Vacuum Liquid Chromatography Research Articles

Isolation and characterization of bioactive phytochemicals from chloroform extract of Adenodolichos paniculatus (hua) Hutch. & Dalz (Fabaceae)

Phytochemicals are the chemicals extracted from plants which make them significant source of drugs with potential for thousands of years. Adenodolichos paniculatus have been observed to have some medicinal applications. The aim of this work was to isolate and characterize some bioactive phytochemicals from the chloroform root extract of A. paniculatus which may justify its use by traditional healers. A. paniculatus roots were collected, identified, dried and pulverized. The pulverized plant material was subjected to serial exhaustive extraction using n-hexane, chloroform, ethyl acetate, methanol and water. Vacuum liquid chromatography of the chloroform root extract led to a number of fractions. TLC analysis was used to study the different fractions. This isolation and purification afforded a dark orange liquid which was subjected to physical spectroscopic identification using IR and GC-MS. The major phytochemicals identified were 9,12-octadecadienoic acid (linoleic acid), undecanoic acid, 10-bromo-(10-bromoudecanoic acid), octadecanoic acid (stearic acid) and n-hexadecanoic acid (palmitic acid). These findings support the traditional use of Adenodolichos paniculatus in various diseases treatments specifically sore throat infections.

Immunomodulatory Activity of Phyllanthus maderaspatensis in LPS-Stimulated Mouse Macrophage RAW 264.7 Cells

Phyllanthus species (Family Euphorbiaceae) has been used in traditional medicine of several countries as a cure for numerous diseases, including jaundice and hepatitis. This study is an attempt to evaluate the immunomodulatory activity of various fractions, column eluents of ethyl acetate fraction, and their polyphenols. Phyllanthus maderaspatensis were standardized using high-performance liquid chromatography to identify and quantify polyphenols, and purification of polyphenols was carried out using vacuum liquid chromatography. Subsequently, we tested various fractions, column eluents of ethyl acetate fraction, and polyphenols in vitro to assess their impact on nitric oxide (NO) production in LPS-stimulated mouse macrophage RAW 264.7 cells. The ethyl acetate fraction (100 μg mL−1) had a more significant stimulatory effect on LPS-stimulated NO production by the RAW 264.7 cells. We found that the ethyl acetate fraction contains a high amount of catechin, quercetin, ellagic acid kaempferol, and rutin, which are responsible for immunomodulation. The ethyl acetate fraction at concentrations of 25 and 50 μg mL−1 had a significant inhibitory effect and 100 μg mL−1 had a more significant stimulatory effect when compared with the LPS control. The percentage of inhibition by LPS control ranged from zero percentage, kaempferol ranged from 45.4% at 50 μg mL−1 to 41.88% at 100 μg mL−1, catechin ranged from 50% at 50 μg mL−1 to 35.28% at 100 μg mL−1, rutin ranged from 36.2% at 50 μg mL−1 to 47.44% at 100 μg mL−1, gallic acid ranged from 28.4% at 50 μg mL−1 to 50.9% at 100 μg mL−1, ellagic acid ranged from 45.12% at 50 μg mL−1 to 38.64% at 100 μg mL−1, and purified quercetin ranged from 26.2% at 50 μg mL−1to 45.48% at 100 μg mL−1. As NO plays an important role in the immune function, polyphenols’ treatment could modulate several aspects of host defense mechanisms owing to the stimulation of the inducible nitric oxide synthase.

Isolation, peroxisome proliferator-activated receptor-gamma transcription, glucose uptake, and molecular docking of tetramethoxyflavonoids from the leaves of Rourea mimosoides (Vahl) Planch.

Rourea mimosoides (Vahl) Planch. is used traditionally to treat various diseases. The ethanolic leaves extract was found to increase the transcription of peroxisome proliferator-activated receptor-gamma (PPARγ). Therefore, the present study is warranted to identify the active compounds from the dichloromethane and ethanolic leaves extract and examine their PPARγ binding and glucose uptake activity. The ground dried leaves were macerated successively in dichloromethane and ethanol and dried and fractionated using vacuum liquid chromatography. Exhaustive chromatographic procedures yielded to a new tetramethoxyflavonoid, quercetin tetramethyl ether-3-O-�-L-rhamnoside (1), along with a known quercetin tetramethyl ether (2) and a steroid (3). Compound 2 showed the highest PPARγ binding and glucose uptake activities at 0.313 and 2.5 �g/ml, respectively. The leaves extract and compounds 1�3 showed no cytotoxicity when evaluated using HepG2 cells. Molecular docking result showed that compound 2 is able to interact with important residues within the hydrophobic pocket of the ligand-binding domain, thus acting as an agonist of PPARγ.

