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Thin layer chromatography bioautographic assays facilitate the acquisition of activity-profile chromatograms and assist in pinpointing active constituents within complex mixtures by observing the inhibition halos they produce. Peroxidase is an enzyme implicated in the browning of different fresh cut vegetables and in several diseases. A peroxidase bioautographic assay was developed, based on enzyme agarose immobilization and the 2,2′-Azino-bis(3-ethylbenzothiazoline-6-sulfonic acid) diammonium salt/radical cation (ABTS/ABTS·+) reporter system. Peroxidase was purified from potatoes with the aim to detect specific inhibitors. To reduce false positives, a non-enzymatic assay was also employed. The best results are obtained when a solution containing agarose, ABTS, hydrogen peroxide, and peroxidase in phosphate buffer is poured over the TLC plate (final concentrations: 0.031 mmoles/cm2, 0.239 µmoles/cm2, and 84.04 U/cm2) and incubated for 70 min. Limit of detection and quantification for quercetin is 0.16 µg and 0.54 µg, respectively. The developed system is able to detect quercetin in a Solidago chilensis Meyen extract and a peroxidase inhibitor in a Cichorium intybus L. extract. Therefore, the assay can detect inhibitory constituents in complex mixtures and differentiate between peroxidase inhibitors and ABTS·+ radical scavengers before any preparative fractionation, helping to take early operational decisions that can save time and resources. [ABSTRACT FROM AUTHOR]

Background/Objective: Numerous studies have demonstrated the ethno-and pharmacological properties of various Artemisia species. However, this plant genus which grows abundantly and wildly in the 'Tébessa' Algerian region has not been investigated. This study focuses on the phytochemical characterization, biological activities evaluation with the antibacterial fraction identification of the aerial part aqueous extract (AE) and essential oil (EO) from Algerian Artemisia herba alba (Aha) growing wild in the 'Tebessa' region. Methods: Polyphenols, flavonoids, and condensed tannins were analyzed by spectrophotometer and the antimicrobial activity was determined by the gel diffusion followed by micro-dilution methods. The antioxidant effect was assayed using 2,2-diphenyl-1-picrilhidrazil (DPPH), total antioxidant capacity (TAC), and ferric reducing power (FRAP) tests. The cytotoxic and anti-leishmaniasis activities were experienced on Raw 264.7 cell line and on L. infantum and L. major promastigotes respectively. Thin-layer chromatography (TLC)/autobiography and gas chromatography-mass spectrometry (GC-MS) techniques were used to isolate and identify the antibacterial compounds. EO profile was determined by GC-MS analysis. Results: The extracts (AE and EO) demonstrated antioxidant activity, with the AE exhibiting the most pronounced levels. Both extracts showed antimicrobial activity against all the strains tested however; the EO effect was more effective, especially against the E. feacalis strain with a minimum inhibitory concentration (MIC) value of 0.0625 mg/mL. The antibacterial fraction exerted by AE towards E. feacalis was identified. AE and EO extracts showed antileishmanial activity and no toxic effect however, volatile extract was the most effective. EO profile revealed 19 identified compounds, led by camphor (51.14%), followed by 1,8 Cineole (19.30%), camphene (6.90%) and chrysanthenone (5.68%). Conclusion: As a result, the both extracts promise a high potential applications in agri-food, pharmaceutical, cosmetic sectors. This study aligns with Sustainable Development Goals (SDGs) 3, 12, and 15, emphasizing potential health benefits, the responsible use of natural resources, and biodiversity conservation. [ABSTRACT FROM AUTHOR]

Infectious diseases pose a formidable global challenge, compounded by the emergence of antimicrobial resistance. Consequently, researchers are actively exploring novel antimicrobial compounds as potential solutions. This endeavor underscores the pivotal role of methods employed for screening and evaluating antimicrobial activity—a critical step in discovery and characterization of antimicrobial agents. While traditional techniques such as well-diffusion, disk-diffusion, and broth-dilution are commonly utilized in antimicrobial assays, they may encounter limitations concerning reproducibility and speed. Additionally, a diverse array of antimicrobial assays including cross-streaking, poisoned-food, co-culture, time-kill kinetics, resazurin assay, bioautography, etc., are routinely employed in antimicrobial evaluations. Advanced techniques such as flow-cytometry, impedance analysis, and bioluminescent technique may offer rapid and sensitive results, providing deeper insights into the impact of antimicrobials on cellular integrity. However, their higher cost and limited accessibility in certain laboratory settings may present challenges. This article provides a comprehensive overview of assays designed to characterize antimicrobial activity, elucidating their underlying principles, protocols, advantages, and limitations. The primary objective is to enhance understanding of the methodologies designed for evaluating antimicrobial agents in our relentless battle against infectious diseases. By selecting the appropriate antimicrobial testing method, researchers can discern suitable conditions and streamline the identification of effective antimicrobial agents. [ABSTRACT FROM AUTHOR]

Poplars provide medicinal raw plant materials used in pharmacy. Leaf buds are one of the herbal medicinal products collected from poplars, having anti-inflammatory and antiseptic properties, but there are no quality standards for their production and there is a need to determine their botanical sources. Therefore, the chemical compositions of the leaf buds from four species and varieties of poplars, Populus balsamifera, P. × berolinensis, P. × canadensis 'Marilandica', and P. wilsonii were investigated and compared using gas chromatography coupled with mass detection (GC-MS) and two-dimensional high-performance thin-layer chromatography (2D-HPTLC) in order to search for taxa characterized by a high content of biologically active compounds and with a diverse chemical composition that determines their therapeutic effects. The presence of 163 compounds belonging to the groups of flavonoids, phenolic acids derivatives, glycerides, and sesquiterpenes was revealed. Moreover, the conditions for the separation and identification of biologically active compounds occurring in analyzed leaf buds using 2D-HPTLC were optimized and used for metabolomic profiling of the studied poplars, enabling their fast and simple botanical identification. The total phenolic (TPC) and flavonoid (TFC) contents of examined extracts were determined and their antioxidant capacities were estimated by spectrophotometric DPPH, ABTS, and FRAP assays. Based on the analysis of phytochemicals and antioxidant activity, P. × berolinensis buds were selected as the raw plant material for medicinal purposes with the highest content of active compounds and the strongest antioxidant activity. [ABSTRACT FROM AUTHOR]

