest yield observed in 1,200 mg/L PBZ treatment. The dry weight percentage was highest when treated with 800 mg/L PBZ for the mother rhizome and 600 mg/L PBZ for the finger rhizome. Regarding the accumulation of secondary compounds, highly significant differences among the treatments were found for the content of total flavonoids and total phenolics while curcumin showed significant differences. The treatment with 1,200 mg/L PBZ had the highest total phenolics content at 78.18 mg GAE/gDW and total flavonoids at 295.54 mg QUE/gDW. PBZ-treated plants also showed higher curcumin content and percentage than the non-treated plants. These findings indicated that the application of PBZ has the potential to improve both the yield and quality of turmeric, which is advantageous for cultivating this valuable medicinal plant.
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Antioxidant, Anti-Oral Cancer, and Antimicrobial Activity of Medicinal Plant Extracts: Development of Mouthrinse Formulations
(2023-04-01)
Nanasombat, Suree
Nowadays, there is growing interest in using medicinal plants as antimicrobial agents to treat microbial infections in the oral cavity. This study was aimed to evaluate phytochemical properties, anti-oral cancer and antimicrobial activity against C. albicans and some potential oral lactic acid bacteria (Lactobacillus casei and Lactobacillus plantarum). Plant extracts including Cinnamomum verum (cinnamon), Acorus calamus (sweet flag), Hibiscus sabdariffa (roselle), Zingiber officinale (ginger) and Alpinia officinarum (lesser galangal) were tested and used to formulate mouthrinses for oral health care. Cinnamon extract possessed the highest phenolic, flavonoid and tannin content, and antioxidant activity (1,152.58 mg ascorbic acid equivalent/g by phosphomolybdenum assay and 39.64 % nitric oxide radical scavenging activity), while roselle extract had the highest total alkaloids. Only lesser galangal extract displayed anti-oral cancer activity with 97.83 % cytotoxicity. High performance liquid chromatography (HPLC) analysis revealed the presence of galangin and kaempferide in the lesser galangal rhizome extract. The cinnamon extract showed the strongest inhibitory activity against Candida albicans (4 mg/mL MIC). Cinnamon extract (24 mg/mL) and cinnamaldehyde (0.08 mol/L) exhibited the best killing effect and the highest cellular protein leakage in C. albicans. In addition, these plant extracts were used to develop mouthrinse formulations. The extracts of cinnamon and ginger at 10 % in mouthrinse formulations could strongly inhibit the growth of C. albicans. In comparison, 8 and 10 % roselle extracts showed the best inhibitory effect against Lactobacillus casei. In summary, cinnamon and lesser galangal were potential plants with antimicrobial, antioxidant, anti-biofilm and anti-oral cancer activities, and thus can be utilized to formulate oral care products.
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In vitro fermentation characteristics and methane mitigation responded to flavonoid extract levels from alternanthera sissoo and dietary ratios
(2021-09-01)
Sommai, Sukruthai
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Cherdthong, Anusorn
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Suntara, Chanon
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So, Sarong
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Wanapat, Metha
Two experiments were conducted under this study: Experiment 1 was to study production yield, chemical composition, and in vitro degradability of Brazilian spinach (Alternanthera sissoo; BS) leaf and leaf + leaf-stalk at various maturity ages of 15, 30, 45, and 60 days after plantation and regrowth and Experiment 2 was to evaluate the effect of flavonoid extract from BS leaf and leaf + leaf-stalk and dietary ratios on ruminal gas production, fermentation characteristics, and in vitro degradability. Experiment 1 showed that maturity ages after planting and regrowth increased, the yield significantly increased. Increasing maturity ages significantly (p < 0.05) increased neutral detergent fiber and acid detergent fiber content and decreased crude protein content, total flavonoid (TF) content, and degradability for both leaf and leaf + leaf-stalk. Maturity ages from 15 to 30 days after plantation and regrowth resulted (p < 0.05) the highest TF content and degradability for both leaf and leaf + leaf-stalk. Thus, BS leaf and leaf + leaf-stalk samples from 15 to 30 days of age were used for flavonoid extraction and used in the Experiment 2. Experiment 2 was conducted according to a 3 × 5 factorial experiment. Three roughage to concentrate (R:C) ratios at 50:50, 40:60, and 30:70 were used, and five levels of flavonoid extract (FE) at 0, 10, 20, 30, and 40 mg of substrate dry matter (DM) were supplemented. Experiment 2 showed that R:C ratio and FE had an interaction effect only on acetate to propionate ratio. Varying R:C ratios significantly increased (p < 0.05) in vitro DM degradability, total volatile fatty acids (VFA), and propionate (C3) concentration. FE supplementation linearly (p < 0.05) increased total VFA and C3 concentration and decreased methane production and protozoal population. This study could conclude that FE from BS could effectively modulate ruminal fermentation and decrease methane production. However, in vivo study needs to elucidate in order to validate the present results.