Screening for anti-inflammatory activity of extract and fractions of Morinda lucida stem bark and detection of bioactive secondary metabolite

Morinda lucida Benth (Rubiaceae) is a versatile plant used in traditional medicine for the treatment of a variety of ailments and the claims of its efficacy are particularly remarkable in the treatment of infections and immuno-inflammatory disorders. This study evaluates the anti-inflammatory properties of methanolic stem bark extract and fractions of M. lucida and also identifies the phytochemicals responsible for its anti-inflammatory activity. The crude extract was subjected to liquid- liquid partitioning successively with n- hexane, ethylacetate, butanol and water. High performance liquid chromatography (HPLC) of the four fractions and Vacuum Liquid Chromatography fraction (VLC) of the promising fraction was evaluated. The effect of the fractions on egg albumen induced rat paw oedema were also evaluated. Anti-inflammatory activity of the fractions was further screened using xylene induce ear oedema models and human red blood cell membrane stabilization test. Ulcerogenic test on the normal stomach mucosa was also evaluated. The result of the egg albumen induced rat paw oedema showed that the butanol fractions maximally inhibited egg albumen induced effect at 400 mg/kg (70%) and 200mg/kg (67.5%) after 180 minutes compared to the positive control, ibuprofen (20mg/kg) with 100% inhibition after 180 minutes. The result of the xylene induced ear oedema showed that the inhibition produced by 100 µg/ear of the Butanol fraction (BF) was 56.67 % and was greater than inhibition produced by 200 µg/ear of ibuprofen (38.89 %). HPLC analysis of the fractions revealed the following phytocompounds; Cytreo- a-pyrone, Cytosporin- J and Waol A. Ulcerogenic test was negative at the doses of 200 mg/kg and 400 mg/kg of the fractions when compare with the indomethacin (positive control) at dose of 50 mg/kg. Human red blood cell membrane stabilization assay showed that BF-VLC 2 (Dichloromethane: methanol (8:2) VLC of Butanol fraction) exhibited concentration dependent inhibition of heat-induced haemolysis while other extract showed a non- concentration dependent inhibition of haemolysis when compared to the standard, ibuprofen. These findings suggest that the stem bark of M. lucida possess promising anti-inflammatory phytocompounds which justify its use in ethno-medicine.

Two Limonoids from The Seeds of Chisocheton Macrophyllus and Their Cytotoxic Activity Against Mcf-7 Breast Cancer Cells

Limonoids (tetranortriterpenoids) are triterpenoid compounds that lose four terminals in their structural framework. These compounds have a wide variety of structures and interesting activities including anti-inflammatory, anti-cancer, anti-tumor and anti-malarial properties. The purpose of this study was to find limonoid compounds from the Indonesian Chisocheton plant, and one of which is Chisocheton macrophyllus. The dried and powdered seeds of C. macrophyllus (4.5 kg) were extracted with methanol and partitioned successively with n-hexane, ethyl acetate and n-butanol. Evaporation of each extract resulted in the crude extracts of n-hexane (346.6 g), ethyl acetate (60.8 g) and n-butanol (14.6 g). The n-hexane fraction was subjected to a silica gel vacuum-liquid chromatography (VLC) column packed with silica gel 60 using gradient of n-hexane, ethyl acetate and methanol (10% stepwise) to afford thirteen fractions (A-M). Fraction F (4.4 g) was subjected to silica gel column chromatography using gradient of n-hexane and ethyl acetate (5% stepwise). Subfraction F5 (1.2 g) was chromatographed on a column of silica gel eluted with n-hexane: CH2Cl2: EtOAc (2:7.5:0.5) to give compound 1 (19.7 mg) and fraction H (1.8 g) was subjected to silica gel column chromatography using gradient of n-hexane and ethyl acetate (5% stepwise) as eluting solvent to give 2 (12.0 mg). Chemical structures of 1 and 2 were elucidated by spectroscopic methods and determined as 6α-acetoxyepoxyazadiradione (1) and Dysobinin(2). Dysobinin (2) showed weak cytotoxic activity against MCF-7 breast cancer cells with an IC50 value of 228.15 μM