Currently, the antimicrobial activity of essential oils (EOs) is an outstanding research field due to antibiotic resistance of microorganisms. Thin-layer chromatography‒direct bioautography (TLC‒DB) is an effective, fast method to find components with antimicrobial activity in a mixture of plant compounds, e.g., in EOs. The volatility and hydrophobic characters of EOs require special experimental conditions, and disc diffusion assay is not appropriate to explore the antimicrobial activity of them. The aim of this study was to use "R" mutants, which are more sensitive to synthetic antimicrobial drugs, in DB to increase the sensitivity of this method. Our hypothesis was that these mutants show sensitivity to some EOs (thyme, clove, and peppermint) as well. The chemical composition of our tested EOs was measured with gas chromatography‒mass spectrometry (GC‒MS). The main compounds (39.8% thymol, 78.8% eugenol, and 50.4% menthol) of EOs showed notable antibacterial activity in TLC‒DB. Based on our results, we suggest to use Salmonella minnesota Re595 rough strain as test bacterium in bioautography, because it showed the highest sensitivity to the tested antibiotics (gentamicin and cephalexin) and EOs. Furthermore, this rough mutant could make TLC‒DB more faster, because only 4 h incubation time was enough to detect the inhibition zones of the active compounds used in this study. [ABSTRACT FROM AUTHOR]

For non-target residue analysis of xenoestrogens in food, sophisticated chromatographic–mass spectrometric techniques lack in biological effect detection. Various in vitro assays providing sum values encounter problems when opposing signals are present in a complex sample. Due to physicochemical signal reduction, cytotoxic or antagonistic effect responses, the resulting sum value is falsified. Instead, the demonstrated non-target estrogenic screening with an integrated planar chromatographic separation differentiated opposing signals, detected and prioritized important estrogenic compounds, and directly assigned tentatively the responsible compounds. Sixty pesticides were investigated, ten of which showed estrogenic effects. Exemplarily, half-maximal effective concentrations and 17β-estradiol equivalents were determined. Estrogenic pesticide responses were confirmed in six tested plant protection products. In food, such as tomato, grape, and wine, several compounds with an estrogenic effect were detected. It showed that rinsing with water was not sufficient to remove selected residues and illustrated that, though not usually performed for tomatoes, peeling would be more appropriate. Though not in the focus, reaction or breakdown products that are estrogenic were detected, underlining the great potential of non-target planar chromatographic bioassay screening for food safety and food control. [ABSTRACT FROM AUTHOR]

The bulbs of Allium parvum are used as a flavoring agent in diet and are conventionally assumed to treat skin infections. The current study evaluates the phytochemicals from organic extract of A. parvum and their antifungal activity against biofilm-forming dermatophytic fungi Malassezia furfur MTCC 1374, yeast found in the normal flora of the skin, causes superficial infections, systemic infections like pulmonary infections, fungemia, etc., and implant-associated infection by biofilm formation. Biofilm formation enables the fungi to escape from antifungal agents and renders resistance to antifungal agents. The phytochemical assay for the organic extract of the onion bulb documented that A. parvum contains phenol (58 + 0.1 mg GAE/g), flavonoids (112 + 0.12 mg QE/g), saponins (1.45 + 0.1 mg AE/g), and tannins (4.1 + 0.03 g TA/g). The FTIR and GC-MS analysis revealed the presence of specific compounds, alkaloids such as glycosides and quinolones, plant flavonoids such as 4H-Pyran-4-one, 2,3-dihydro-3,5-dihydroxy-6-methyl-ester, phenolics, saponins such as butanoic acid and gluconic acid, and organic sulfur compounds such as diallyl sulfide, thiopyran tetrahydro-ester, allyl methyl trisulfide, and allyl methyl di sulfide. These compounds inhibited the growth of M. furfur at a minimum inhibitory concentration (MIC) value of 6.25 mg/ml and its biofilm at a minimum biofilm inhibitory concentration (MBIC) value of 12.5 mg/ml. Further studies required to explore the possible mechanism in controlling the biofilm, so, as to recommend as fungal control agent in medical sector. [ABSTRACT FROM AUTHOR]

Quality assessment is the most important step in the production of effective and safe food supplements. The aim of this study is to develop a green, simple, and high-throughput procedure for the quality assessment of Origanum vulgare L. commercial products using high-performance thin-layer chromatography (HPTLC)/bioautography with a multicriteria decision method, sum of ranking differences (SRD). Direct bioautography assays were developed for the identification of constituents with antibacterial activity tested against Gram-positive (Bacillus subtilis, Micrococcus lysodeikticus) and Gram-negative (Escherichia coli) strains, antioxidative (DPPH) activity, and enzyme (α-amylase) activity. Combination of the leave-one-out cross-validation methodology of SRD values and significant differences by post hoc Wilcoxon matched pairs test were used to evaluate commercial varieties. It was proved that suggested bioautography/chemometric screening methodology could be used as guidelines for the selection of any theoretical ideal product. Analysts may have benefits from this time-saving and streamlined quality profiling. [ABSTRACT FROM AUTHOR]

The bulbs of Allium parvum are used as a flavoring agent in diet and are conventionally assumed to treat skin infections. The current study evaluates the phytochemicals from organic extract of A. parvum and their antifungal activity against biofilm-forming dermatophytic fungi Malassezia furfur MTCC 1374, yeast found in the normal flora of the skin, causes superficial infections, systemic infections like pulmonary infections, fungemia, etc., and implant-associated infection by biofilm formation. Biofilm formation enables the fungi to escape from antifungal agents and renders resistance to antifungal agents. The phytochemical assay for the organic extract of the onion bulb documented that A. parvum contains phenol (58 + 0.1 mg GAE/g), flavonoids (112 + 0.12 mg QE/g), saponins (1.45 + 0.1 mg AE/g), and tannins (4.1 + 0.03 g TA/g). The FT-IR and GC-MS analysis revealed the presence of specific compounds, alkaloids such as glycosides and quinolones, plant flavonoids such as 4H-Pyran-4-one, 2,3-dihydro-3,5-dihydroxy-6-methyl-ester, phenolics, saponins such as butanoic acid and gluconic acid, and organic sulfur compounds such as diallyl sulfide, thiopyran tetrahydro-ester, allyl methyl trisulfide, and allyl methyl di sulfide. These compounds inhibited the growth of M. furfur at a minimum inhibitory concentration (MIC) value of 6.25 mg/ml and its biofilm at a minimum biofilm inhibitory concentration (MBIC) value of 12.5 mg/ml. Further studies required to explore the possible mechanism in controlling the biofilm, so, as to recommend as fungal control agent in medical sector.

