Exploration of Total Flavonoid Content, Toxicity, and Antibacterial Activity of Acorus Calamus L. Rhizome Isolates

Background: Andrographis paniculata is a herbaceous plant in the Acanthaceae family, that is widely used as a traditional medicine in Asian countries and known to exhibit a wide range of pharmacological effects. Recent studies have provided an overview of the great potential of A. paniculata as an analgesic. The ethanol extract and ethyl acetate (EA) fraction of A.paniculata were shown to contain diterpene lactone compounds, which may be useful as a potential active ingredient in analgesic drugs. The development of a herbal medicine based drug requires an effective and high quality active ingredient. Therefore, this research was aimed to compare the analgesic activity of ethanol extract and EA fraction based on their andrographolide content and further to determine the more viable active substance for analgesic herbal medicine based drug development. Method: The andrographolide content in the ethanol extract and EA fraction was determined by High Pressure Liquid Chromatography (HPLC). Measurement of analgesic activity was performed by writhing test. The experimental animals were randomly divided into eight groups consisting of 5 mice in each. Group 1 (negative control) received 1% Tween-80 in normal saline. Group 2 (positive control) received a standard analgesic drug (diclofenac sodium) at a dose of 40 mg/kg body weight. Group 3, 4, and 5 received ethanol extract while Group 6, 7, and 8 received EA fraction, each at a dose of 12.5, 25, and 50 mg andrographolide/kg body weight, respectively. Each mouse was injected intraperitoneally with 1% acetic acid at a dose of 10 ml/kg body weight 30 minutes after oral administration of the treatments. The number of writhes were counted 5 min after acetic acid injection over a period of 45 min. Results: Andrographolide content in ethanol extract and EA fraction was 15.66±0.28 and 21.25±1.08 % w/w, respectively. Ethanol extract and EA fraction displayed analgesic activity of 67.68% and 70.91% respectively, at a dose of 50 mg andrographolide/kg body weight. The positive control at a dose of 40 mg/kg body weight showed an analgesic activity of 74.33%. Statistical analysis showed no significant differences between EA fraction at a dose of 50 mg andrographolide/kg body weight and ethanol extract at the same dose as well as the positive control (P> 0.05). The effective dose 50% (ED50) of the ethanol extract and EA fraction was determined to be 29.49 and 25.55 mg/kg body weight, respectively. Conclusion: It was possible to use andrographolide content as an indicator for the analgesic activity of A.paniculata. Ethanol extract and EA fraction of A. paniculata at the same dose of andrographolide showed similar analgesic activity. The amount of ethanol extract which needed to reach similar analgesic activity was higher than EA fraction. Therefore, EA fraction likely has greater potential as an analgesic active substance due to its higher content of andrographolide; however further study is needed to develop it as a dosage form.

This research objectives to determine antioxidant activities and inhibition of α-glucosidase 96 % ethanol extract of gayo arabica coffee skin, n-hexane fraction, ethyl acetate fraction, and water fraction. Gayo arabica coffee skin was extracted by ethanol using maceration and fractionation using n-hexane, ethyl acetate, and water. Quercetin and Gallic acid were used as standard to found total flavonoids and phenolics. Test of antioxidant activity using the DPPH method. After being tested the inhibition of the α-glucosidase IC50 ethanol extract, the n-hexane fraction, ethyl acetate fraction, and water fractions. The total flavonoids of 96% ethanol extract, n-hexane fraction, ethyl acetate, and water fraction respectively are 76.74, 99.07, 73.55, 50.24 mg QE/g extract . Total phenolics of 96% ethanol extract, n-hexane fraction, ethyl acetate, and water fraction respectively are respectively 211.85, 136.67, 145.19, 127.41 mg GAE/ g extract. Antioxidants activity of ethanol extract, n-hexane fraction, ethyl acetate fraction, and water fraction are IC50 12.03 ppm, 36.08 ppm, 30.27 ppm, 11.62 ppm. Inhibition activity α-glucosidase enzyme respectively are 588.85 ppm, 341.81 ppm, 591.61 ppm, and 798.78 ppm. These showed that the ethanol extract and water fraction are the best in antioxidants and the ethanol extract and ethyl acetate fraction are the best in inhibiting the α-glucosidase enzyme. These results indicate that gayo arabica coffee skin extract has the potential to be developed as an antidiabetic compound.
 Keywords: Antidiabetic, Antioxidant, flavonoids, phenolics, α-glucosidase