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Enhancements of growth and metabolites of indica rice callus (Oryza sativa l. cv. pathumthani1) using tio2 nanoparticles (nano-tio2)
(2020-04-01)
Chutipaijit, S.
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Sutjaritvorakul, T.
In this research, the effects of a various in concentration of TiO<inf>2</inf> nanoparticles (Nano-TiO<inf>2</inf>) were applied to study the cell growth, metabolites and antioxidant responses of callus in indica rice plants. Dose-dependent changes in growth performances, metabolite accumulations and antioxidant activities of callus were found. Mature seeds of indica rice cv. Pathumthani1 were cultured in callus induction medium with different concentrations of Nano-TiO<inf>2</inf> (0-600 mg L-1). After five weeks of cultivation, the callus formations were measured in fresh weights and dry weights, and the contents of phenolic compounds, flavonoids, and the antioxidant activities of callus extracts were assayed. The Nano-TiO<inf>2</inf> induced of fresh weights and dry weights in callus formations when compared with the control treatment (0 mg L-1 Nano-TiO<inf>2</inf>) were also observed. The flavonoid and phenolic compound contents in rice callus exposed to 400 mg L-1 Nano-TiO<inf>2</inf> were 118% and 139%, respectively, higher than rice callus unexposed to Nano-TiO<inf>2</inf>. Moreover, the antioxidant activities of the callus metabolites studied shows that the percentages of inhibition were up-regulated (84%) when exposed to 400 mg L-1 Nano-TiO<inf>2</inf>, as a response to the high-level depletion of the free radicals in metabolites of rice callus. The results obtained from this research showed that the addition of Nano-TiO<inf>2</inf> had impacts on the callus growth and metabolite accumulations in rice callus. The results suggested that addition of Nano-TiO<inf>2</inf> at appropriate levels could role as an elicitor for biosynthesis of valuable metabolites and antioxidant properties for pharmaceutical applications in further research.
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ZNO AND TIO2 nanoparticles stimulate callus induction and secondary metabolite production in indica RICE
(2020-01-01)
Chutipaijit, Sutee
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Sutjaritvorakul, Thanawat
Metal nanoparticles are widely used in various fields of industry and their impact in the agricultural field is increasing. This research was carried out to investigate the effects of ZnO and TiO<inf>2</inf> nanoparticles on callus induction and plant metabolism in indica rice. Indica rice was assessed in plant tissue culture condition to estimate the influence of metal nanoparticles on morphological and physiological factors over 4 weeks. The seeds of Oryza sativa L. var. KDML105 were exposed to 1-3 mg L-1 of 2,4-dichlorophenoxyacetic acid (2,4-D) in the callus induction medium. The induced-calli were cultured on the callus induction medium supplemented with 2 mg L-1 of 2,4-D and showed a high increase in size and dry weight contents under the light condition when compared with the control and other treatments. Moreover, this research also investigated the potential impacts of ZnO and TiO<inf>2</inf> nanoparticles at 200, 400 and 800 mg L-1 on morphological and metabolic responses of the induced-calli cultured on the callus induction medium supplemented with 2,4-D and each type of metal nanoparticles. At 4 weeks after treatment, the induced-calli were analyzed for plant growth and assays on aspects of their metabolism were undertaken. The induced-calli exposed to 200 mg L-1 of ZnO or TiO<inf>2</inf> nanoparticles were found to have significant increase of biomass (size and dry weight of callus), total phenolic compounds and total flavonoid contents in KDML105 variety. The results demonstrated that utilization of ZnO and TiO<inf>2</inf> nanoparticles at appropriate concentration could positively induce the production of beneficial secondary metabolites in the induced-calli of indica rice and are worthy of further study for applications in other plant species.
