Gravity Column Chromatography Research Articles

Acorus calamus L. rhizome extract and its bioactive fraction exhibits antibacterial effect by modulating membrane permeability and fatty acid composition.

Ethnopharmacological relevanceIndia's ancient texts, the Charak Samhita and Sushruta Samhita, make reference to the traditional medicinal usage of Acorus calamus L. In India and China, it has long been used to cure stomach aches, cuts, diarrhea, and skin conditions. This ability of the rhizome is attributed to its antimicrobial properties. Research studies to date have shown its antimicrobial properties. However, scientific evidence on its mode of action is still lacking. Aim of the studyAcorus calamus L. rhizome extract and its bioactive fraction exhibits antibacterial effect by modulating membrane permeability and fatty acid composition. Material and methodThe secondary metabolites in the rhizome of A. calamus L. were extracted in hexane using Soxhlet apparatus. The ability of the extract to inhibit multidrug resistant bacterial isolates, namely Bacillus cereus, Escherichia coli, Acinetobacter baumannii, and Pseudomonas aeruginosa were evaluated using checkerboard assay. Further, the extract was purified using thin layer chromatography, gravity column chromatography, and combiflash chromatography. Structure elucidation of the active compound was done using GC-MS, FT-IR, and UV–Vis spectral scan. The mode of action of the bioactive fraction was determined. Bacterial membrane damage was analyzed using SEM, membrane permeability was determined using SYBR green I and PI dye, leakage of cytoplasmic contents were analyzed using Bradford assay and Fehling’s reagent. The ability to inhibit efflux pump of A. baumannii was determined using EtBr accumulation assay and β-lactamase inhibition was analyzed using nitrocefin as substrate. Also, the biofilm inhibition of B. cereus was determined using crystal violet dye. Moreover, the effect of the bioactive fraction on the fatty acid profile of the bacterial membrane was determined by GC-FAME analysis using 37 component FAME mix as standard. ResultsAcorus calamus L. rhizome hexane extract (AC-R-H) demonstrated broad-spectrum antibacterial activity against all the isolates tested. AC-R-H extract also significantly reduced the MIC of ampicillin against all tested bacteria, indicating its bacterial resistance modulating properties. The assay guided purification determined Asarone as the major compound present in the bioactive fraction (S-III-BAF). S-III-BAF was found to reduce the MIC of ampicillin against Escherichia coli (100–25 mg/mL), Pseudomonas aeruginosa (15–3.25 mg/mL), Acinetobacter baumannii (12.5–1.56 mg/ml), and Bacillus cereus (10–1.25 mg/mL). Further, it recorded synergistic activity with ampicillin against B. cereus (FICI = 0.365), P. aeruginosa (FICI = 0.456), and A. baumannii (FICI = 0.245). The mode of action of S-III-BAF can be attributed to its ability to disturb the membrane integrity, enhance membrane permeability, reduce biofilm formation, and possibly alter the fatty acid composition of the bacterial cell membranes. ConclusionThe bioactive fraction of AC-R-H extract containing Asarone as the active compound showed antibacterial activity and synergistic interactions with ampicillin against the tested bacterial isolates. Such activity can be attributed to the modulation of fatty acids present in bacterial membranes, which enhances membrane permeability and causes membrane damage.

Synthesis of silver nanoparticles from coffee parasite leaf extract (Scurrulla ferruginea (Roxb. ex Jack) Danser) and potential activity as an antioxidant and anticancer

This study aim is to know the results of the synthesis of silver nanoparticles from quercetin and Scurrula ferruginea (Roxb. Ex Jack) danser as well as its potential as an antioxidant and anticancer. The solvent used in the maceration extraction method in this research is the polar solvent methanol. Isolation and purification of thick extract compounds from coffee parasite leaves were carried out using liquid vacuum chromatography (CVC) and gravity column chromatography (GCC) methods. Silver (Ag) nanoparticle is made with the stirrer method at a speed of 1500 rpm for 60 minutes at a temperature of 70o C. Silver nanoparticle solution was produced and then characterized using FTIR (Fourier Transform Infra Red) and PSA (Particles Size Analyzer). PSA results analysis using extract coffee parasite size 214 nm particles, whereas using flavonoid isolate quercetin confirmed size 29 nm. FTIR showed that there was a stretching vibration of the OH group at a wave number of 3229.25 cm-1, the C=O function at the wave number 1637.87 cm-1, and the CO functional group at the wave number 1016.31 cm-1. Antioxidant test using the DPPH method was carried out in silver nanoparticle solution using extract Scurrula ferruginea (Roxb. Ex Jack) danser and with quercetin obtained an IC50 value of around 115.4 and 114.9 this result describes as moderate level of antioxidants. Even though the nanoparticle anticancer test was very weak, overall, the synthesis of silver nanoparticles using flavonoid isolates was better than using extracts from coffee parasite leaves.