Antimicrobial Evaluation of Extracts and Fractions of Daniella oliveri Leaf on Selected Clinical Enteropathogens

Background: Diarrhoea is one of the major health threats to the populace in the tropics, and also one of the killer diseases in children under 5 years of age. Antimicrobial resistance and its spread pose serious public health threats, hence the need for development of safer and more effective antibacterial agents. Daniella oliveri Hutch and Dalz is used in ethnomedicine for the treatment of diarrhoea and other gastrointestinal disturbances.
 Objective: To investigate the antimicrobial activity of Daniella oliveri leaves on diarrhoeal pathogens.
 Methods: Successive plant extraction was carried out with hexane, ethylacetate and methanol using soxhlet apparatus. Methanol extract was fractionated using vacuum liquid chromatography (VLC). Phytochemical screening was done using standard chemical assays. Antibiogram of test isolates was carried out using disc diffusion assay. Antimicrobial activity of plant extract and fractions against strains of diarrhoeagenic Escherichia coli, Salmonella cholerae-suis, Shigella dysenteriae, Proteus mirabilis and Acinetobacter baumannii was determined by agar well diffusion and MIC by agar dilution methods. Kill-kinetics study was carried out using viable count technique.
 Results: Terpenoids, steroids and anthraquinones were detected in fractions of D. oliveri. Antibiogram assay showed that 66% of isolates were MDR. Extract and fractions produced appreciable zones of inhibition on all challenge organisms except Acinetobacter baumannii. MICs ranged between 6.25-25 mg/ml. Kill kinetics studies showed total kill on susceptible pathogens after 24 hours.
 Conclusion: This research has shown D. oliveri is a promising drug candidate for the management and treatment of diarrhoea.

NMR Study of Methyl β-orsellinate from Dirinaria sp. was Collected from Coastal Areas of Teluk Nipah, Pulau Pangkor, Perak Malaysia

The research was carried out to isolate and characterize the methyl β-orsellinate of lichen Dirinaria sp. collected from the coastal areas of Teluk Nipah, Pulau Pangkor, Perak Malaysia. The sample was extracted in methanol for three times at room temperature. Subsequently, the crude extracts and further analysed using thin-layer chromatography (TLC) and vacuum liquid chromatography (VLC). To derive the pure compounds, the isolation step was performed using radial chromatography (RC). The chemical structure of the isolated compound was determined by several spectroscopies i.e. infra-red (IR), nuclear magnetic resonances (NMR), and mass spectrometric (MS). The compound was identified as methyl β-orsellinate.

Acetylcholinesterase inhibitory activity of extract and fractions from the root of Rauvolfia serpentina(L.) Bth.ex Kurz.

Alzheimer's disease (AD) is a degenerative brain disease characterized by confusion, behavior changes, decline in memory and cognitive skills. One of the strategies in the treatment of AD is to use acetylcholinesterase (AChE) inhibitors. The current study aims to determine the AChE inhibitory activities of the extract and fractions of the root of Rauvolfia serpentina. Extraction was carried out by maceration method using ethanol, followed by liquid-liquid partition using n-hexane, ethyl acetate and n-butanol. Further fractionation was conducted by using vacuum liquid chromatography (VLC). The AChE inhibitory assays were performed by using Ellmann's method. Phytochemical screening was carried out by TLC method. The ethanolic extract of R.serpentina showed inhibition against AChE enzyme with an IC50 value of 7.46μg/mL. The extract and fractions showed higher inhibition against butyrylcholinesterase (BChE) compared to AChE. Amongst three fractions obtained, the n-butanol fraction showed the strongest inhibition with an IC50 value of 5.99μg/mL against AChE. VLC fractionation of the n-butanol fraction yielded 13 subfractions (VLC 1-VLC 13). Four out of 13 subfractions gave more than 80% inhibition against AChE, namely subfractions 4-7, with IC50 values ranging from 4.87 to 47.22μg/mL. The phytochemical screening of these subfractions suggested the presence of alkaloids. The ethanolic extract, as well as fractions of R.serpentina root, are potential for AChE inhibitor. The alkaloid compound may be responsible for this activity.