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Comparative study of total phenolic compounds, flavonoids and antioxidant capacities in pigmented and non-pigmented rice of indica rice varieties
(2018-06-01)
Chutipaijit, Sutee
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Sutjaritvorakul, Thanawat
The total phenolic compounds and flavonoid contents of grains as well as the antioxidant capacities were determined in 15 rice varieties. A high-performance liquid chromatography method was modified for the determination of the flavonoid subgroups in rice grains. The results showed the total phenolic compounds, flavonoid contents, and antioxidant capacities of pigmented rice were higher than in non-pigmented rice. The amounts of flavanone groups ranging from 0.04 to 0.40 mg Naringenin g−1, while flavones groups ranging from 0.08 to 0.57 mg Apigenin g−1, flavonol groups ranging from 0.16 to 1.20 mg Quercetin g−1 and anthocyanin groups ranging from 44.43 to 69.83 mg Cyanidin chloride g−1 were found in pigmented rice. The total phenolic compounds and total flavonoid contents of grains were significantly correlated with their antioxidant capacities. The results indicated that pigmented rice appeared as a good source of phenolic and flavonoid compounds and had beneficial nutritive values or antioxidant substances.
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Tobacco randomly inserted TT8 differenly enhance light signals and flavonoid accumulation
(2015-08-18)
Sompornpailin, Kanokporn
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Kanthang, Supha
The individual lines of tobacco over-expressing TT8, a bHLH gene, were constructed and cultured under tissue culture condition radiating with photosynthetically activation radiation (PAR) or PAR+UVA. They were compared to wild type (WT). Leaf of treated plants was extracted and analyzed for flavonoid accumulations using a spectrophotometer. The extract of TT8 plants significantly contained flavone, flavonol and anthocyanin level, higher than the WT extract did. The petal extracts of mature transgenic under PAR had a similar absorbance profile of each substance, but these extracts had higher flavonoid contents than the leaf extracts did. All flavonoid subgroups and p-coumaric acid biosynthesis were significantly enhanced after the additional UVA radiation to plant. This UVA condition slightly stimulated an accumulation of these substances in normal plant. Some transgenic greatly increased flavonoid accumulation in responding to PAR+UVA, but the others were slightly different compared to WT. The distinct insertion site is directly affected TT8 gene expression. Transgenic seeds had a dark brown color more than WT seed, which indicated high content of polymer flavonoids (proanthocyanins). This over-expressing TT8 in transgenic tobacco may directly or indirectly enhance the signal transductions of PAR and UVA and raise up flavonoid accumulation.
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Tobacco expressing PAP1 increases the responses to PAR and UV-A by enhancing soluble sugars and flavonoids and elevating plant protections
(2015-04-01)
Sompornpailin, Kanokporn
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Kanthang, Supha
Five lines of transgenic tobacco over-expressing Production of Anthocyanin Pigment 1 (PAP1) cDNA were analysis of metabolic response against the radiation and their protection of the plant under tissue culture condition. PAP1 transgenic and wild type (WT) plants were treated with the radiations of photosynthetically activate radiation (PAR) or PAR combined with UV-A. All lines of transgenic significantly increased in amounts of p-coumaric acid, naringenin apigenin more than WT under both treatments. Additional UV-A radiating to plant rose up kaempferol content in WT plant (1.5 times) and in PAP1 transgenics (1.8 times). These transgenic plants treated under both conditions had also increased anthocyanin substances (pelargonidin) with significant value after compared to WT. Content of total soluble sugar (TSS) was related to the content of total flavonoids in transgenic. PAR combined with UV-A had a lower induction of the electrolyte leakage percentage and malondialdehyde (MDA) level in the transgenic leaf tissue compared to WT tissue. The metabolic substance levels were considered on its protection of plant cells. In transgenic tissue, the enhancement of apigenin level strongly diminished the increase level of electrolyte leakage while the levels of TSS, p-coumaric acid and naringinin less affected. Moreover, the increase levels of kaempferol and pelargonidin associated with the decrease level of MDA, while the TSS level reversely responded. The PAP1 transgenic increased response of light by adaptation of their metabolites (TSS, p-coumaric acid and flavonoids) consequently enhance parameter indicating protections of the cell.

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