Isolation Of Secondary Metabolites Research Articles

Isolation of secondary metabolites from the mediterranean sponge species; Hemimycale columella and its biological properties

Despite the richness and biodiversity of invertebrates and algae in the Mediterranean Sea, these organisms are still poorly studied. The objective of our research is the discovery of bioactive lead compounds from the Mediterranean Sea sponge Hemimycale Collumella (HC). HC sponge (189.0 g) was collected from Mdiq costs on the Mediterranean Sea and extracted with methanol to yield (10 g) which was then subjected to fractionation. A bio-guided protocol was applied through evaluation of 2,2-diphenyl-1-picrylhydrazyl (DPPH) scavenging and Oxygen Radical Absorbance Capacity (ORAC), α-amylase, β-glucosidase, pancreatic lipase inhibition as well as anti-collagenase, anti-elastase, antityrosinase and cytotoxic activity. 2,3-O-Hexahydroxydiphenoyl-(α/β)-glucose (1) and gentisic acid 2-O-β-glucoside (2) were isolated from the water fraction, quercetin-3-O-β-glucopyranoside (3), kaempferol 3-O-β-glucopyranoside (4) and isorhamnetin 3-O-β-glucopyranoside (5) from n-butanol fraction, gallic acid (6) from ethyl acetate fraction and gallic acid-3-methyl ether (7) from methylene chloride fraction. Compound 5 had the highest DPPH and ORAC activity. Compounds 1–5 had promising lipase inhibition activities which exceeded that of the standard Orlistat, while compounds 1–7 showed anti-tyrosinase activity higher than that of the standard Hydroquinone monomethyl ether. This is the first report for evaluation of the biological activities of 2, 3-O-hexahydroxydiphenoyl-(α/β)-glucose (1), gentisic acid 2-O-β-glucoside (2) and gallic acid-3-methyl ether (7).

Phytochemical investigation of Pimpinella serbica

The plant species Pimpinella serbica (Vis.) Drude, endemic to West Balkans, belongs to the genus Pimpinella L. (Apiaceae) according to newer botanical classification. Initially, the plant was described as Pancicia serbica Vis. and long considered as a monotypic genus. Previous phytochemical investigations of this plant were limited to the essential oil analysis by the GC?MS method. This is first study that includes LC-DAD?MS screening of the extracts of the roots and aerial parts, followed by isolation of secondary metabolites using chromatographic techniques. Five compounds belonging to phenylpropanoids and polyacetylenes, well known for their bioactivities, were isolated and structurally determined by combined spectroscopic methods (UV, NMR, MS). Dillapiole (1), nothoapiole (2) and oplopantriol A 18-acetate (4) were found in both extracts, while falcarindiol (3) was isolated from the roots and dendrotrifidol (5) from the aerial parts only. The phytochemical profile of P. serbica L. supports its position in the Pimpinella L. genus.

Screening of in vitro antitrematodal activities of compounds and secondary metabolites isolated from selected Pteridophytes.

Four ferns Blechnum orientale Linn. (BO), Dicranopteris linearis (Burm.f ) Underw. (DL), Marattia fraxinea Sm. (MF), and Microlepia speluncae (L.) Moore (MS) were extracted in varied combination of organic solvents followed by the preparation of eluates and isolation of secondary metabolites using chromatography on a glass column with silica gel as the fixed phase. The chemical components were identified using HPTLC and GC-MS analysis. The in vitro anti-trematodal activities of these eluates and compounds were evaluated against the sheep trematode worm Gastrothylax crumenifer (Plagiorrchiida: Gastrothylacidae) at increasing concentrations (1 to 5 mg/mL), to find the relative motility (RM) values for 0 to 60 min of incubation as reflection of paralysis and death of the worms. Hedon-Fleig salt solution was used as negative control and Oxyclozanide® 1% as standard control. In vitro incubation study showed DL and MS extracts had strong trematodicidal activity. BO extract (5 mg/mL) produced moderate trematodicidal activity and MF (5 mg/mL) showed the least trematodicidal activity. Phytochemicals analysis revealed that the ferns are a potential source of trematodicidal compounds such as phytol isomers fern-8-ene and fern-9(11)-ene (terpenoid derivative), quercetin 7,3 ́,4 ́-trimethoxy (flavonoid derivative), etc., which offer scope for a more elaborate study for exploitation of ferns for human welfare.