ISOLATION OF SQUALENE FROM DICHLOROMssETHANE EXTRACT OF CROTON NIGRITANUS LEAF

A precursor of steroids with a variety of biological activities is squalene. The purpose of the study was to identify and describe the industrial/medical chemical found in Croton nigritanus. Using the solvents such as; dichloromethane, ethyl acetate, and n-hexane, the procedure entails a liquid-liquid extraction stage using the partition method and a solid-liquid extraction step using the maceration method. For separation and purification, gravity column chromatography (GCC) and thin-layer chromatography were employed. The isolated chemical's identity was determined using FTIR, 1H NMR and GC-MS spectroscopy. The inquiry successfully extracted and analyzed the major chemical compound which was identified as Squalene.

Isolation, characterization and anti-inflammatory activity of compounds from the Vernonia amygdalina

The need to explore the abundance of natural products cannot be overemphasized particularly in the management of various disease conditions. In traditional medical practice, Vernonia amygdalina has been widely adopted in the management of various inflammatory disorders. The objective of this investigation was to isolate the bioactive principles from the stem-bark and root of V. amygdalina and assess the anti-inflammatory (in vitro) activity of both the crude extracts and the isolated compounds. Following extraction with the methanol, the extract was subjected to gravity column chromatography and the resultant fractions was further purified to obtained pure compounds. The structural elucidation of the compounds were based on data obtained from 1H to 13C nuclear magnetic resonance (NMR) spectroscopies as well as fourier transform infrared (FT-IR). Using diclofenac as a control drug, the albumin denaturation assay was used to determine the in vitro anti-inflammatory activity of the extracts and isolates. Three distinct compounds characterized are vernoamyoside D, luteolin-7-α-o-glucuronide, and vernotolaside, a new glycoside. When compared to diclofenac, which has an IC50 of 167.8 μg/mL, luteolin-7-α-o-glucuronide, vernoamyoside D, and vernotolaside all showed significant inhibitions with respective IC50 values 549.8, 379.5, and 201.7 μg/mL. Vernotolaside is reported for the first time from the root. The assertion that the plant is used in traditional medicine for the management of inflammatory disorder is somewhat validated by the confirmation of the existence of the compounds with the biochemical actions. Further validation of the isolated compounds would be required in animal studies.

Isolation and Identificationof Octadecane-3-on Compound From Ethyl Acetate Fraction of Momordica balsamina L. Leaves

Momordica balsamina L. is a wild plant that generally grows in moist soil and gets a lot of sunlight. The leaves and fruit are reported to have various medicinal and nutritional properties. This study aimed to isolate and identify the compounds contained in the ethyl acetate fraction in Momordica balsamina L The isolation stage was carried out by the maceration method and the separation and purification of the compounds using gravity column chromatography methods to obtain isolate fraction F1.6.2 with a sample weight of 2.4 mg. Identification of compounds using proton nuclear magnetic resonance (1H-NMR) and carbon nuclear magnetic resonance (13C-NMR) spectroscopy techniques. The proposed identified compound is octadecan-3-one or 3-octadecanone with the molecular formula C18H36O

Isolation of New Asiatic Acid derivative from Centella asiatica (L.) Urban as Antioxidant