Diabetes mellitus (DM) is a metabolic disorder characterized by elevated blood glucose levels due to impaired insulin function. Dipeptidyl peptidase 4 (DPP-IV) inhibitors are potential agents for managing type-2 DM. This study evaluates the total phenolic and flavonoid content, antioxidant activity, and DPP-IV inhibitory potential of the ethanolic extract and ethyl acetate fraction of Sargassum cristaefolium. Antioxidant activity was assessed using the DPPH assay, while DPP-IV inhibition was compared with sitagliptin. Gas Chromatography-Mass Spectrometry (GC-MS) was used to identify active compounds. The total phenolic content in the ethanol extract and ethyl acetate fraction was 1.04% and 7.33% GAE, respectively, while total flavonoid content was 1111.70 and 3529.01 µgQE/g. Antioxidant activity increased in a concentration-dependent manner, with IC₅₀ values of 1.38 mg/mL (ethanol extract) and 0.71 mg/mL (ethyl acetate fraction). The ethanolic extract (500 ppm) and ethyl acetate fraction (62.5 ppm) exhibited DPP-IV inhibitory activity of 92.07% and 95.87%, respectively. GC-MS identified hexadecenoic acid, methyl ester, and trans-13-octadecenoic acid, methyl ester as dominant compounds. S. cristaefolium exhibits promising antioxidant and DPP-IV inhibitory activity, supporting its potential role in type-2 DM management.

Avocado (Persea americana Mill.) has long been used in traditional medicine, and its seeds are known to contain flavonoid compounds, which are natural phenolics with antioxidant properties and potential therapeutic benefits. This study aims to identify the chemical compounds present in the ethanol extract and ethyl acetate fraction of avocado seeds, and to determine their total flavonoid content using UV-Visible spectrophotometry. This experimental study included sample collection, simplicia characterization, extraction, fractionation, phytochemical screening, and flavonoid quantification. Phytochemical screening showed that the ethanol extract contains alkaloids, flavonoids, tannins, saponins, triterpenoids, and glycosides, while the ethyl acetate fraction contains alkaloids, flavonoids, tannins, steroids, and glycosides. Flavonoid content was determined using quercetin as a standard. The results showed that the total flavonoid content in the ethanol extract was 8.655 ± 0.144 mgQE/g, while the ethyl acetate fraction contained 32.828 ± 0.105 mgQE/g. It can be concluded that the ethyl acetate fraction contains a significantly higher level of flavonoids compared to the ethanol extract.

Antioxidants are compounds that inhibit free radicals. Many plants have antioxidant activity because they contain antioxidant compounds such as flavonoids. This study aimed to evaluate the antioxidant activity, phytochemical contents, total flavonoid content and to establish the relationship of total flavonoid content and antioxidant activity of extract and fractions of walnut (Canarium indicum L.) leaves. Antioxidant activity of ethanolic extract, n-hexane fraction, ethyl acetate fraction, and ethanolic-water fraction of walnut leaves was examined by using the DPPH method and determination of total flavonoid content was determined spectrophotometrically. The results showed that antioxidant activity of ethanolic extract, n-hexane fraction, ethyl acetate fraction, and ethanolic-water fraction based were 25.294 ± 0.055; 175.245 ± 0.4999; 20.135 ± 0.009; and 28.806 ± 0.0424 μg/ml, respectively. The phytochemical content of ethanol extract of walnut leaves are saponins, flavonoids, tannins, and polyphenols. The total flavonoid content of ethanolic extract, n-hexane fraction, ethyl acetate fraction and ethanolic-water fraction were 2.624 ± 0.012; 0.499 ± 0.023; 3.846 ± 0.006; and 1.596 ± 0.006 gram quercetin equivalent/gram extract, respectively. Correlation between antioxidant activity and total flavonoid content revealed that 63.2 % of antioxidant activity was influenced by the presence of flavonoid compounds.