Bioassay-Directed Isolation of Antibacterial Metabolites from an Arthropod-Derived Penicillium chrysogenum.

Bioassay-directed isolation of secondary metabolites from an extract of Penicillium chrysogenum TJ403-CA4 isolated from the medicinally valuable arthropod Cryptotympana atrata afforded five new and 10 known compounds (1-15). All the compounds (except 14) belong to a minor class of highly rigid 6-5-5-5-fused tetracyclic cyclopiane-type diterpenes known to be exclusively produced by members of the Penicillium genus. The structures and absolute configurations of the new compounds (1-5) were elucidated by extensive spectroscopic analyses, including HRESIMS and 1D and 2D NMR, single-crystal X-ray diffraction, and comparison of the experimental electronic circular dichroism data. Compounds 1 and 2 represent the first examples of cyclopianes bearing a C-20 carboxyl group; compound 3 represents the first example of a cyclopiane with a gem-hydroxymethyl group; compound 4 represents the second example of a cyclopiane bearing a hydroxy group at C-7; compound 5 represents the first example of a cyclopiane bearing a hydroxy group at C-8. Compounds 2 and 3 exhibited activity against MRSA, with MIC values of 4.0 and 2.0 μg/mL, respectively. In addition, the structure-antibacterial activity relationship (SAR) of compounds 1-15 is discussed.

The isolation of secondary metabolites from Rheum ribes L. and the synthesis of new semi-synthetic anthraquinones: Isolation, synthesis and biological activity

Rheum ribes L. (Rhubarb) is one of the most important edible medicinal plants in the Eastern Anatolia region and is called “Işkın” by local people. Resveratrol and 6-O-methylalaternin were isolated from the Rhubarb for the first time in addition to well-known secondary metabolites including emodin, aloe-emodin, β-sitosterol and rutin. The new semi-synthetic anthraquinone derivatives with the NαFmoc-l-Lys and ethynyl group were synthesized from the isolated anthraquinones emodin and aloe-emodin of Rhubarb to increase the bioactivities. Aloe-emodin derivative with NαFmoc-l-Lys shows the highest inhibition values by 94.11 ± 0.12 and 82.38 ± 0.00% against HT-29 and HeLa cell lines, respectively, at 25 µg/mL. Further, modification of the aloe-emodin with both the ethynyl and the NαFmoc-l-Lys groups showed an antioxidant activity-enhancing effect. From molecular docking studies, the relative binding energies of the emodin and aloe-emodin derivatives to human serum albumin ranged from −7.30 and −10.62 kcal/mol.

Analysis and Isolation of Secondary Metabolites of Bipolarissorokiniana by Different Chromatography Techniques and the Spectrum of Their Biological Activity

Secondary metabolites were isolated from cultures of four geographically distant isolates of the cosmopolitan fungus, Bipolaris sorokiniana for intense biological characterization. Analysis of extracts from their liquid and solid cultures with TLC and HPLC-MS showed high similarity of their composition despite on fungal origin. From the extracts of B. sorokiniana 14 metabolites (terpenes, quinones, xanthones etc.) were isolated using column chromatography and preparative HPLC; among them, 12 known substances were identified. Chloromonilinic acid B (CAB) showed the highest level phytotoxic activity on wheat leaves. The most of isolated compounds inhibited growth of Bacillus subtilis, of which methyl-8,3-hydroxy-3-methyl-4-chloroxanthon-1-carboxilate (MMC) demonstrated the strongest activity. Some metabolites of B. sorokiniana (CAB and MMC) were shown to have insecticidal activity against common cereal aphid. All the compounds were cytotoxic (especially, cochlioqionones and fusaroproliferin) when tested on Sf9 cell line. However, no one was acute toxic for infusoria. Evidently, the production of multiple bioactive compounds makes B. sorokiniana a strong competitor in different ecological niches and a dangerous contaminant of grain.