Centella asiatica (L.) Urban is known to have a pharmacological effect as an antioxidant. Indonesian research on gotu kola, especially the ethyl acetate fraction it produces, is rarely carried out. Previous research stated that kaempferol and quercetin which isolated from the ethyl acetate fraction of Centella asiatica herb had antioxidant activity with respective IC50 values of 16.50 and 21.61 μg/mL, so this study aimed to get other compounds from the ethyl acetate fraction which has antioxidant activity. The purposes of this study were to isolate, identify, and examine the free radical scavenging activity of DPPH from the ethyl acetate fraction of gotu kola (C.asiatica (L.) Urban) herb. Isolation of the ethyl acetate fraction was carried out from 70% ethanol extract of Centella asiatica plant using vacuum liquid chromatography and gravity column chromatography afforded a new asiatic acid derivative. Elucidation of a new compound based on UV-Vis, FT-IR, LC-MS/MS, 1D-NMR and 2D-NMR to get 11-hydroxy-9-(hydroxymethyl)-1,2,6a,6b,9,12a-hexamethyl-10-((3,4,5-trihydroxytetrahydrofuran-2-yl)oxy )-1,2,3,4,4a,5,6,6a,6b,7,8,8a,9,10,11,12,12a,12b,13,14b-icosahydropicene-4a-carboxylic acid and it had moderate antioxidant activity with IC50 of 106.99 ± 1.76 µg/mL. The pure isolate obtained is suspected to be a new compound derived from asiatic acid.

ISOLATION AND STRUCTURE ELUCIDATION OF SECONDARY METABOLITE COMPOUNDS FROM Curcuma aeruginosa

Black turmeric (Curcuma aeruginosa) is Indonesian medicinal plant belonging to the Zingeberaceae family, mostly found on Java. This study aimed to determine the new molecular structures of the compounds isolated from black turmeric originating from Indonesia. Isolation of the secondary metabolite compounds was initiated by solid-liquid extraction using the maceration method with ethanol solvent, followed by liquid-liquid extraction using the partition method with n-hexane and dichloromethane as solvents. Separation and purification were carried out using vacuum liquid chromatography (VLC) and gravity column chromatography (GCC). The isolated compounds were characterized using spectroscopic methods, including UV-Vis, IR, and NMR spectroscopy. The isolation resulted in three sesquiterpenoid compounds, (E)-7-isopropyl-4,10-dimethylcyclodec-10-ene-5,8-dione (curdione), (1R,10R)-Epoxy-(-)-1,10-dihydrocurdione, and zedoalactone B, together with two common terpenes, ethyl tetradecanoate and 1-hexadecene.

Formation Reaction of 17α(chlorovinyl-3-methyl-butene)estradiol Through the Addition Reaction of a Terminal Alkyne to Ethinylestradiol With a ZnCl2 Catalyst

The addition reaction had been done on the terminal alkynes of 17α-ethynylestradiol with ZnCl2 and HCl catalyst mediated by dichloromethane. The reaction conditions were carried out at a temperature of 70°C for 29 h, the separation of the reaction properties was carried out using Gravity Column Chromatography, and the reaction products were identification using by Nuclear Magnetic Resonance (NMR). The catalyst used was a cationic ligand coordinate, where Zn acts as the central ion and 2-propanol as the ligand. In the reaction mechanism, the reaction begins with the formation of dimeric ether. ZnCl3 catalyst acts as Lewis acid which converts alcohol to dimeric ether and forms a complex with ZnCl3. The nucleophilic terminal alkyne then attacks the C-O bond of the ether to form a vinyl carbocation which then attracts a chlorine atom from ZnCl3 which was complex bound with ether, resulting in a reaction product of 17α(chlorovinyl-3-methyl-butene)estradiol

Synthesis of Cardanol-Based Acetaldehyde Novolac Resin from Cashew Nut Shell Liquid (CNSL)

Polymers are currently in the limelight. Phenolic resin is one of the polymer products obtained from the polymerization process, either from natural or synthetic sources. One of the natural compounds that can be applied to polymers is cardanol. This study aimed to synthesize cardanol-based acetaldehyde novolac resin from Cashew Nut Shell Liquid (CNSL). Isolation of cardanol from CNSL was done by liquid-liquid extraction method with acetone and purified by gravity column chromatography in a mixture of n-hexane and ethyl acetate in a ratio of 9:1. HPLC analysis showed that cardanol compounds had aliphatic chains with different levels of saturation and the yield was 18.48%. The cardanol-acetaldehyde novolac resin was synthesized through a condensation polymerization reaction with a cardanol-acetaldehyde mole ratio (1:0.5), using HCl as a catalyst. FTIR and 1H-NMR analysis were employed to identify the novolac resin structure. The product was a brownish-orange solid with a yield of 12.80 mg (40%) and followed ortho-ortho substitution. Cardanol is one of the natural phenol sources that might be utilized to manufacture novolac resins.