Abstract Background Sphaeranthus indicus L. is a well-known medicinal plant in folk medicine. A variety of biological activities and chemical substances in this plant have been reported. The phytochemical content and activity may vary according to geographic location. This study aims to determine the geographical significance, the analysis of the synergistic effect of phytochemicals, the identification of active compounds, and the determination of the action mechanism of S. indicus inflorescence methanolic extract against Staphylococcus aureus and Klebsiella pneumonia. Results The bands with Rf values of 0.92 and 1.0 showed antimicrobial activity, while all bands showed antioxidant activity. The first fraction showed the highest antimicrobial activity, and the pool of the second fraction showed the highest antioxidant activity. The kinetics of the antioxidants differed among the fractions. Analysis of synergistic effects showed that several compounds were involved in the activities. The bands with Rf 0.45, 0.55, 0.68, 0.79, and 0.85 were active components of the extract. Leakage of cell contents was detected at 260 and 280 nm wavelengths. Six different proteins and one nucleic acid band were detected after electrophoresis. The SEM analysis showed that the phytochemicals caused severe membrane damage. Conclusion The study revealed that the photochemical present in methanol extract of the inflorescence of S. indicus has a synergistic effect and acts on bacterial cell envelope. The five compounds were identified as active molecules belonging to the class of terpenoids. The result also signified the geographical area since thymol was identified for the first time in this plant at this location. Graphical abstract

Natural products and their analogues have contributed significantly to treatment options, especially for anti-inflammatory and infectious diseases. Thus, the primary objective of this work was to compare the bioactivity profiles of selected medicinal plants that are historically used in folk medicine to treat inflammation and infections in the body. Chemical HPTLC fingerprinting was used to assess antioxidant, phenolic and flavonoid content, while bioassay-guided HPTLC was used to detect compounds with the highest antibacterial and anti-inflammatory activities. The results of this study showed that green tea leaf, walnut leaf, St. John's wort herb, wild thyme herb, European goldenrod herb, chamomile flower, and immortelle flower extracts were strong radical scavengers. Green tea and nettle extracts were the most active extracts against E. coli, while calendula flower extract showed significant potency against S. aureus. Furthermore, green tea, greater celandine, and fumitory extracts exhibited pronounced potential in suppressing COX-1 activity. The bioactive compounds from the green tea extract, as the most bioactive, were isolated by preparative thin-layer chromatography and characterized with their FTIR spectra. Although earlier studies have related green tea's anti-inflammatory properties to the presence of catechins, particularly epigallocatechin-3-gallate, the FTIR spectrum of the compound from the most intense bioactive zone showed the strongest anti-inflammatory activity can be attributed to amino acids and heterocyclic compounds. As expected, antibacterial activity in extracts was related to fatty acids and monoglycerides. [ABSTRACT FROM AUTHOR]

The leaves of Ficus benghalensis var. krishnae, a tree endemic to India, are known for their unique cup-shaped appearance and have been used in traditional medicine. The present study examines the antimicrobial activity of leaf extracts prepared in various solvents against four bacterial species, namely Bacillus subtilis, Escherichia coli, Salmonella typhimurium,, and Staphylococcus aureus. Minimum inhibitory concentration (MIC) values were determined using the tetrazolium microplate assay to quantitatively evaluate the antibacterial activity of the extracts. Diethyl ether extracts of F. benghalensis var. krishnae leaves exhibited the strongest antimicrobial activity against B. subtilis as compared with the standard drug ampicillin. Identification of the bioactive fraction in the leaf diethyl ether extract was performed using high-performance thin-layer chromatography (HPTLC)‒bioautography and analysis of separated fractions using high-resolution liquid chromatography (HR-LC) and mass spectrometry (MS). Our results revealed that the bioactive fractions of F. benghalensis var. krishnae plant contained 15 potential antimicrobial components. These components were then subjected to in silico absorption, distribution, metabolism, and excretion (ADME) and toxicological analysis, which narrowed them down to three lead compounds. Molecular docking was performed using the lead compounds against target proteins from B. subtilis as well as the pathogenic species, Bacillus anthracis, where 5,7,4′-tri-O-methylcatechin demonstrated its potential as an antimicrobial drug candidate. [ABSTRACT FROM AUTHOR]

Gallic acid is a phenolic organic acid found in plants that has a wide range of applications, as a health-promoting agent and as a food preservative, due to its high antioxidant activity. Gallic acid can be obtained by acid hydrolysis of tannic acid or by the enzymatic action of tannase on tannic acid. A biorefinery approach to produce gallic acid was developed. A combination of grape pomace and soybean hull as support substrate was used for solid-state fermentation by Aspergillus niger and Aspergillus oryzae. The best conditions for the bioconversion of tannic acid to gallic acid were tested. Tannase and other relevant enzymes were produced in the same process. The gallic acid production was monitored and quantified by high-performance thin-layer chromatography (HPTLC). HPTLC was used to separate the phenolic extracts prepared, and bioautography by gel overlay was used to identify antioxidants and tyrosinase inhibitors directly on the plate. The best gallic acid producer was A. oryzae, and the best fermentation condition comprised soybean hull and grape pomace as support substrate, producing 0.36 g of gallic acid/g of tannic acid and 7.2 g/L of fermentation medium in 72 h of incubation. The value addition of these agricultural wastes through a green process to produce gallic acid was demonstrated, creating opportunities for solid waste usage. [ABSTRACT FROM AUTHOR]

The smoke produced from natural substances such as medicinal plants is used in various cultures for different purposes. The use of medicinal fumes has been reported in nearly 50 countries. Among medicinal plants, Tussilago farfara L. known as coltsfoot has been introduced in Canon the famous book of Avicenna a Persian polymath, for chronic dry cough and various pulmonary diseases and shortness of breath. T. farfara is distributed in wet mountainous regions of Iran. For this study, the leaves and flowers of T. farfara were collected from Chalous Road in Iran. The smoke from the burning of T. farfara organs was prepared by homemade glassware trapping the smoke in methanol and then methanol was evaporated. In general, five grams of materials were burned and the smoke was dissolved and trapped in 100 ml of methanol. The trapped and dried materials from the smoke of extracts were filtered and injected into the GC/MS for analysis and identification of its constituents. 51 compounds representing 91.1 and 92.3 percent of smoke extracts of T. farfara were identified in leaf and flower. Also, 57 compounds were detected in the sample of EL and EF with 96.8 % and 97.7 %. The percentage of phenolic compounds that were identified in all extracts of smoke were SL and SF with 52.1 and 46.5, respectively. Phenol, Hydroquinone, P-Cresol and O-Cresol were the major compounds in the smoke extracts. Smoke leaves and flowers of T. farfara were selected to test the antimicrobial to continue. This study examined the bactericidal effect of smoke flowers. Fractions of effective constituents with the help of hexane-ethyl acetate with the method of thin layer chromatography (TLC) were isolated. The results of this experiment showed that a fraction (8:2) of hexane-ethyl acetate inhibited the bacteria Staphylococcus aureus. But Escherichia coli was not inhibited.