Identifikasi Senyawa Terpenoid dan Steroid pada Beberapa Tanaman Menggunakan Pelarut N-Heksan

Hexane is a hydrocarbon compound of alkanes with the chemical formula C6H14. The prefix hex refers to the six carbon atoms found in the hexane and the suffix ana is derived from the word alkane which refers to the single bond connecting the carbon atoms. Identification of terpenoid compounds and steroids has been identified in several plants, namely Red Shoots (Syzygium myrtifolium Walp.), Salam (Syzygium polyanthum), bilaran Tapah (Argyreia nervosa (Burm. F.), Blind (Excoecaria agallocha L.), as well as on the lotion preparations Citrus hystrix DC. This study was conducted to determine the content of terpenoid compounds and steroids found in some parts of plants using n-hexane solvents. Research used is qualitative research with Lieberman Burchard method. Review this article using a method of literature study by searching sources and literature with data form in the form of national journals to international journals about the last 10 years (2012-2021). Identification is carried out on extracts, fractions, isolates and lotion preparations using liquid vacuum chromatography, column chromatography, thin-layer chromatography, and Lieberman-Burchard reagents. The results showed that all of the samples were positive for terpenoid compounds and steroids using n-hexane solvents.

Ingol and Ingenol-Type Diterpenes from Euphorbia trigona Miller with Keratinocyte Inhibitory Activity.

Ingenol mebutate, isolated from Euphorbia peplus, is an ingenane-type diterpenoid, primarily used for the topical treatment of actinic keratosis, a premalignant skin condition. The aim of our work was to investigate other Euphorbia species to find structurally similar diterpenes that can be used as alternatives to ingenol mebutate. Pharmacological investigation of Euphorbia candelabrum, Euphorbia cotinifolia, Euphorbia ramipressa, and Euphorbia trigona revealed the potent keratinocyte (HPV-Ker cell line) inhibitory activity of these spurge species. From the methanolic extract of the aerial parts of Euphorbia trigona Miller, the most active species, five ingol (1–5) and four ingenane-type diterpenoids (6–9) were isolated by various chromatographic separation techniques, including open column chromatography, vacuum liquid chromatography, thin-layer chromatography, and high-performance liquid chromatography. The structures of the compounds were determined by NMR spectroscopic analysis and by comparison of the assignations with the literature data. The cytotoxic activity of the compounds against keratinocytes was tested in vitro by using ingenol mebutate as a positive control. Among the isolated compounds, two ingenane derivatives (6 and 7) exhibited remarkably stronger cytotoxic activity (IC50 values 0.39 μM and 0.32 μM, respectively) on keratinocytes than ingenol mebutate (IC50 value 0.84 μM). These compounds could serve as starting materials for further investigations to find alternatives to Picato® (with active substance ingenol mebutate), which was withdrawn from marketing authorization in the European Union.

Specialty Dihydrobenzoxanthone’s Artocarpus Purified By Vacuum Liquid Chromatography (VLC)

One family of plants that are source of bioactive chemicals is the Moraceae.Artocarpus is the main genus of the Moraceae. Several species of the genus Artocarpus have been isolated their secondary metabolites. The main fractions obtained from the VLC are analyzed again by TLC. Fractions that have same spots (Rf) pooled. Purification process on main factions are done repeatedly by radial chromatography. Flavonoid is the most found from Artocarpus plant. Dihydrobenzoxanthone is one of flavonoid derivatives which is successfully isolated from Artocarpus. Dihydrobenzoxanthone is only formed from the flavone with ring B which isoxygenated with pattern of 2', 4' and 5'. Students can be learned dihydrobenzoxanthone’s Artocarpus by laboratory activities.ÂÂ

3,6-dimethyl ester-α-mangostin Compound Modified from Isolate α-mangostin Garcinia Mangostana Linn

Isolate α-mangostin (1) is a phenolic compound derived from oxygenated and prenylated xanthones and major compounds obtained from the fruit peel of G. mangostana Linn., Which is stated to have inhibitory activity against α-glucosidase enzymes, serves to determine the antidiabetic activity resulting in IC50 value of 29,92 µM, is of the nature moderate to positive control (acarbose) with an IC50 value of 4.55 µM. A modified compound of α-mangostin (1) with acetic anhydride obtained by 3,6-di-methyl ester-α-mangostin (2) derivative showed the inhibitory value of α-glucosidase (IC50 13,89 μM), this value is better than the activity inhibition of α-mangostin (1), but not as active as the positive control value of the acarbose compound. The separation process to obtain α-mangostin isolates from the fruit peel of G. mangostana Linn was obtained by maceration method with ethyl acetate solvent, followed by refraction using a vacuum liquid chromatography (KCV) method over silica gel (Merck 60 G) and eluted using eluent (n-hexane: ethyl acetate) with increasing polarity, to produce as much pure crystal (21.66 g), yield (24%). While the structural characterization of the two compounds was carried out using UV-Vis, IR, HRESIMS, 1H-NMR and 13C-NMR spectroscopic methods, the antidiabetic testing was carried out using the α-glucosidase enzyme inhibition method.