Potential antioxidants of secondary metabolite isolates ethyl acetate fraction Coleus amboinicus Lour. Leaves.

The aim of the study was to isolate and characterize secondary metabolites that have the potential as antioxidants from the ethyl acetate fraction of the leaves of Coleus amboinicus, L. (C. amboinicus).
 Materials and methods. Purification of the ethyl acetate fraction of C. amboinicus using gravity column chromatography with a stationary phase (silica gel, Merck) and a mobile phase with a solvent ratio of n-hexane (Merck) and ethyl acetate (Merck). Examining isolates includes physical (colour, shape, and melting point). Qualitative purity detection by TLC at 254 nm and 366 nm wavelengths. Structural analysis of metabolites with UV-Vis spectrometer (Spectronic 3000, Genesis 10, Japan), FT-IR(KBr) (Shimadzu IR Prestige-21, Japan), NMR spectrometer (JEOL spectrometer, Japan) operating at 500 MHz (1H-NMR) and 125 MHz (13C-NMR), and Shimadzu's GC-MS (QP-2010S Shimadzu, Japan) and determination of potential antioxidant activity using the DPPH method.
 Results. The secondary metabolite compounds were isolated in the form of yellow crystals with a melting point of 232-233 °C and Rf values of 0.86 and 0.56, which TLC monitored at a solvent ratio of n-hexane and ethyl acetate 6:4 and 8:2. Spectronic analysis with a UV-Vis Spectrometer showed two electron absorbances, namely a wavelength of 210 nm indicating methanol solvent and 272 nm isolate. The absorbance of functional groups at wave numbers 3379 cm-1 (-OH; hydroxy), 2931 cm-1 (-CH; aliphatic), 1735 cm-1 (-C=O; carbonyl ketone), 1234 cm-1 (-CO-; methoxy) and 1643 cm-1 (-C=C-; alkene). GC-MS analysis obtained two absorbance peaks, (1) the first retention time of 6.658 minutes (3.95 %) and (2) the second retention time of 9.001 minutes (96.05 %). Structural analysis with 1H&13C-NMR showed 28 types of protons and 22 types of carbon. The antioxidant activity potential test showed an activity value (IC50) of 338.54 ppm.
 Conclusion. The structure of the isolated secondary metabolite compound is 16-acetoxy-7α-hydroxyroyleanone (syn. 16-acetoxyhorminone) and has the potential as an antioxidant.

Isolation, In Vitro and In Silico Anti-Alzheimer and Anti-Inflammatory Studies on Phytosteroids from Aerial Parts of Fragaria × ananassa Duch.

Based on the pharmacological importance of different species of fragaria, this research was carried out for the isolation of bioactive compounds from Fragaria × ananassa. Using the conventional gravity column chromatography followed by small analytical column purification, two major components were isolated from the plant materials. The structures of both compounds (1 and 2) were accurately confirmed with GC-MS analysis by comparison of the fragmentation pattern within the library of the instrument. Further, the NMR analysis was also used to supplement the structural evidence. Compound 1 was observed to be 4,22-cholestadien-3-one, while compound 2 was identified as stigmast-4-en-3-one. Both compounds were evaluated for anticholinesterase, COX/LOX inhibitions and antioxidant assays. Compound 1 exhibited the IC50 values of 20.29, 27.35, 10.70, 80.10 and 7.40 μg/mL against acetylcholinesterase, butyrylcholinesterase, COX-2, COX-1 and 5-LOX, respectively. Similarly, the IC50 values of compound 2 against the same targets were 14.51, 10.65, 8.45, 109.40 and 8.71 μg/mL. Similarly, both compounds were less potent in ABTS and DPPH targets with IC50 values in the range of 185.83–369.86 μg/mL. Despite the low potencies of these compounds in antioxidant targets, they can be considered as supplementary targets in Alzheimer and inflammation. The molecular docking studies for the in vitro anti-Alzheimer and anti-inflammatory targets were also performed, which showed excellent binding interactions with the respective target proteins. In conclusion, the isolated phytosteroids from Fragaria × ananassa were evaluated scientifically for anti-Alzheimer and anti-inflammatory activities using in vitro and molecular docking approaches.