In vitro propagation and assessment of genetic fidelity in Dioscorea deltoidea, a potent diosgenin yielding endangered plant

Dioscorea deltoidea (Family: Dioscoreaceae) is a high value endangered medicinal plant and the source of pharmaceutically important bioactive compound diosgenin, which is used in the synthesis of steroidal drugs. Establishing an efficient reproducible regeneration system is necessary for its sustainable utilization and conservation. The purpose of this research was to establish an effective and efficient in vitro regeneration protocol for D. deltoidea and assess its genetic and biochemical fidelity. In this report, Murashige and Skoog (MS) medium fortified with different types and concentration of Plant Growth Regulators (PGRs) were studied. Maximum shoot regeneration frequency was achieved in MS medium supplemented with 2.0 mg L − 1 BA whereas higher shoot multiplication was observed in 2.0 mg L − 1 BA + 1.0 mg L − 1 IBA. Highest rooting was recovered on MS medium supplemented with 2.0 mg L − 1 NAA. In vitro rooted plantlets were acclimatized with a survival rate of 96%. The genetic fidelity of the mother plant (MP), acclimatized plant (AP) and in vitro regenerants were found to be clonally similar as examined through ISSR fingerprinting. Of 7 ISSR primers tested, 5 produced scorable amplified products. The High Performance Thin Layer Chromatography (HPTLC) based chemical assessment showed varied chemical profile. The developed protocol would be beneficial in providing a germplasm conservation system, isolation of secondary metabolites and production of high value clonal regenerants for commercial production.

Phytochemical and Biological Investigation of Curcuma amada Leaves

Methanolic extract of Curcuma amada leaves was investigated for isolation of secondary metabolites by successive chromatographic separation (VLC, column chromatography and TLC) which yielded several purified compounds. Among them the structures of β-sitosterol and stigmasterol were determined by analysis of high resolution 1H NMR spectroscopic data and co-TLC with authentic samples. The crude methanolic extract of C. amada leaves and its different fractionates i.e. petroleum ether (PESF), chloroform (CSF) and aqueous (AQSF) soluble fractions were evaluated for antibacterial, antioxidant, cytotoxic and thrombolytic activities. In antibacterial screening, the CSF exhibited the highest inhibition against bacterial growth having zone of inhibition 12 mm compared to the standard kanamycin where the zone of inhibition was observed at 18 mm against gram negative Vibrio mimicus. Significant free radical scavenging activity was also exhibited by CSF with the IC50 value of 103.09 μg/ml as compared to tert-butyl-1-hydroxytoluene (BHT) having IC50 31.88 μg/ml. In cytotoxicity study, the crude extract showed significant lethality towards brine shrimp having with the LC50 value of 6.540 μg/ml as compared to the standard vincristine sulfate (0.451μg/ml). In the study for thrombolytic property, different extracts of C. amada exhibited clot lysis ranging from 17.24 to 43.55% as compared to standard streptokinase (93.75 %).
 Dhaka Univ. J. Pharm. Sci. 19(1): 9-13, 2020 (June)

Potential of peel extracts of Punica granatum and Citrus aurantifolia on alloxan-induced diabetic rats

BackgroundPeel is one of the major by-products in fruit processing industry. Fruit products (non-edible parts) are also considered as waste products and often discarded in the environment. Fruit peels are now serving as one of the primary sources for isolation and extraction of secondary metabolites in pharmaceutical industry. The present investigation was carried out to screen the phytochemical constituents and HPTLC analysis of peel extracts of Punica granatum and Citrus aurantifolia and their antidiabetic potential in alloxan-induced diabetic rats.ResultsAmong the different solvent extracts, methanol solvent extract was found to possess more amounts of secondary metabolites. In addition, HPTLC analysis of the plant samples revealed the presence of 13 peaks in both the plants by using gallic acid as marker. Different biochemical parameters such as blood glucose, cholesterol, protein, urea, creatinine, and triglycerides level were subjected for estimation by collecting the blood samples from the treated diabetic rats after 21 days. A sharp decline in blood glucose, cholesterol, triglycerides, creatinine, and urea level was noticed when methanolic extracts of Punica granatum and Citrus aurantifolia were given to experimental animals when compared with negative control. However, protein and weight of the animal were found to be enhanced when treated with methanolic extracts of both the plants.ConclusionIt can be concluded that fruit peels of both the plants exhibited antidiabetic potential on alloxan-induced diabetic rats which can be attributed to wide range of active pool of secondary metabolites. Further, screening and isolation of secondary metabolites along with their mode of action is required for effective use of plant-based drugs as antihyperglycemic agent.Graphical abstract