This study was conducted to investigate the physicochemical properties and antioxidant activity of the solvent fractions obtained from 80% ethanol extracts of Wasabia koreana Nakai leaves. From the ethanol extracts, hexane, chloroform, ethyl acetate, and n-butanol fractions were sequentially extracted and collected, and further used for the investigation. The highest yield was obtained in the water fraction and its pH was 4.25. The total polyphenol, total flavonoid and total pectin contents in the ethyl acetate fraction were 56.24 mgGAE/g, 97.29 mgNE/g and 108.8 mg/g, respectively. This indicated that the extraction yields of the phenolic compounds and pectin in the solvent fractions obtained from the Wasabi leaves and petiole were significantly different. Analysis of the taste components of the ethyl acetate and water fractions was carried out using an electronic tongue sensor. A negative concentration-dependent population for the ethyl acetate fraction and a positive population for the water fraction, corresponding to DF1 (83.96%) in the discriminant function analysis (DFA) plot, were confirmed from the analysis. In the ethyl acetate fraction, the IC50 values ​​of 1,1-diphenyl-2-picrylhydrazyl(DPPH) radical scavenging activity and 2,2’-azino-bis(3-ethylbenzothiazoline-6-sulfonic acid(ABTS) radical scavenging activity were 899.88 ppm and 276.63 ppm, respectively. Therefore, the ethanol extracts and fractions from Wasabia koreana Nakai leaf have low utilization, which will be beneficial for the development of high value-added products via improvement in the taste components and antioxidant activity.

Acorus calamus L. rhizome has potential activity as an antimicrobial agent. This study aims to evaluate the antimicrobial activity of the rhizome extracts and its total flavonoid and phenolic contents. Extraction was carried out by using ethanol at room temperature followed by fractionation into n-hexane, ethyl acetate, and n-butanol. Antimicrobial activity towards Escherichia coli, Staphylococcus aureus, and Candida albicans were evaluated by agar diffusion method. Total flavonoid and phenolic contents were determined by UV-Vis Spectrophotometer using the standard of quercetin and gallic acid, respectively. The A. calamus rhizome ethyl acetate fraction strongly inhibited Escherichia coli, Staphylococcus aureus and Candida albicans, followed by n-butanol and n-hexane fractions, respectively. There was no inhibition of water fraction towards all the microbes. Total flavonoid contents of ethyl acetate, butanol, n-hexane, and water fractions were successively 2978.34, 399.07, 142, 23, and 14.80 mg QE/100g, while total phenolic contents of those fractions were 1820.51, 329.71, 237.81, and 74.36 mg GAE/100g, respectively. The results show that there is a positive correlation between antimicrobial activity and its total flavonoid and phenolic contents.

Jatropha curcas Linn. is a multipurpose plant in the Euphorbiaceae family. Numerous reportshave indicated the antioxidant properties of phenolics and flavonoids present in J. curcas rootmethanolic extract. In the present study, 80% methanolic extract of J. curcas root was preparedand used for extraction of bioactive compounds with five solvents (hexane, chloroform, ethylacetate, n-butanol and water) by liquid-liquid fractionation. The fractions were evaluated fortotal phenolic content (TPC), total flavonoid content (TFC) and antioxidant activities by usingthe 2-2’-diphenyl-1-picrylhydrazyl (DPPH), ferric reducing/antioxidant potential (FRAP) and2,2′-azino-bis-3-ethylbenzthiazoline-6-sulphonic acid (ABTS) assays. The phytochemicalcompounds present in each fraction were identified by liquid chromatography mass spectrometry(LCMS) analysis. The TPC of ethyl acetate and n-butanol fractions were 34.0 ± 0.02 μg GAE/gDW and 33.1± 0.01 μg GAE /g DW, respectively, while the TFC were 9.2 ± 0.04 μg CE/g DWand 10.1 ± 0.01 μg CE/g DW, respectively. The free radical scavenging activity varied with thesolvents used. Ethyl acetate fraction showed the highest inhibition for DPPH (48.7%) and FRAP(79.6%) assays, while n-butanol fraction showed the highest ABTS radical scavenging activity(94.8%). The LCMS analysis showed the compounds present in the various fractions werephenolic and flavonoid derivatives such as coumaric acid, epigenin, quercetin, leuteolin and pcoumaroylquinicacid. The results showed that TPC, TFC and antioxidant activity for ethylacetate and n-butanol fractions were higher compared to the other solvent fractions.

Uncaria nervosa Elmer (local name: Bajakah) is one of the medicinal plants used as a cancer drug by the people of Muara Badak, East Kalimantan. The purpose of this study was to determine the total flavonoid content (TFC), toxicity, and antioxidant activity of the n-hexane (NH) and ethyl acetate (EA) fractions of Uncaria nervosa root wood. Determination of total flavonoids was carried out using a UV-Vis spectrophotometer with quercetin as a standard, toxicity test was carried out using the brine shrimp lethality test (BSLT) method, and antioxidant test using the DPPH radical reduction method. Based on the results of this study, the total flavonoid content (TFC) of NH and EA fractions were 192 and 425 mg QE/g extract, respectively. Based on the results of the toxicity test on Artemia salina L. shrimp, the LC50 values were 11.91 ppm for NH and 33.19 ppm for EA. These LC50 values indicate that NH is very toxic and EA is toxic against Artemia salina L. While the antioxidant test results showed that EA was very significant as an antioxidant and NH was moderate with IC50 values of 17.54 ppm and 75.93, respectively.