ISOLASI DAN KARAKTERISASI SENYAWA FENOLIK DARI DAUN PUTAT (Planchonia valida Blume) (ISOLATION AND CHARACTERIZATION OF PHENOLIC COMPOUND FROM PUTAT LEAVES (Planchonia valida Blume))

Gallic acid have been isolated from the leaves of the Putat plant (Planchonia valida Blume). This study aimed to isolate and characterize isolates from Putat leaves by phytochemical and FTIR spectrophotometry. The research procedures included extraction by maceration, partitioning by fractionation, vacuum liquid chromatography (KCV), gravity column chromatography (KKG), thin layer chromatography (TLC), and FTIR analysis. A total of 1400 grams of putat leaf powder was macerated to produce 159.243 grams (11.38%) of methanol extract. A total of 101 grams of methanol extract was dechlorophyllated to obtain 21.430 grams (21.21%) of dechlorophyllated extract. The dechlorophyllated extract was partitioned gradually with n-hexane and ethyl acetate to produce n-hexane fractions of 0.341 grams (1.591%), and ethyl acetate of 10.143 grams (47.33%), and methanol of 9.170 grams (42.79%). A phytochemical test showed that in the dechlorophyllated extract, the methanol and ethyl acetate fractions contained alkaloids, phenolics, flavonoids, and terpenoids while the n-hexane fraction contained alkaloids and flavonoids. A total of 7 grams of ethyl acetate fraction was fractionated by vacuum liquid chromatography to produce fractions B1 to B8. The B4 fraction was continued to the separation step by gravity column chromatography so that the fractions S1 to S9 were obtained. The S4 fraction was continued to the separation stage using preparative thin layer chromatography (TLC) to produce isolates S4.1 to S4.5. Isolate S4.1 was tested for purity by two-dimensional TLC showed that the isolate was not pure. The results of the phytochemical test with 5% FeCl3 spray reagent indicated a positive phenolic compound which was strengthened by FTIR characterization of the presence of –OH, -CH aromatic, -CH aliphatic, C=C, C=O and C-O goups. Based on these results, it was concluded that the isolate from the dechlorophyllated extract of Putat leaves belonged to phenolic compounds.

Anti-cervical Cancer Test of the Ethyl Acetate Fraction of Terminalia catappa Linn Fruit Flesh against HeLa Cells

The ethyl acetate fraction from methanol extract of ketapang (Terminalia catappa Linn) fruit flesh has anti-cancer potential. Therefore, this study aims to obtain the active anti-cervical cancer agent from the ethyl acetate fraction of ketapang fruit flesh’s methanol extract. The first step carried out was fractionation of the ethyl acetate fraction of methanol extract using gravity column chromatography with an eluent of ethyl acetate: chloroform: glacial acetic acid (1:8:1). The second step was a toxicity test on fractions produced from column chromatography. The third step was the test of secondary metabolite and anti-cancer activity of fraction possessing the highest toxicity to Artemia salina Leach. Fractionation of the concentrated ethyl acetate fraction by column chromatography, yielded seven subfractions (F1-F7). Furthermore, the cytotoxic test on A. salinaLeach shrimp larvae generated the following LC50 data from the 1st to 7th subfraction in a row, namely 566.2814 ppm, 87.9077 ppm, 216.6232 ppm, 566.2814 ppm, 560, 6647 ppm, 279.9213 ppm, and 194.6674 ppm. The most active subfraction is fraction 2 which have two groups of positive compounds, i.e. phenolics and saponins were obtained from the secondary metabolite test. Data from the infrared spectroscopy performed showed the presence of groups –OH, C-H aromatic, C=O carbonyl, C=C aromatic, C=C alkene, C-H aliphatic, C-H alkane, and C-O. The value of the anti-cancer activity of fraction 2 was IC50 = 165.37 ppm, which was included in the fairly active category as an anti-cervical cancer agent.