Pharmacological activity of compounds isolated from methanolic extract marine sponge xestospongia sp. against escherichia coli and staphylococcus aureus

Few previous studies show pharmacological potencies of Xestospongia sp. and have not been explored further about its antibacterial activity. Thus, this study aims to isolate and identify the isolates from Xestospongia sp. and testing their antibacterial activity. Xestospongia Sp. was macerated with methanol then isolated and was purified by using vacuum liquid chromatography (VLC), radial chromatography (RC) and thin-layer chromatography (TLC). Isolated compounds then analyzed, identified, and determined their structures using 1H-NMR Spectrophotometer and by comparising data to references and ChemDraw 8.0. Compounds obtained with various concentrations (1000; 500; 100 μg/mL) then tested against Staphylococcus aureus ATCC 25923 and Escherichia coli ATCC 35218 using agar well-diffuse method. According to the study, two compounds isolated which were Saringosterol (isolate X1) and predicted-compound halenaquinone (isolate X2). The antibacterial test showed that isolate X1 was unable to inhibit bacteria‘s’ growth at each concentration, and isolate X2 showed antibacterial activity against S. aureus ATCC 25923 and E. coli ATCC 35218 at concentration 1000 and 500 μg/mL. In conclusion, 2 compounds successfully isolated from methanolic extract of Xestospongia sp. which were Saringosterol and predicted-compound halenaquninone, and only predicted-compound halenaquinone showed antibacterial activity at concentration 1000 and 500 μg/mL.

Antimicrobial activity of extracts and fractions of Euphorbia lateriflora (Schum. and Thonn) on microbial isolates of the urinary tract.

Euphorbia lateriflora is used in ethnomedicine for treating several conditions, including genital and urinary tract infections (UTI). Although ethnobotanical claims support its use in therapy, there is limited evidence on its effect on UTI, even though UTI remains a public health problem in Nigeria especially due to increasing antimicrobial resistance. We investigated the activity of E. lateriflora extracts and fractions on bacterial and fungal isolates from symptomatic urinary tract infections and vaginosis respectively. Qualitative phytochemical screening was conducted on dried pulverised leaves. Successive gradient extraction was carried out with the aid of a soxhlet extractor with n-Hexane, ethyl acetate and methanol respectively. Bioactivity guided fractionation was conducted on the ethyl acetate extract using Vacuum Liquid Chromatography. Antimicrobial susceptibility testing by disc diffusion was conducted on test isolates. Antimicrobial susceptibility of isolates to extracts and fractions was done using the agar well diffusion technique. Minimum Inhibitory Concentrations (MIC) and Minimum Biocidal Concentrations (MBC) were determined by agar and broth dilutions respectively. Time-kill assay of the ethyl acetate extract was conducted using the viable count technique. Phytochemicals present include saponins, tannins and flavonoids. The majority of isolates used in this study were multidrug resistant. Extracts and fractions of E. lateriflora produced appreciable zones of inhibition on both antibiotic susceptible and resistant bacteria with MICs of 6.25 mg/mL and MBC ranging from 6.25–50 mg/mL. Bactericidal activity of the ethyl acetate extract was concentration and time dependent with 100% kill at 25 mg/mL after 6 h for E. coli and 2 h for C. albicans. Euphorbia lateriflora contains phytochemicals which possess antimicrobial activity on antibiotic resistant bacteria and has potential in the development of chemotherapeutics for bacterial and fungal infections.

Stigmasterol and Stigmasterone from Methanol Extract of Calophyllum soulattri Burm. F. Stem Bark

Stigmasterol and Stigmasterone from Methanol Extract of Calophyllum soulattri Burm. F. Stem Bark. Calophyllum soulattri Burm. F. has been widely used for herbal medicine. Phytochemical investigation of C. soulattri contains a secondary metabolite of the steroid class. Steroid compounds have various biological activities, such as anti-inflammatory, antioxidant, antiproliferative, antibacterial, antimalarial, and anticancer. Two secondary metabolites steroids have been isolated and identified from the stem bark extract of C. soulattri. Isolation was carried out through the extraction (maceration), fractionation, and purification stages. Maceration is carried out using methanol as a solvent. Fractionation was carried out by vacuum liquid chromatography (VLC), and purification was by flash column chromatography. Identification of combined fractions and determination of pure isolates were used through thin-layer chromatography (TLC). The solvent used in the chromatography methods was a mixture of n-hexane and ethyl acetate. The structure isolates were identified by FTIR, 1H NMR, and 13C NMR and compared with literature data. Secondary metabolites steroids that have been isolated are identical compounds to stigmasterol and stigmasterone.