Staphylococcus Aureus is a gram-positive bacteria that can cause infection. One of the plants that has antibacterial activity is jatropha leaves which contain flavonoids, saponins, tannins, alkaloids, steroids, and polyphenols. This study aims to determine the antibacterial activity of ethanol extract and ethyl acetate fraction with concentrations of 30%, 60% and 100% against staphylococcus aureus bacteria. The method for extracting jatropha leaves is maceration with 96% ethanol solvent and the fractionation method, namely liquid-liquid fractionation with ethyl acetate solvent. Antibacterial activity test was carried out in vitro with the disc diffusion method and compared the mean zone of inhibition of each treatment with a positive control (gentamicin 10 µg). The results showed that the ethanol extract and ethyl acetate fraction of jatropha leaves had a strong resistance response, while the positive control gave a very strong inhibitory response to the growth of staphylococcus aureu bacteria. Based on the one way ANOVA test, ethanol extract and ethyl acetate fraction showed a significant difference from each treatment (P = ˂0.05). The conclusion of this study is that the ethanol extract of Jatropha leaves can inhibit the growth of staphylococcus aureus bacteria at a concentration of 100% (18.28 ± 0.50mm), 100% concentration of ethyl acetate fraction (15.10 ± 0.12mm). The ethanol extract provided the best inhibition power, namely 18.28 ± 0.50mm and a positive control 21.82 ± 0.092mm

Staphylococcus Aureus is a gram-positive bacteria that can cause infection. One of the plants that has antibacterial activity is jatropha leaves which contain flavonoids, saponins, tannins, alkaloids, steroids, and polyphenols. This study aims to determine the antibacterial activity of ethanol extract and ethyl acetate fraction with concentrations of 30%, 60% and 100% against staphylococcus aureus bacteria. The method for extracting jatropha leaves is maceration with 96% ethanol solvent and the fractionation method, namely liquid-liquid fractionation with ethyl acetate solvent. Antibacterial activity test was carried out in vitro with the disc diffusion method and compared the mean zone of inhibition of each treatment with a positive control (gentamicin 10 µg). The results showed that the ethanol extract and ethyl acetate fraction of jatropha leaves had a strong resistance response, while the positive control gave a very strong inhibitory response to the growth of staphylococcus aureu bacteria. Based on the one way ANOVA test, ethanol extract and ethyl acetate fraction showed a significant difference from each treatment (P = ˂0.05). The conclusion of this study is that the ethanol extract of Jatropha leaves can inhibit the growth of staphylococcus aureus bacteria at a concentration of 100% (18.28 ± 0.50mm), 100% concentration of ethyl acetate fraction (15.10 ± 0.12mm). The ethanol extract provided the best inhibition power, namely 18.28 ± 0.50mm and a positive control 21.82 ± 0.092mm

Background: Staphylococcus aureus is a gram-positive bacteria that can cause infection. One of the plants that has antibacterial activity is jatropha leaves which contain flavonoids, saponins, tannins, alkaloids, steroids and polyphenols. Purpose: To determine the antibacterial activity of ethanol extract and ethyl acetate fraction with concentrations of 30%, 60% and 100% against Staphylococcus aureus bacteria. Method: The method for extracting jatropha leaves is maceration with 96% ethanol solvent and the fractionation method, namely liquid-liquid fractionation with ethyl acetate solvent. Antibacterial activity test was carried out in vitro with the disc diffusion method and compared the mean zone of inhibition of each treatment with a positive control (gentamicin 10 μg). Result: The results showed that the ethanol extract and ethyl acetate fraction of jatropha leaves had a strong resistance response, while the positive control gave a very strong inhibitory response to the growth of S. aureus bacteria. Based on the one way ANOVA test, ethanol extract and ethyl acetate fraction showed a significant difference from each treatment with a significant value (P=<0.05). Conclusion: The ethanol extract of Jatropha leaves can inhibit the growth of S. aureus bacteria at a concentration of 100% (18.28 ± 0.50 mm), 100% concentration of ethyl acetate fraction (15.10 ± 0.12 mm). The ethanol extract provided the best inhibition power, namely 18.28 ± 0.50 mm and a positive control 21.82 ± 0.092 mm.