Bioassay-guided fractionation and characterization of neuroprotective compounds from the flowers of Achillea millefolium

Oxidative stress is known to play a vital role in the progression of neurodegenerative diseases. The current treatment method primarily targets symptoms by using anti-Parkinson drugs such as levodopa, carbidopa, dopamine (DA) agonists, monoamine oxidase type B inhibitors, and anticholinergics to replace DA. However, these drugs pose severe side effects when used for a prolonged time. Hence, there is a need for a concerted effort to develop natural compounds that reduce oxidative stress and help in the management of PD symptoms. This study aimed to investigate the neuroprotective effects of the petroleum ether extract of Achillea millefolium and its fractions on SH-SY5Y and SK-N-SH neuroblastoma cells using Bioactivity-guided fractionation. To isolate the bioactive fraction, the crude extract was separated on gravity column chromatography. Of the twenty-one fractions collected, one of the fractions (Fraction 4) was found to be the most potent neuroprotective to neuronal cells exposed to the neurotoxins, rotenone, and 6-hydroxydopamine. The results showed Fraction 4’s ability to reduce apoptosis by scavenging ROS, maintaining mitochondrial membrane potential, increasing superoxide dismutase activity, and increasing Sirtuin 3 presence and activity. Fraction 4 also showed the ability to reduce cellular apoptosis through the regulation of the AKT cell signaling pathway. The major compounds present in Fraction 4 were analyzed on TOF-LC-MS. Phosphatidic acid (PA 32:0) and four diacylglycerols (DAG) compounds were identified and characterized for their neuroprotective effect.

AKTIVITAS ANTIOKSIDAN DAN KARAKTERISASI ISOLAT DARI FRAKSI n-HEKSANA PADA RUMPUT LAUT (Caulerpa sertularioides) ASAL SINGKAWANG KALIMANTAN BARAT (ANTIOXIDANT ACTIVITY AND ISOLATE CHARACTERIZATION OF n-HEXANE FRACTION IN SEAWEED (Caulerpa sertularioides) FROM SINGKAWANG WEST KALIMANTAN)

Seaweed (Caulerpa sertularioides) is one of the marine plants of West Kalimantan, which contains bioactive compounds that can act as antioxidants. The purpose of this study was to determine antioxidant activity and identify components of organic compounds in seaweed C. sertularioides. This research was carried out in several stages, namely extraction, fractionation, antioxidant test (DPPH), vacuum liquid chromatography (KVC), gravity column chromatography (KKG), and two-dimensional thin-layer chromatography (2-Dimensional TLC), and characterization. Antioxidant activity test by DPPH method on methanol extract, n-hexane fraction, and fraction DCM from seaweed C. sertularioides obtained IC50 values of 82.21 g/mL (strong antioxidant), 131.2 g/mL (medium antioxidant), and 192.43 g/mL (weak antioxidant). The results of the FT-IR identification of isolate I4 showed a strong absorption at 3483-3387 cm-1,which was specific for the OH group. The presence of strong absorption of 2926-2854cm-1 C-H stretching group, a wide absorption of 1338 cm-1 of methyl and methylene groups (stretching). There was a strong absorption of 1543-1463 cm-1 C-O ester group (bend), a broad absorption of 1074 cm-1 C-O (stretching), and absorption of 1795-1735 cm-1 specific for the C=O ester group. The results of GC-MS on isolate I4 indicated the main component was compound 1-nonadecene (14.09%), Trifluoroacetoxy hexadecane based on the literature, the compound included the organic component of fatty acids, and esters

Antioxidant, Enzyme Inhibitory, and Molecular Docking Approaches to the Antidiabetic Potentials of Bioactive Compounds from Persicaria hydropiper L.