PHTYOCHEMICAL SCREENING AND BIOCHEMICAL ANALYSIS OF SELECTED HERBAL PLANTS

Documentation of the herbal plants in the Department of Education’s Herbal garden serves as an information dissemination for teachers and students and an avenue to actually know more of the herbal plants. This study aimed to document the herbal plants grown in the herbal gardens of the elementary schools in the Department of Education. At the same time, determined the herbal plants grown which through literature review showed lack of study. Through this, it will ascertain the phytochemical and biochemical analysis of the said selected herbal plants for its safety and efficacy. The study revealed that Mussaenda philippica var. Alicia, Setaria palmitolia, Justicia gendarussa Burm. f., Ageratum conyzoides L., and Mussaenda anisophylla s. Vidal contain secondary metabolites present in the leaf extracts of these herbal plants which include alkaloids, leucoanthocyanin as flavonoid compound, glycosides, saponins and tannins. The ethanol leaf extracts possess superior microbial activity and high antioxidant capacity. Thus, this study presented the therapeutic potentials of the herbal plants for further research on extracts for identification, purification and isolation of secondary metabolites having antimicrobial properties and antioxidant activity.

Plant Extracts and their Secondary Metabolites as Modulators of Kinases.

Natural Products (NP), specifically from medicinal plants or herbs, have been extensively utilized to analyze the fundamental mechanisms of ultimate natural sciences as well as therapeutics. Isolation of secondary metabolites from these sources and their respective biological properties, along with their lower toxicities and cost-effectiveness, make them a significant research focus for drug discovery. In recent times, there has been a considerable focus on isolating new chemical entities from natural flora to meet the immense demand for kinase modulators, and also to overcome major unmet medical challenges in relation to signal transduction pathways. The signal transduction systems are amongst the foremost pathways involved in the maintenance of life and protein kinases play an imperative part in these signaling pathways. It is important to find a kinase inhibitor, as it can be used not only to study cell biology but can also be used as a drug candidate for cancer and metabolic disorders. A number of plant extracts and their isolated secondary metabolites such as flavonoids, phenolics, terpenoids, and alkaloids have exhibited activities against various kinases. In the current review, we have presented a brief overview of some important classes of plant secondary metabolites as kinase modulators. Moreover, a number of phytocompounds with kinase inhibition potential, isolated from different plant species, are also discussed.

Isolation of Secondary Metabolites from Leucas aspera and Investigation of Biological Activity

Leucas aspera plant was subjected to isolation of secondary metabolites and screening of their biological activities. Four compounds, stigmasterol, lupeol, β-sitosterol and menthol, were isolated from methanol extract. Sixteen different micro-organisms were used for investigating antimicrobial activity of the different extracts of L. aspera where noteworthy zone of inhibition was observed against Gram positive B. subtilis and S. aureus, B. megaterium and Gram negative S. paratyphi, S. typhi, V. mimicus, S. dysenteriae and V. cholera. In brine shrimp lethality bioassay, the highest lethality was showed by crude methanol extract having LC50 values of 4.07μg/mL. The total antioxidant capacity of crude methanol fraction was found to be 59.40 mg/g of plant extract which was maximum comparing with other fractions. No significant cytotoxicity was observed on both HeLa and Vero cell at 1mg/mL sample inhibition.
 Dhaka Univ. J. Sci. 68(1): 101-104, 2020 (January)

Natural Product Chemistry Laboratory: Isolation Phenobarbitone from Mangrove Bark (<i>Avicennia marina</i>)

Natural product chemistry laboratory in this research carried out a project-based laboratory related to the isolation of secondary metabolites from medicinal plants. The students learn the essential skills required to perform the extraction, fractionation, purification, and structural elucidation of secondary metabolites from medicinal plant. This study was conducted to isolate phenobarbitone from the bark of A. marina. The modified method uses maceration and recrystallization. This study obtained phenobarbitone 0.102 g from 100 g dried powder of bark of A. marina. Identified of phenobarbitone based on chromatogram of TLC, FT-IR, and UV-Vis.