Diabetes mellitus is a metabolic disease characterized by an uncontrolled increase of blood glucose levels. Diabetes drugs such as metformin, acarbose, and others have the side effects that can harm to the human body such as liver, kidneys and other organs. Tristaniopsis obovata Benn. leaves have efficacy as an α-glucosidase inhibitors (AGIs), but the active compounds contained in T. obovata leaves have not been disclosed. This study aimed to determine the active compound in 70% ethanolic extract of T. obovata leaves which has the potential as AGIs. T. obovata leaves were macerated using 70% ethanol for 3×24 h gave ethanol extract-T. obovata leaves in 22.7% yield. Analysis of total phenolic and flavonoid contents of ethanol extract-T. obovata leaves showed that its total phenolic content was 31.1 mg GAE g-1 and total flavonoid content was 10.9 mg QE g-1 . Solvents partition of ethanol extract gradually gave n-hexane, ethyl acetate, methanol, and water fractions in 10.0, 32.3, 41.0, and 8.9% yields, respectively. In vitro bioassay of extract and its fractions for α-glucosidase enzyme showed that the IC50 values were 3.2, 3.9, 5.1 and 7.5 µg mL-1 for n-hexane fraction, ethyl acetate fraction, methanol fraction and water fraction, respectively, of ethanol extract. Separation of methanol fraction by column chromatograph using silica gel and n-hexane/ethyl acetate/methanol (4/1/0 – 0/1/1) as a mobile phase produced 17 subfractions (F-1 to F-17). In vitro bioassay of F-11, F-15, and F-16 for α-glucosidase enzyme gave the IC50 values of 40.0, 5.0, 3.6 µg mL-1 , respectively. Analysis by FT-NMR spectroscopy, it was predicted that F-16 was myricetin 3-O-α-rhamnoside, which was the first time found in the T. obovata. © 2024 Friends Science Publishers

Staphylococcus aureus and Escherichia coli are among the most common species of gram-positive and gram-negative bacteria, which cause vaginitis, in infertile women. The Calamus rhizome (Acorus calamus L.) is an Indonesian plant that has antibacterial properties that can be used to treat vaginitis and increase fertility. The aim of this study was to determine the phytochemical and antibacterial activity of the calamus rhizoma in polar, semi-polar and non-polar solvents in the growth of S. aureus and E. coli. The antibacterial activity test was in the form of inhibitory test using the Kirby-Bauer, Minimum Inhibi-tion Concentration (MIC) and Minimum Bactericidal Concentration (MBC) by microdilution method with multilevel dilution (concentra-tions 50; 25; 12.5; 6.25; 3.13; 1.56; 0.78; and 0.39%). The screening results showed that ethanol and n-hexane extract contained alkaloids and triterpenoids, while chloroform extract was only triterpenoid. Chloroform extract produced the largest inhibition zone diameter of S. aureus and E. coli (7.26 and 3.28 mm), followed by ethanol extract (5.90 and 3.07 mm) and n-hexane extract (5.33 and 2.95 mm). The concentrations of 0.39 and 0.78% were the values of MIC and MBC for all three extracts, indicating that the extract of the calamus rhizome with several solvents in this study had the same antibacterial activity.

This research was conducted to identify simplicia microscopically, phytochemical screening and determination of total flavonoid content of extract and ethyl acetate fraction from Moringa (Moringa oleifera L.) leaves using UV-Visible Spectrophotometry method. The experimental design used in this study was to perform microscopic identification of Moringa leaf powder simplicia, make 96% and 70% ethanol extract and ethyl acetate fraction of Moringa leaves from 70% ethanol extract, then carry out phytochemical screening and determination of total flavonoid content with quercetin standards. Phytochemical screening on the ethyl acetate fraction of Moringa leaves included tests for the content of flavonoids, saponins, tannins and alkaloids. The results of microscopic identification of Moringa leaf simplicia showed the presence of calcium oxalate crystals in the form of rosettes, mesophyll and stomata. The result of determination of total flavonoid content in 96% ethanol extract was 16.69 ± 0.74% (w/w), 70% ethanol extract was 10.84 ± 0.49% (w/w), Moringa leaf ethyl acetate fraction 14 .45 ± 0.90% (w/w). The highest total flavonoid content was found in the 96% ethanol extract of Moringa leaves in accordance with the 2017 Indonesian Herbal Pharmacopoeia, that the thick extract of Moringa leaves containing no less than 6.30% (w/w) total flavonoids was calculated as quercetin.