Poplars provide medicinal raw plant materials used in pharmacy. Leaf buds are one of the herbal medicinal products collected from poplars, having anti-inflammatory and antiseptic properties, but there are no quality standards for their production and there is a need to determine their botanical sources. Therefore, the chemical compositions of the leaf buds from four species and varieties of poplars, Populus balsamifera, P. × berolinensis, P. × canadensis ‘Marilandica’, and P. wilsonii were investigated and compared using gas chromatography coupled with mass detection (GC-MS) and two-dimensional high-performance thin-layer chromatography (2D-HPTLC) in order to search for taxa characterized by a high content of biologically active compounds and with a diverse chemical composition that determines their therapeutic effects. The presence of 163 compounds belonging to the groups of flavonoids, phenolic acids derivatives, glycerides, and sesquiterpenes was revealed. Moreover, the conditions for the separation and identification of biologically active compounds occurring in analyzed leaf buds using 2D-HPTLC were optimized and used for metabolomic profiling of the studied poplars, enabling their fast and simple botanical identification. The total phenolic (TPC) and flavonoid (TFC) contents of examined extracts were determined and their antioxidant capacities were estimated by spectrophotometric DPPH, ABTS, and FRAP assays. Based on the analysis of phytochemicals and antioxidant activity, P. × berolinensis buds were selected as the raw plant material for medicinal purposes with the highest content of active compounds and the strongest antioxidant activity.

Objectives: The ecological niche of soil for microorganisms that produce beneficial naturally occurring and biologically active products suitable for pharmaceutical industries is widely recognized. The purpose of the current study is to isolate fungus from soil samples of Dandeli, Karnataka and its metabolic profiling and investigating biological activities. Methods: Soil fungal isolates were screened for their antimicrobial potency against pathogens. The selected fungal isolate was morphologically characterized and ascertained by 18S rRNA gene sequencing, and the extract was investigated by gas chromatography-mass spectrometry (GC–MS) to identify the bio-molecules. Later the fungal extract was evaluated for antioxidant, antimicrobial, anti-mycobacterial and anti-proliferative activity against Human acute leukemia (K562) cell line. Results: The potential fungal isolate, Alternaria sp. KUDB15 showed a number of bioactive compounds through GC–MS analysis. A significant antioxidant activity (IC50 = 37.60 µg/ml) was also reported for this organism. A significant inhibitory activity against tested pathogens was shown by Alternaria sp. KUDB15 along with a strong anti-mycobacterial activity with an IC50 value of 3.12 μg/ml. During anticancer assay, this organism showed a concentration dependent reduction in cell viability having IC50 of 112.09 µg/ml. Further purification of the crude extract with column chromatography and thin layer chromatography (TLC) recorded several bands and the particular band having antibacterial activity was determined by bioautography. Conclusions: In the present research, the in vitro evaluation of biological activities of the fungal extract suggested that it might serve as a useful foundation for the discovery of compounds that have potential for medical applications. Additional research is required to identify and understand the structure of bio-molecules.

Background: Sphaeranthus indicus L. is a well-known medicinal plant in folk medicine. A variety of biological activities and chemical substances in this plant have been reported. The phytochemical content and activity may vary according to geographic location. This study aims to determine the geographical significance, the analysis of the synergistic effect of phytochemicals, the identification of active compounds, and the determination of the action mechanism of S. indicus inflorescence methanolic extract against Staphylococcus aureus and Klebsiella pneumonia. Results: The bands with Rf values of 0.92 and 1.0 showed antimicrobial activity, while all bands showed antioxidant activity. The first fraction showed the highest antimicrobial activity, and the pool of the second fraction showed the highest antioxidant activity. The kinetics of the antioxidants differed among the fractions. Analysis of synergistic effects showed that several compounds were involved in the activities. The bands with Rf 0.45, 0.55, 0.68, 0.79, and 0.85 were active components of the extract. Leakage of cell contents was detected at 260 and 280 nm wavelengths. Six different proteins and one nucleic acid band were detected after electrophoresis. The SEM analysis showed that the phytochemicals caused severe membrane damage. Conclusion: The study revealed that the photochemical present in methanol extract of the inflorescence of S. indicus has a synergistic effect and acts on bacterial cell envelope. The five compounds were identified as active molecules belonging to the class of terpenoids. The result also signified the geographical area since thymol was identified for the first time in this plant at this location. [ABSTRACT FROM AUTHOR]

Because of the increase in bacterial multidrug resistance, an ever-growing interest in the antimicrobial properties of plant extracts and plant-derived compounds. This necessitates the optimization of screening methods used for the identification of antimicrobials from other natural sources is of great importance. In this book chapter, an exhaustive list of in vitro antimicrobial susceptibility testing (AST) methods and detailed information on their advantages and limitations are reported. A highlight of the interpretation of the antibacterial activity of plant extracts of pure plant compounds is also discussed. The currently available screening methods for the detection of antimicrobial activity of natural products are classified into three main groups, including diffusion, bioautographic, and dilution methods. Therefore, others such as ATP bioluminescence assay, microfluidic-based culture, and flow cytofluorometric methods are not widely used because they require specified equipment and further evaluation for reproducibility and standardization. Testing methods provide reliable results if they follow standard procedures. Ideally, test systems should be simple, rapid, reproducible, inexpensive, and maximize sample throughput. Finally, the broth-microdilution for screening antibacterials from a plant source remains appropriate. [ABSTRACT FROM AUTHOR]

The smoke produced from natural substances such as medicinal plants is used in various cultures for different purposes. The use of medicinal fumes has been reported in nearly 50 countries. Among medicinal plants, Tussilago farfara L. known as coltsfoot has been introduced in Canon the famous book of Avicenna a Persian polymath, for chronic dry cough and various pulmonary diseases and shortness of breath. T. farfara is distributed in wet mountainous regions of Iran. For this study, the leaves and flowers of T. farfara were collected from Chalous Road in Iran. The smoke from the burning of T. farfara organs was prepared by homemade glassware trapping the smoke in methanol and then methanol was evaporated. In general, five grams of materials were burned and the smoke was dissolved and trapped in 100 ml of methanol. The trapped and dried materials from the smoke of extracts were filtered and injected into the GC/MS for analysis and identification of its constituents. 51 compounds representing 91.1 and 92.3 percent of smoke extracts of T. farfara were identified in leaf and flower. Also, 57 compounds were detected in the sample of EL and EF with 96.8 % and 97.7 %. The percentage of phenolic compounds that were identified in all extracts of smoke were SL and SF with 52.1 and 46.5, respectively. Phenol, Hydroquinone, P-Cresol and O-Cresol were the major compounds in the smoke extracts. Smoke leaves and flowers of T. farfara were selected to test the antimicrobial to continue. This study examined the bactericidal effect of smoke flowers. Fractions of effective constituents with the help of hexane-ethyl acetate with the method of thin layer chromatography (TLC) were isolated. The results of this experiment showed that a fraction (8:2) of hexane-ethyl acetate inhibited the bacteria Staphylococcus aureus. But Escherichia coli was not inhibited. [ABSTRACT FROM AUTHOR]