ANTIOXIDANT ACTIVITY of THE SECONDARY METABOLITES PRODUCED by ENDOPHYTIC FUNGI ISOLATED from JERUJU (Acanthus ilicifolius L.) PLANT

Twelve endophytic fungi were isolated from Jeruju plant. Three of which have the best antioxidant activity, namely Culvularia lunata DJS1, Diaporthe phaseolorum DJS2, and Colletotrichum siamense BJS4. The fungi can produce secondary metabolites that have the potential as antioxidant compounds. This study aims to isolate the antioxidant compound using a chromatography column and test the antioxidant activity using DPPH method. Isolation of secondary metabolites produced 40 eluates. The IC50 value of DJS111 and BJS42 eluates were 10.20 µg/ml and 20.89 µg/ml, which is an antioxidant compound with very strong activity. The IC50 value of DJS24 eluate was 54.45 µg/ml, which is strong antioxidant activity. DJS111 eluate has antioxidant activity that approaches to ascorbic acid as a standart antioxidant compound (7.76µg/ml). From TLC, DJS111 contains flavonoid compound and DJS24 and BJS42 contain terpenoid compound.

Antibacterial Activity of The Secondary Metabolites Produced by Endophytic Fungi Isolated from Acanthus ilicifolius Plant

Twelve endophytic fungi were isolated from the Acanthus ilicifolius plant. Two of them have the best antibacterial activity, namely Culvularia lunata and Diaporthe phaseolorum. Fungi can produce secondary metabolites that have the potential as antibacterial compounds. This study aims to isolate antibacterial compounds using column chromatography methods and test the antibacterial activity using diffusion methods to use paper discs. Isolation of secondary metabolites produces 27 eluates. From TLC, it is known that the endophytic fungi Culvularia lunata produce an active compound as an antibacterial that is phenol compound. Endophytic fungi Diaporthe phaseolorum produces three types of active compounds as antibacterial namely phenol, terpenoids, and alkaloids, each component of the active compound are tested for sensitivity to bacterias Escherichia coli, Staphylococcus aureus, Shigella dysenteriae, and Salmonella typhi. MIC values ??of terpenoid compounds with concentrations of 125-250 ppm are antibacterial compounds with good activity against Escherichia coli, Staphylococcus aureus, Shigella dysenteriae, and Salmonella typhi.

Terpenoids and phytosteroids isolated from Commelina benghalensis Linn. with antioxidant activity.

Background Commelina benghalensis Linn. (Family: Commelinaceae) is a common weed available in Bangladesh with several uses in traditional medicine. However, the chemical profile of this medicinal plant is scarce in relation to its medicinal uses. The aerial parts of this plant have been investigated for the isolation of secondary metabolites and evaluation of the biological activities. Methods Major phytochemical groups were analyzed using chromogenic reagents, whereas n-hexane soluble fractionates of the methanol extract were subjected to 1H nuclear magnetic resonance (NMR) spectroscopic analysis. The antioxidant property of the obtained compounds was evaluated using 1,1-diphenyl-2-picryl-hydrazyl (DPPH). Results Dammara-12-en-3-one (CB-1), stigmasterol (CB-2) and 3 (2,3,4,5,6-pentahydroxy)-cinnamoyl dammara-12-ene (CB-3) were isolated from the n-hexane fractionate of methanol extract of C. benghalensis. In the study of DPPH radical scavenging activity, IC50 values were predicted to be 790.18, 4186.94 and 2001.16 μg/mL for CB-1, CB-2 and CB-3, respectively, whereas standard ascorbic acid showed IC50 at 1.26 μg/mL. Conclusions Two new dammarane-type triterpene (CB-1 and CB-3) and one phytosterol (CB-2) were identified in C. benghalensis with mild antioxidant property.