Potential Antibacterial Activity of Bioactive β-sitosterol from Root Bark of Rhizophora apiculata from Lampung Coastal

β-sitosterol is an essential bioactive phytosterol naturally present in plant cell membranes. It has a coincident structure with animal cholesterol. This investigation reported isolation, structure analysis, and an antimicrobial assay of β-sitosterol from the root bark of Bakau Minyak (Rhizophora apiculata) from Lampung coastal. The isolation of β-sitosterol was carried out through maceration using methanol, separation by vacuum liquid chromatography (VLC), and purification by column chromatography (CC) using ethyl acetate/n-hexane (2:8) as eluent. The structure of β-sitosterol was determined using spectroscopic analysis (UV-Vis, FT-IR, 13C-NMR, 1H-NMR, DEPT, and GC-MS). The pure β-sitosterol has 107.4 mg of white needle crystalline compound, the compound melting point about 140.7-141.2oC, the molecular mass confirmed by m/z 414, and UV absorption detected at λ 203.9 nm. The β-sitosterol antimicrobial bioactivity assay has shown potential activity to be developed as a lead compound against E. coli.

KARAKTERISASI STRUKTUR LIPID DARI SPONS XESTOSPONGIA TESTUDINARIA

The aim of this study is to isolate and characterize the structure of metabolite compounds of the sponge Xestospongia testudinaria, which grows in West Kalimantan. 3000 grams of sponges were analyzed through a preparation and extraction process, which produced a methanol-dissolved extract with a mass of 77.2 grams. The methanol-dissolved extract of was parted using n-hexane and dichloromethane as solvents. Based on the results of phytochemical testing, it can be concluded that the dissolved fraction of dichloromethane contains alkaloids and terpenoids or steroids. The purification process was carried out on 2.4 grams of dichloromethane-dissolved fraction by means of vacuum liquid chromatography, and ten separation fractions were obtained with eluated ethyl acetate : n-hexane as eluent. The separation fractions of FD 3, FD 4, FD 5 and FD 6, which have the same chromatogram characteristics were combined to be purified by means of preparative chromatography. The preparative process was carried out with chloroform : methanol 98:2 eluent, and three chromatogram isolates were produced. The chromatogram isolates with Rf = 0.44 and a mass of 8 milligrams were analyzed by FTIR spectroscopy using pellet KBr and 1H-NMR spectroscopy using CD3OD solvent. Based on the results of data analysis, it is concluded that the predicted content of metabolite compound of the Xestospongia testudinaria sponge sample growing in West Kalimantan waters is propyl-6,7,8-trihydroxy-4-pentadecenoate compound, which is a compound of the fat or lipid ester group with a typical IR absorption peak of the C = O group and C – O at wave numbers 1735.93 – 1766.80 cm-1 and 1111.00 – 1315.45 cm-1 and the containing 11 environments of hydrogen atoms scattered in the 1H-NMR chemical shift from 0.85 ppm to 3.97 ppm.

Identifikasi Senyawa Marker Dominan Ekstrak Daun Sirih Hitam dan Kuantifikasi Berdasarkan Perbedaan Lokasi Tanam