IntroductionThe need for new tools to treat infections is constantly growing due to the possibilities of emerging diseases related to environmental changes, climatic catastrophes, microorganism resistance, and human and animal aging, leading to an evident unbalance in the planet’s health. Brazil contains the most significant portion of world biodiversity, a potential source of new antimicrobial natural products. Nonetheless, its environment, particularly its forests, and rainforests, is under threat, meaning that rapidly conducted, comprehensive research into the potential of antimicrobial activity to address this threat is urgently needed.MethodsIn this study, plants from the Amazon rainforest and the Atlantic forests were collected and tested against several pathogenic microbes relevant to humans, animals, and the environment, and subjected to large-scale susceptibility assays, bioautography, and Artemia salina toxicity assays. From the plants, 2,280 organic and aqueous extracts were obtained from different organs, namely leaves, barks, flowers, fruits, and seeds, and subjected to a large-scale susceptibility screening assay against Staphylococcus aureus, Staphylococcus epidermidis, Enterococcus faecalis, Streptococcus mutans, Streptococcus sanguinis, Escherichia coli, Pseudomonas aeruginosa, Candida albicans, Malassezia pachydermatis, Malassezia furfur, and Listeria monocytogenes. Results and discussionThe selected extracts were subjected to antimicrobial susceptibility tests to determine their inhibition zone diameters and minimum bactericidal concentrations, to bioautography, and to an Artemia salina toxicity assay, which resulted in 154 active extracts. Moreover, 111 out of 154 extracts were ranked based on scores established by the p-values and the mean rank differences in each set of test results. The final ranking identified which extracts should be studied in further phytochemical research using thin-layer chromatography techniques as a priority. The extracts obtained from plants belonging to Combretaceae, Connaraceae, Convolvulaceae, Fabaceae, Malpighiaceae, Moraceae, Piperaceae, Polygonaceae, and Salicaceae were selected as the most promising ones and used to support the identification of plant-based antimicrobial active compounds from the immense biodiversity of Brazilian forests.

Moringa oleifera, native to India, grows in tropical and subtropical regions around the world and has valuable pharmacological properties such as anti-asthmatic, anti-diabetic, anti-inflammatory, anti-infertility, anti-cancer, anti-microbial, antioxidant, and many more. The purpose of this study was to assess the free radical scavenging ability of two extracts and two pure compounds of M. oleifera Lam (hexane, ethanol, compound E3, and compound Ra) against reactive oxygen species, as well as their reducing power and antimicrobial activities. Bioautography antioxidant assay, 2,2-diphenyl-1-picrylhydrazyl (DPPH), hydrogen peroxide (H2O2) free radical scavenging, and iron (iii) (Fe3+ to Fe2+) chloride reducing power assays were used to assess the extracts' qualitative and quantitative free radical scavenging activities. Furthermore, the extract and the compounds were tested against both Gram-positive and Gram-negative bacterial strains suspended in Mueller–Hinton Broth. The extracts and pure compounds showed noteworthy antioxidant potential, with positive compound bands in the Rf range of 0.05–0.89. DPPH), H2O2, and Fe3+ to Fe2+ reduction assays revealed that ethanol extract has a high antioxidant potential, followed by compound E3, compound Ra, and finally hexane extract. Using regression analysis, the half maximal inhibitory concentration (IC50) values for test and control samples were calculated. Compound Ra and ethanol exhibited high antioxidant activity at concentrations as low as ≈ 0.28 mg/mL in comparison with n-hexane extract, compound E3, ascorbic acid, and butylated hydroxytoluene standards. The radical scavenging activity of almost all M. oleifera plant extracts against DPPH was observed at 0.28 mg/mL; however, the highest activity was observed at the same concentration for ascorbic acid and butylated hydroxytoluene (BHT) with a low IC50 value of 0.08 mg/mL and compound Ra and ethanol with a low IC50 of 0.4 mg/mL, respectively. The extracts and pure compounds of M. oleifera have little to no antibacterial potential. M. oleifera extracts contain antioxidant agents efficient to alleviate degenerative conditions such as cancer and cardiovascular disease but have little activity against infectious diseases. [ABSTRACT FROM AUTHOR]

Introduction: Many species within Combretaceae are traditionally used for the treatment of bacterial infections. The similarity in chemistry and antimicrobial activities within the family pose a challenge in selecting suitable species for herbal drug development. Objective: This study aimed at rapidly identifying antimicrobial compounds using bioautography‐guided high‐performance thin‐layer chromatography coupled with mass spectrometry (HPTLC–MS). Methods: Hierarchical cluster analysis of ultra‐performance liquid chromatography‐mass spectrometry data from the methanol extracts of 77 samples, representing four genera within Combretaceae, was carried out. Based on groupings on the dendrogram, 15 samples were selected for bioautography analysis against four pathogens (Staphylococcus aureus, Bacillus cereus, Escherichia coli and Salmonella typhimurium). Active compounds were identified using HPTLC–MS analysis of bands corresponding to the inhibition zones. Results: Bioautography revealed 15 inhibition zones against the four pathogens, with the most prominent present for Combretum imberbe. Analysis of the active bands, using HPTLC–MS indicated that flavonoids, triterpenoids and combretastatin B5 contributed to the antibacterial activity. The compounds corresponding to molecular ions m/z 471 (Combretum imberbe) and 499 (Combretum elaeagnoides) inhibited all four pathogens, and were identified as imberbic acid and jessic acid, respectively. Chemotaxonomic analysis indicated that arjunic acid, ursolic acid and an unidentified triterpenoid (m/z 471) were ubiquitous in the Combretaceae species and could be responsible for their antibacterial activities. Conclusion: Application of HPTLC–MS enabled the rapid screening of extracts to identify active compounds within taxonomically related species. This approach allows for greater efficiency in the natural product research workflow to identify bioactive compounds in crude extracts. This study was aimed at rapidly identifying antimicrobial compounds within Combretaceae using bioautography‐guided high‐performance thin‐layer chromatography‐mass spectrometry (HPTLC‐MS). Representative samples were subjected to bioautography against bacterial pathogens, and compounds were identified from the active bands using HPTLC‐MS. Chemotaxonomic analysis revealed that arjunic acid, ursolic acid, and an incompletely identified triterpenoid (m/z 471) were ubiquitous in the Combretaceae species and associated with antibacterial activity. This approach enables rapid extract screening to identify active compounds within taxonomically related species. [ABSTRACT FROM AUTHOR]

Natural products and their analogues have contributed significantly to treatment options, especially for anti-inflammatory and infectious diseases. Thus, the primary objective of this work was to compare the bioactivity profiles of selected medicinal plants that are historically used in folk medicine to treat inflammation and infections in the body. Chemical HPTLC fingerprinting was used to assess antioxidant, phenolic and flavonoid content, while bioassay-guided HPTLC was used to detect compounds with the highest antibacterial and anti-inflammatory activities. The results of this study showed that green tea leaf, walnut leaf, St. John’s wort herb, wild thyme herb, European goldenrod herb, chamomile flower, and immortelle flower extracts were strong radical scavengers. Green tea and nettle extracts were the most active extracts against E. coli, while calendula flower extract showed significant potency against S. aureus. Furthermore, green tea, greater celandine, and fumitory extracts exhibited pronounced potential in suppressing COX-1 activity. The bioactive compounds from the green tea extract, as the most bioactive, were isolated by preparative thin-layer chromatography and characterized with their FTIR spectra. Although earlier studies have related green tea’s anti-inflammatory properties to the presence of catechins, particularly epigallocatechin-3-gallate, the FTIR spectrum of the compound from the most intense bioactive zone showed the strongest anti-inflammatory activity can be attributed to amino acids and heterocyclic compounds. As expected, antibacterial activity in extracts was related to fatty acids and monoglycerides.