Isolation of secondary metabolites from Geranium molle L. with anticancer potential

Two flavone glycosides, apigenin-6-C-glucoside (1) and lutelin-6-C-glucoside (2), and an anomeric mixture of 2,3-dygalloyl-4,6-HHDP-d-glucose (3a/3b) were isolated for the first time from Geranium molle L., a medicinal plant used in the folk medicine of northeast of Portugal for the treatment of a variety of ailments, including cancer. The cytotoxic activity of the isolated compounds was assessed against several human tumour cell lines and non-tumour porcine liver primary cells. The anomeric mixture of ellagitannins 3a/3b, with the two anomers showing considerably different hydrophobicities, was found to be significantly cytotoxic against cancer cells, with only moderate hepatotoxicity.

Recent metabolomics and gene editing approaches for synthesis of microbial secondary metabolites for drug discovery and development.

Microbial secondary metabolites (SMs) have been identified as an important natural source of drugs for several metabolic and neurological diseases. Along with biomedical applications, SMs are also used in the food and biochemical industries. SMs include natural products such as pigments, alkaloids, toxins, antimicrobials obtained from cultured microorganisms, while other non-cultivable microorganisms have also acted as a rich source of SMs. But, the isolation of SMs from these sources is a very tedious task. Metabolomics provides complete identification and structural information about the entire cellular metabolome under specific conditions using highly sophisticated instrumentation. Further, gene editing techniques such as cloning and gene refactoring, including advanced CRISPR-Cas, can be used for engineering microbes that have the potential to produce natural SMs that were not produced in native microbial strain. The present review describes integrated metabolomics and gene editing approaches for the synthesis of novel microbial SMs and their potential application towards drug discovery and development.

The Phylum Bryozoa as a Promising Source of Anticancer Drugs.

Recent advances in sampling and novel techniques in drug synthesis and isolation have promoted the discovery of anticancer agents from marine organisms to combat this major threat to public health worldwide. Bryozoans, which are filter-feeding, aquatic invertebrates often characterized by a calcified skeleton, are an excellent source of pharmacologically interesting compounds including well-known chemical classes such as alkaloids and polyketides. This review covers the literature for secondary metabolites isolated from marine cheilostome and ctenostome bryozoans that have shown potential as cancer drugs. Moreover, we highlight examples such as bryostatins, the most known class of marine-derived compounds from this animal phylum, which are advancing through anticancer clinical trials due to their low toxicity and antineoplastic activity. The bryozoan antitumor compounds discovered until now show a wide range of chemical diversity and biological activities. Therefore, more research focusing on the isolation of secondary metabolites with potential anticancer properties from bryozoans and other overlooked taxa covering wider geographic areas is needed for an efficient bioprospecting of natural products.

Lantana camara: Secondary Metabolite Isolation by Analytical Techniques

The Plant Lantana camara belongs to the family Verbenaceae, have always been an important source of phytomedicinal agents since ancient times, Until today, it continue to provide modern medicine with novel treatments and to support to identify and isolate compounds from Indian flora with potential biological activity and medicinal value. It has been reported to be used in folk remedies For instance, used for antibacterial, antiulcer, antioxidant, and also treatment for malaria, rheumatisms, asthma, tumors. Many Literature review and phytochemical investigations have been done on this plant, reported to contain various compounds like triterpenoids, proteins, carbohydrates, lactones, furfural, flavonoids, alkaloids, glycosides, tannins, steroids. The ethanolic extract were subjected for column chromatography for the isolation of secondary metabolites by using stationary phase as silica gel with mesh number of 230-400 and the mobile phase was 20% & 30% ethyl acetate/hexane. The Functional groups, structural analysis of the isolated metabolites identified from IR spectrum resembled functional groups of flavonoid chemical structure, Yellow color is characteristic of flavonoids. Keywords: Lantana camara, secondary metabolites, Column chromatography, TLC, IR spectroscopy.