Introduction Natural products are among the most useful sources for the discovery of new drugs against various diseases. Keeping in view the ethnobotanical relevance ethnopharmacological significance of Polygonaceae family in diabetes, the current study was designed to isolate pure compounds from Persicaria hydropiper L. leaves and evaluate their in vitro and in silico antidiabetic potentials. Methods Six compounds were isolated from the chloroform-ethyl acetate fractions using gravity column chromatography and were subjected to structure elucidation process. Structures were confirmed using 1H-NMR, 13C-NMR, and mass spectrometry techniques. Isolated phytochemicals were subjected to in vitro antidiabetic studies, including α-glucosidase, α-amylase inhibition, and DPPH, and ABTS antioxidant studies. Furthermore, the in silico binding mode of these compounds in the target enzymes was elucidated via MOE-Dock software. Results The isolated compounds revealed concentration-dependent inhibitions against α-glucosidase enzyme. Ph-1 and Ph-2 were most potent with 81.84 and 78.79% enzyme inhibitions at 1000 µg·mL−1, respectively. Ph-1 and Ph-2 exhibited IC50s of 85 and 170 µg·mL−1 correspondingly. Likewise, test compounds showed considerable α-amylase inhibitions with Ph-1 and Ph-2 being the most potent. Tested compounds exhibited considerable antioxidant potentials in both DPPH and ABTS assays. Molecular simulation studies also revealed top-ranked confirmations for the majority of the compounds in the target enzymes. Highest observed potent compound was Ph-1 with docking score of −12.4286 and formed eight hydrogen bonds and three H-pi linkages with the Asp 68, Phe 157, Phe 177, Asn 241, Glu 276, His 279, Phe 300, Glu 304, Ser 308, Pro 309, Phe 310, Asp 349, and Arg 439 residues of α-glucosidase binding packets. Asp 68, Glu 276, Asp 349, and Arg 439 formed polar bonds with the 3-ethyl-2-methylpentane moiety of the ligand. Conclusions The isolated compounds exhibited considerable antioxidant and inhibitory potentials against vital enzymes implicated in T2DM. The docking scores of the compounds revealed that they exhibit affinity for binding with target ligands. The enzyme inhibition and antioxidant potential of the compounds might contribute to the hypoglycemic effects of the plant and need further studies.

POTENTIAL OF ACTIVE COMPOUNDS LEAVES OF MANGO PARASITE (Dendrophthoe pentandra (L). Miq) AS ANTIFUNGAL

Infection is a disease that is easily found in tropical areas such as Indonesia. The causes of infection that are easily found include infections due to fungi. One of the fungi that causes many infections is the Candida fungus. Tropical drugs commonly used to treat cutaneous candidiasis include nystatin, clotrimazole, miconazole, ketoconazole and other azoles. However, antifungal drugs have limitations, such as severe side effects, poor penetration into certain tissues, and the emergence of resistance. To overcome the negative effects caused by these synthetic antifungal drugs, it is necessary to explore natural antifungal drugs. One of the plants that has the potential as a medicinal plant is the mango parasite plant (Dendrophthoe pentandra (L.) Miq) which is a parasitic plant that attaches to the host which has the potential as herbal medicine. Research on the potential of active compounds in the leaves of mango parasite (Dendrophthoe pentandra (L). Miq) as antifungals has been carried out from November 2020 to June 2021 at the Genetics and Biotechnology Laboratory, Biology Department, FMIPA Sriwijaya University. The purpose of this study was to obtain fractions that had antifungal activity, to obtain isolates that had antifungal activity, what active compounds were present in the leaves of the mango parasite as antifungal, and to determine the minimum inhibitory concentration of the fungus Candida albicans. Based on research that has been carried out with methanol, n-hexane and ethyl acetate extracts that are active as antifungals, namely n-hexane and ethyl acetate extracts with an n-hexane inhibition zone diameter of 12 mm while ethyl acetate is 10 mm. Then purification of the n-hexane and ethyl acetate fractions was carried out using gravity column chromatography and the n-hexane fraction obtained two pure compounds that were active as antifungals, namely isolate N-1 with a diameter of 12 mm and N-4 with a diameter of 10 mm. Two pure acetate fractions were obtained which were active as antifungals, namely isolates E-1 with a diameter of 10 mm and E 5 with a diameter of 12 mm. Furthermore, the results of the eluate were tested by MIC and thin layer chromatography (TLC). The MIC results on isolates N-1, N-4, and E-1 were able to inhibit at a concentration of 500 ppm and were classified as strong. Meanwhile, isolate E-5 was able to inhibit at a concentration of 1000 ppm which was classified as moderate. The TLC results showed color spots on isolate N-1 which were blue, namely terpenoids, while in isolate N-4 which was orange in color were alkaloids, while in isolate E-1, the color was phenolic yellow, while in isolate E-5, which was purple, it was group terpenoids. The results of the bioautography of isolate N-1 with R¦ value: 0.9, isolate N-4 with R¦: 0.6, isolate E-1 with R¦ value: 0.8, and isolate E-5 with R¦ value: 0,9.