Quality control of herbal medicines is an important stage in generating effective and safe herbal remedies. A unique high-throughput approach for assessing anti-diabetic phytoconstituents in Syzygium cumini seed extract and marketed formulation was examined using high-performance thin-layer chromatography (HPTLC)–bioautography technology. Using β-sitosterol as reference, this agar-free bioautographic technique was devised and validated for the assessment of α-amylase inhibitors on TLC plates. The presence of some anti-diabetic compounds in plant samples was verified using a combination of biological detection and mass spectrometry (HPTLC–MS). The newly proposed HPTLC–bioautography screening method may be effective for evaluating the anti-diabetic activity of plant extracts and formulations. The proposed technique is quick, easy, precise, and dependable. [ABSTRACT FROM AUTHOR]

The current study is aimed to isolate endophytic actinobacteria from various medicinal plants for exploring its antimicrobial activity against clinical pathogens. Out of 50 actinobacterial cultures isolated from 11 medicinal plants, 58 % of the cultures showed antimicrobial activity against at least two out of nine pathogens tested. Strain KCA1 isolated from Phyllanthus niruri leaves showed maximum zone of inhibition against all of the nine pathogens tested. Strain KCA1 exhibited maximum level of antimicrobial metabolite production in solid state fermentation during 9th d of incubation. Three well separated spots were observed when the ethyl acetate extract of strain KCA1 was analyzed through thin layer chromatography. One spot with retention factor value 0.82 was found to inhibit Staphylococcus aureus in thin layer chromatography bio autographic assay. The active fraction eluted from thin layer chromatography was further characterized by gas chromatography-mass spectroscopy analysis. The mass spectral data revealed the presence of three compounds viz. 2, 4-di-tert-butylphenol, 1-hexadecanol and 1-nonadecene present in major proportions which are responsible for the antimicrobial activity of the strain KCA1. In fermentation experiment, variables such as glucose, yeast extract and sodium chloride were found to influence the antimicrobial compound production. The potential strain KCA1 was identified as Streptomyces sp. on the basis of microscopic, cultural, physiological and 16S ribosomal ribonucleic acid analysis. [ABSTRACT FROM AUTHOR]

Salvia africana-lutea L., S. lanceolata L., and S. chamelaeagnea L. are used in South Africa as traditional medicines to treat infections. This paper describes an in-depth investigation into their antibacterial activities to identify bioactive compounds. Methanol extracts from 81 samples were screened against seven bacterial pathogens, using the microdilution assay. Biochemometric models were constructed using data derived from minimum inhibitory concentration (MIC) and ultra-performance liquid chromatography-mass spectrometry data. Active molecules in selected extracts were tentatively identified using high-performance thin layer chromatography (HPTLC), combined with bioautography, and finally, by analysis of active zone eluates by mass spectrometry (MS) via a dedicated interface. Salvia chamelaeagnea displayed notable activity towards all seven pathogens, and the activity, reflected by MICs, was superior to that of the other two species, as confirmed through ANOVA. Biochemometric models highlighted potentially bioactive compounds, including rosmanol methyl ether, epiisorosmanol methyl ether and carnosic acid. Bioautography assays revealed inhibition zones against A. baumannii, an increasingly multidrug-resistant pathogen. Mass spectral data of the eluted zones correlated to those revealed through biochemometric analysis. The study demonstrates the application of a biochemometric approach, bioautography, and direct MS analysis as useful tools for the rapid identification of bioactive constituents in plant extracts. [ABSTRACT FROM AUTHOR]

Background: Sea buckthorn (Hippophae) is in the focus of interest mainly for its positive effects on health of both human and animal organisms. Due to the similarities in vegetative morphology, Hippophae species are often misidentified. Therefore, current study was focused on ITS based sequence characterization of sea buckthorn species and comparative biochemical evaluation for its antioxidant properties. Methods and results: DNA was extracted from leaf samples. Primer pairs K-Lab-SeaBukRhm-ITS1F1- K-Lab-SeaBukRhm-ITS1R1 and K-LabSeaBukTib- ITSF1- K-LabSeaBukTib-ITSR1 were used for PCR amplification. The purified PCR products were outsourced for sequencing. Phylogenetic tree was constructed based on neighbor-joining (NJ) method. Moreover, comparison of antioxidant potential of leaves of two sea buckthorn species (Hippophae rhamnoides and Hippophae tibetana) collected from different regions of Ladakh viz., Stakna, Nubra, DRDO Leh and Zanskar was determined by 2,2-diphenyl-1-picrylhydrazyl (DPPH), 2,2-azino-bis (3- ethylbenzothiazoline-6-sulphonic acid) diammonium salt (ABTS), and Total antioxidant capacity (TAC) by phosphomolybdenum assays. The present investigation led to the differentiation of two sea buckthorn species viz., H. rhamnoides and H. tibetana based on Internal Transcribed Spacer (ITS) region. Moreover, significant variation was observed in antioxidant potential of leaf extracts collected from different regions. Conclusions: Primary ITS sequence analysis was found to be powerful tool for identification and genetic diversity studies in sea buckthorn. Leaves of sea buckthorn have pronounced antioxidant properties and can be used in food, neutraceuticals and pharmaceutical industries etc. The current study will pave the way to discover small bioactive molecules responsible for antioxidant and anticancer properties in sea buckthorn. [ABSTRACT FROM AUTHOR]

[Display omitted] • Thin layer chromatography assay developed for laccase inhibitor detection. • Optimization of on-plate laccase reaction with substrate for enhanced contrast. • Integration of control assay on TLC distinguishes true inhibitors from scavengers. • Detection and quantification limits for sodium azide imply good sensitivity. • Successful application of the method to phenolic and non-phenolic hydrazones. Thin-layer chromatography (TLC) hyphenated to bioassays is a modern tool used for discovery of biologically active compounds from complex mixtures. The first bioautographic assay for detecting laccase inhibitors on a TLC plate was developed in this study. The on-plate reaction of laccase with colourless ABTS that renders the blue ABTS∙+ radical was optimised. Combination of the enzymatic TLC-assay with a control TLC-assay, wherein ABTS∙+ radical is chemically generated and then applied on the TLC, allowed to differentiate between the pure laccase inhibitor sodium azide and radical scavengers such as gallic and kojic acids. The limit of detection and quantification for the method were 54.9 and 166 ng of sodium azide respectively. The methodology was applied successfully to a recently discovered laccase inhibitor chemotype: hydrazones. A model hydrazone was compared with several hydrazones synthesized for this study. For the first time, laccase inhibitors separated on a TLC plate can be detected individually. [ABSTRACT FROM AUTHOR]