It has been revealed that the benefits of black betel leaf extract have created an opportunity for the black betel plant to be commercialized. The several benefits of black betel leaf extract are antimicrobial, anti-inflammatory, cytotoxic, and antioxidant. The extraction method uses maceration techniques. Identification of active markers as antimicrobials (Streptococcus mutans, Streptococcus sanguinis, Candida albicans) begins with Bioautography Thin Layer Chromatography (TLC), for isolation from macerated extracts followed by Vacuum Liquid Chromatography Fractionation (KCV), conventional Column Subfraction (KK), Layer Chromatography Thin Preparative (TLC). Quantification of marker compounds using a Thin Layer Chromatography (TLC) Scanner. Samples of black betel leaf (P. betle L. var Nigra) were collected from Samarinda (East Kalimantan), Wonosobo (Central Java), Godean (Yogyakarta), Banyuwangi (East Java) in June 2020. The results of the TLC test for bioautography of 70% ethanol extract and the ethanol fraction of 96% black betel leaf showed that the active spot of ethanol extract 70% for antibacterial activity on Streptococcus mutans Atcc 25175 and Streptococcus sanguinis Atcc 10556 was Rf 0.729 cm, the active spot of the ethanol fraction 96% for the activity of the fungus Candida albicans ATTC 10231; Rf 0.486 cm. Purification using TLC multi eluent with eluent I Hexane: Ethyl Acetate (7: 3) as non polar eluent and eluent II Chloroform: Ethyl Acetate (4: 1) as polar eluent on different TLC plates. The results obtained in the form of a single stain with Rf = 0.4 in eluent I and Rf = 0.73 in eluent II. The results of measuring the levels of the dominant marker compound using a TLC scanner showed that the highest marker compound area was from betel leaf grown in the Godean area (Yogyakarta) with the number 29869.4.

Antibacterial Potential by Rupture Membrane and Antioxidant Capacity of Purified Phenolic Fractions of Persea americana Leaf Extract.

The present research focused on evaluating the antibacterial effect and the mechanism of action of partially purified fractions of an extract of Persea americana. Furthermore, both its antioxidant capacity and composition were evaluated. The extract was fractionated by vacuum liquid chromatography. The antimicrobial effect against Staphylococcus aureus (ATCC 6538), Escherichia coli (ATCC 11229), Pseudomonas aeruginosa (ATCC 15442), and Salmonella choleraesuis (ATCC 1070) was analyzed by microdilution and the mechanism of action by the Sytox green method. The antioxidant capacity was determined by DPPH, FRAP, and ABTS techniques and the composition by Rp-HPLC-MS. All fractions showed a concentration-dependent antibacterial effect. Fractions F3, F4, and F5 (1000 µg/mL) showed a better antibacterial effect than the extract against the bacteria mentioned. The F3 fraction showed inhibition of 95.43 ± 3.04% on S. aureus, F4 showed 93.30 ± 0.52% on E. coli, and F5 showed 88.63 ± 1.15% on S. choleraesuis and 86.46 ± 3.20% on P. aeruginosa. The most susceptible strain to the treatment with the extract was S. aureus. Therefore, in this strain, the bacterial membrane damage induced by the extract and fractions was evidenced by light fluorescence microscopy. Furthermore, the extract had better antioxidant action than each fraction. Finally, sinensitin was detected in F3 and cinnamoyl glucose, caffeoyl tartaric acid, and cyanidin 3-O-(6′′-malonyl-3′′-glucosyl-glucoside) were detected in F4; esculin and kaempferide, detected in F5, could be associated with the antibacterial and antioxidant effect.

A bioactive compound isolated from Duku ( Lansium domesticum Corr) fruit peels exhibits cytotoxicity against T47D cell line.

Background: Breast cancer is a major health problem for women globally. Many attempts have been promoted to cure cancer by finding new anticancer medicines from natural resources. Despite the richness of biodiversity discovered, there are some natural resources that remain unexplored. Fruit peels of Duku ( Lansium domesticum Corr.) are rich with compounds that may have the potential to be developed as anticancer drugs. This study aimed to isolate cytotoxic compounds from the fruit peels of L. domesticum and assess their cytotoxic nature against T47D cells. Methods: Powdered peels were macerated with ethyl acetate and the filtrate was evaporated to give EtOAc extract A. Dried extract A was triturated with n-hexane to give n-hexane soluble fraction B and insoluble fraction C. The cytotoxic nature of these three samples were assessed using MTT assay using T47D cells and doxorubicin as a control. Results: Fraction C that showed the smallest IC50 (25.56 ± 0.64μg/mL) value compared to extract A and fraction B. Fraction C was further fractionated by vacuum liquid chromatography to give 6 subfractions. Subfraction 2 showed a single compound based on thin layer chromatography, and this compound was identified as Lamesticumin A on the basis of its spectroscopic data. Lamesticumin A demonstrated cytotoxic activity against T47D cell lines with an IC 50 value of 15.68± 0.30µg/mL. Conclusions: Further research is needed to investigate the potential of the natural compound Lamesticumin A derived from L. domesticum fruit peel as an anticancer therapy.