Characterization and Anticancer Activity from Gaharu (Aquilaria malaccensis) Stem Bark Extract

Aquilaria malaccensis is a plant species from the Thymeleaceae family. A. malaccensis contains secondary metabolites of phenolic groups, namely benzophenone, chromone, coumarin, xanthone, and flavonoids with various biological activities such as antioxidants and anticancer. This study aims to determine the potential of A. malaccensis stem bark extract as an anticancer and to determine the characteristics of secondary metabolite compounds contained in A. malaccensis stem bark extract. Extraction of the stem bark of A. malaccensis by maceration method in stages using n -hexane, ethyl acetate and methanol as solvents. The anticancer activity test used the microculture tetrazolium (MTT) assay against Murine Leukemia P-388 cells. Compound separation using thin layer chromatograph (TLC) and gravity column chromatography (CC) methods. Determination of chemical compounds using liquid chromatography mass spectroscopy/mass spectroscopy (LCMS/MS). The extracts of n -hexane, ethyl acetate, and methanol each had anticancer activity which was classified as active with an IC 50 value of 20.64, 26.61, and 14.49 µg/mL. Based on the results of separation and identification of compounds in methanol extract with TLC, CC, and LCMS/MS, methanol extract contains acetaminophen, aminosamptothesin, isophorone, diethyl phthalate, 3-hydroxy-6H-benzochromen-6-one, isolongifolenic, 2-(2-phenethyl) chromone, 4′,7-dihydroxy-2′,5-dimethoxyisoflavone, 4-hydroxy-3-(1-phenylpropyl)-2H-chromone-2-one; 6-methylchromone-2-carboxylic acid; 3,4-dimethyl-2- phenylmorpholine; and β-calacoren compounds.

Antibacterial activity of secondary metabolites isolated from Ficus aurata (Miq.) fruits

This research aimed to isolate chemical constituents from ethyl acetate and n-hexane fractions of Ficus aurata (Miq.) fruits and investigated their antibacterial activities. Chemical constituents were separated by using vacuum column chromatography, gravity column chromatography, and thin layer chromatography. The structures of the isolated compounds 3,4-dihydroxybenzoic acid (1) and β-sitosterol (2) were elucidated using various spectroscopic methods, including UV, IR, and NMR (1H-NMR, 13C-NMR, and DEPT). While, antibacterial activity was done by assay using Resazurin Microtiter Assay method. These 2 compounds reported as new constituents from F. aurata fruit. In addition, analysis of the bioassay showed that compound 1 had a notable antibacterial activity against gram negative (Escherichia coli ATCC 25922 and Pseudomonas aeruginosa ATTC 27853), and gram-positive bacteria (Staphylococcus aureusATCC 23235) with the Minimum Inhibitory Concentration (MIC) value of 0.1563 µg/mL, and it indicated the same MIC as cefadroxil (positive control). Compound 2 has weak antibacterial activity compared to cefadroxil (positive control). The results found that Ficus aurata (Miq.) fruits metabolites have antibacterial activity and show potency as antibacterial agent. Different kinds of bacteria and assay could be needed to investigate and support its antibacterial activity in the future.

Antiproliferative Activity of Triterpenoid and Steroid Compounds from Ethyl Acetate Extract of Calotropis gigantea Root Bark against P388 Murine Leukemia Cell Lines

Calotropis gigantea has been known to produce bioactive secondary metabolites with antiproliferative activities against cancer cells. Herein, we extracted the secondary metabolites using ethyl acetate from its root bark and further tested its antiproliferative activities against P388 murine leukemia cell lines. The subfractions from the ethyl acetate extract was obtained from Vacuum Liquid Column Chromatography (VLCC), and followed by Gravity Column Chromatography (GCC). The subfraction C2 and D1 were identified to contain triterpenoids and steroids with the most potent cytotoxicity against Brine Shrimp Lethality Test (BSLT). A 3-(4,5-dimethylthiazol-2-yl) -2-5 diphenyl tetrazolium bromide (MTT) assay suggested that ethyl acetate extract has the highest antiproliferative activities against P388 murine leukemia cell lines (IC50 = 21.79 μg/mL), as opposed to subfraction C2 (IC50 = 50.64 µg/mL) and subfraction D1 (IC50 = 49.33 µg/mL). The compound identified in subfraction C2 and D1 are taraxerol acetate and calotropone, respectively. Though taraxerol acetate and calotropone were active in inhibiting the leukemic cell lines, their IC50s were lower than the ethyl acetate extract, which is probably due to the synergism of the secondary metabolites.