The current investigation aims to test the susceptibility of human pathogenic clinical isolates and MTCC strains to leaf and seed extracts of Terminalia catappa and Nigella sativa. Disc diffusion assay, micro dilution assay and minimum Bactericidal Concentration investigated the susceptibility of bacteria to the test extracts. The active extract was subjected to phytochemical screening, separation of the phytochemicals by Thin Layer Chromatography, bioactivity guided assay and Time- kill assay. Acetone and methanol extracts of T.catappa revealed, significant inhibition of clinical origin Staphylococcus aureus followed by Proteus vulgaris and the MTCC strains Staphylococcus aureus, Salmonella typhi, Pseudomonas aeroginosa and Bacillus subtilis. Nigella sativa inhibited the growth of clinical origin Staph.aureus and MTCC strain of Staph.aureus, Salmonella typhi and B.subtilis. Minimum inhibitory concentration for all the test bacteria was reported in the range of 5000μg/ml to 9 μg/ml in T. catappa extract. Most sensitive being the clinical isolate Staph. aureus and Proteus vulgaris. The bactericidal concentration for the test bacteria was found to be between 5000μg/ml and 625μg/ml. Phyto-chemical analysis of leaf extracts of T. catappa found to have dominated by polyphenols (Terpenoids, steroids, flavonoids, flavones, saponins and tannins) and N.sativa extracts recorded the presence of alkaloids, proteins and oils and fats. TLC profiling of the acetone extract revealed many antibacterial active bands. Bands having Retention factor 0.47 and 0.52 were active against the test bacteria. Time kill assay of the acetone extract of T. catappa were carried out for the first time. The extract exhibited dose dependent bactericidal and bacteriostatic activity against the clinical isolates.

In the present study AIA25a strain has been isolated from the rhizospheric soil actinomycetes from selected locations of Rajasthan. Antimicrobial activity test of AIA25a against bacterial strains S. aureus (MTCC 3160), P. aeruginosa (MTCC 1688), P. mirabilis (ITCC 425) has been carried out. A good activity is shown by chloroform crude extract against P. aeruginosa (IZ=13.4±0.3mm), moderate activity (IZ=11.1±0.1) is shown against S. aureus and no activity is observed against P. mirabilis strain. Different compounds isolation is executed with the help of chloroform solvent. Using GC-MS identification has been carried out for compounds present in AIA25a strain followed by partial purification of compounds. With the help of HPTLC method partial purification of compounds has been achieved and followed by GC-MS analysis for identification of separated compounds. The Bioautography technique has been applied for the detection of antimicrobial spot of partial purified compounds of crude extract of AIA25a isolate. Results of this study show the presence of various compounds in the rhizospheric soil of Rajasthan. [ABSTRACT FROM AUTHOR]

Followed by a buildup of its phytochemical profile, Erodium cicutarium is being subjected to antimicrobial investigation guided with its ethnobotanical use. The results of performed in vitro screening on Staphylococcus aureus, Pseudomonas aeruginosa, and Candida albicans strains, show that E. cicutarium has antimicrobial activity, with a particular emphasis on clinical S. aureus strains—both the methicillin sensitive (MSSA) and the methicillin resistant (MRSA) S. aureus. Experimental design consisted of general methods (the serial microdilution broth assay and the agar well diffusion assay), as well as observing bactericidal/bacteriostatic activity through time (the "time-kill" assay), investigating the effect on cell wall integrity and biofilm formation, and modulation of bacterial hemolysis. Observed antibacterial activity from above-described methods led to further activity-guided fractionation of water and methanol extracts using bioautography coupled with UHPLC-LTQ OrbiTrap MS4. It was determined that active fractions are predominantly formed by gallic acid derivatives and flavonol glycosides. Among the most active phytochemicals, galloyl-shikimic acid was identified as the most abundant compound. These results point to a direct connection between galloyl-shikimic acid and the observed E. cicutarium antibacterial activity, and open several new research approaches for future investigation. [ABSTRACT FROM AUTHOR]

Moringa oleifera, native to India, grows in tropical and subtropical regions around the world and has valuable pharmacological properties such as anti-asthmatic, anti-diabetic, anti-inflammatory, anti-infertility, anti-cancer, anti-microbial, antioxidant, and many more. The purpose of this study was to assess the free radical scavenging ability of two extracts and two pure compounds of M. oleifera Lam (hexane, ethanol, compound E3, and compound Ra) against reactive oxygen species, as well as their reducing power and antimicrobial activities. Bioautography antioxidant assay, 2,2-diphenyl-1-picrylhydrazyl (DPPH), hydrogen peroxide (H2O2) free radical scavenging, and iron (iii) (Fe3+ to Fe2+) chloride reducing power assays were used to assess the extracts’ qualitative and quantitative free radical scavenging activities. Furthermore, the extract and the compounds were tested against both Gram-positive and Gram-negative bacterial strains suspended in Mueller–Hinton Broth. The extracts and pure compounds showed noteworthy antioxidant potential, with positive compound bands in the Rf range of 0.05–0.89. DPPH), H2O2, and Fe3+ to Fe2+ reduction assays revealed that ethanol extract has a high antioxidant potential, followed by compound E3, compound Ra, and finally hexane extract. Using regression analysis, the half maximal inhibitory concentration (IC50) values for test and control samples were calculated. Compound Ra and ethanol exhibited high antioxidant activity at concentrations as low as ≈0.28 mg/mL in comparison with n-hexane extract, compound E3, ascorbic acid, and butylated hydroxytoluene standards. The radical scavenging activity of almost all M. oleifera plant extracts against DPPH was observed at 0.28 mg/mL; however, the highest activity was observed at the same concentration for ascorbic acid and butylated hydroxytoluene (BHT) with a low IC50 value of 0.08 mg/mL and compound Ra and ethanol with a low IC50 of 0.4 mg/mL, respectively. The extracts and pure compounds of M. oleifera have little to no antibacterial potential. M. oleifera extracts contain antioxidant agents efficient to alleviate degenerative conditions such as cancer and cardiovascular disease but have little activity against infectious diseases.

Hydrocotyle javanica Thunb. (Family Araliaceae) has been used in traditional herbal medicine as a substitute of Centella asiatica (L.) Urban. Although, the latter has been studied extensively and various biologically active constituents have been reported, the former has not been studied for its active constituents. The current study followed activity guided isolation and identification of the fraction with antimicrobial activity from H. javanica. Plant extracts were prepared by extracting successively with petroleum ether, ethyl acetate and ethanol. Antimicrobial activity was evaluated using gram positive and gram negative bacteria and Candida albicans. Bioactivity-guided isolation was carried out by preparative thin layer chromatography and bioautography. Cytotoxicity of the most active antimicrobial principle was evaluated against Human embryonic kidney (HEK293) and Adenocarcinomic Human Alveolar Basal Epithelial (A549) cell lines. The tentative structure of the most active antimicrobial principle was constructed from the data obtained from FTIR, MS, H-NMR and C-NMR. [ABSTRACT FROM AUTHOR]

Abstract Resistant bacterial infections are a major public health problem worldwide, which entails the need to search for new therapeutic agents. In this context, lichens stand out, provided that they are producers of structurally diverse compounds that have attractive biological properties, including antimicrobial activity. Thus, extracts of 12 lichen species were prepared and their potential to inhibit the growth of 5 bacterial strains was evaluated in this work. The chemical compositions of these extracts were examined using TLC and microcrystallization, being the identity of the active compounds in each extract attributed based on the bioautography technique. The most active extracts (and their identified active compounds) were from Cladonia borealis (usnic, barbatic and 4-O-demethylbarbatic acids), Cladina confusa (usnic and perlatolic acids), Stereocaulom ramulosum (atranorin, perlatolic and anziaic acids) and Canoparmelia cryptochlorophaea (cryptochlorophaeic and caperatic acids), with MICs ranging from 7.8 to 31.25 μg/mL, including for resistant clinical strains. MIC values were also obtained for substances isolated from lichens for comparison purposes. A group of four extracts containing usnic acid was analyzed by 1H NMR in order to correlate relative proportion of major metabolites and extracts activity. The less active extracts in this group, in fact, presented low proportion of usnic acid.

The intriguing discovery of antibiotics that led to innumerable accomplishments in the past is slowly losing its efficiency in the clinical society. The emergence of antibiotic-resistant pathogens, such as those which can hydrolyze β-lactam antibiotics with the help of enzymes such as Extended Spectrum β-lactamases (ESBLs) has created an urgent necessity to witness innovation in the current drug therapy, and develop new antimicrobial agents. Keeping the current clinical adversity in mind, the present study was carried out to investigate the antibacterial activity of Pimenta dioica (L.) Merr. extracts, prepared in various chemical solvents with the help of soxhlet apparatus, and its synergistic activity with ampicillin. The antibacterial activity of these extracts was verified against 45 ESBL producing bacteria. Our study revealed chloroform extract to exhibit maximum antibacterial activity evident with the zones of inhibition (12-17 mm) showed by agar diffusion method, and their observed Minimum Bactericidal Concentrations (MBC) in the range of 2-5 mg.ml-1. The MBC of ampicillin was found to decrease from 10 mg.ml-1 to 300-500 μg.ml-1 in presence of chloroform extracts of P. dioica (L.) Merr. leaves, indicating a synergistic activity between them. In addition, the TLC bioautography of chloroform extract was carried out using 2, 3, 5-Trimethyl tetrazolium chloride as a chromogen, and the separated bioactive components were analyzed with the help of GC-MS which showed eugenol as a major constituent. Moreover, the scanning electron microscope analysis confirmed a visible deformation in the cell membrane of E. coli treated with chloroform extracts of P. dioica (L.) Merr., indicating stress and cellular damage in presence of these extracts. [ABSTRACT FROM AUTHOR]

• TLC bioautography directed approach from Nigrospora sphaerica revealed antimicrobial nigrosporalactone. • Nigrosporalactone exhibited broad-spectrum activity against human pathogens. • Docking studies revealed bioactive could act on a wide range of targets in pathogens. TLC-bioautography amalgamated with hyphenated spectroscopy aid in precise in situ detection of secondary metabolites with pharmaceutical significance. TLC bioautography offers efficient and economical strategy in identifying compounds of interest from crude extracts. The present investigation has been focused on detection of antimicrobial metabolite from the culture broth of Nigrospora sphaerica inhabiting Croton bonplandianus Baill. The antimicrobial profiling confirmed the bioactive nigrosporalactone to possess broad-spectrum activity against test human pathogens with minimum inhibitory concentration values in the range 6.25 µg to 100 µg. The in silico studies revealed protein targets 1I01 (E. coli beta-ketoacyl reductase), 1IYL (C. albicans N-myristoyl transferase) had the highest binding score of -6.1 Kcal/mol. [ABSTRACT FROM AUTHOR]

Vegetable tars are newly formed resinous products extracted by pyrolysis from wood or bark. Since prehistoric times, they have been used for their adhesive, water-repellent, repellent and finally medicinal properties. This work investigated the use of these vegetable tars as sources of biofungicides. The physicochemical properties of the tars obtained in the laboratories showed a significant yield of 1.22%, 0.66% and 0.51% respectively for the wood of (Jeniperus phoenicea, Olea europeae sylvestris and Acacia tortilis Sub raddiana). They also showed a density value of 1.22 and a refractive index of about 1.5115 for J.phoenicea, an acid number of 0.39 and a dry matter ratio of 19.86% for A. tortilis Sub raddiana. Considerable antifungal activity of the acetate of three tars against certain fungal phytopathogens (Fusarium) was observed with the most remarkable MIC values ranging from 0.1 mg / ml to 0.46 mg / ml. The J. phoenicea extracts were more active against F. oxysporum f sp albedinis (FOA) strains. The growth of F. solani was more sensitive to acetate extracts from O. europeae. Dichloromethane extracts of A. tortilis Sub raddiana yielded with an average MIC of 0.16 and 1.02 mg / ml, respectively for strains of F. oxysporum and F. graminearum. The bioautography test revealed the presence of 3 antifusaric molecules of interest. These MIC results further merit in vivo tests for the development of future biofungicides. [ABSTRACT FROM AUTHOR]

Salvia africana-lutea L., S. lanceolata L., and S. chamelaeagnea L. are used in South Africa as traditional medicines to treat infections. This paper describes an in-depth investigation into their antibacterial activities to identify bioactive compounds. Methanol extracts from 81 samples were screened against seven bacterial pathogens, using the microdilution assay. Biochemometric models were constructed using data derived from minimum inhibitory concentration (MIC) and ultra-performance liquid chromatography-mass spectrometry data. Active molecules in selected extracts were tentatively identified using high-performance thin layer chromatography (HPTLC), combined with bioautography, and finally, by analysis of active zone eluates by mass spectrometry (MS) via a dedicated interface. Salvia chamelaeagnea displayed notable activity towards all seven pathogens, and the activity, reflected by MICs, was superior to that of the other two species, as confirmed through ANOVA. Biochemometric models highlighted potentially bioactive compounds, including rosmanol methyl ether, epiisorosmanol methyl ether and carnosic acid. Bioautography assays revealed inhibition zones against A. baumannii, an increasingly multidrug-resistant pathogen. Mass spectral data of the eluted zones correlated to those revealed through biochemometric analysis. The study demonstrates the application of a biochemometric approach, bioautography, and direct MS analysis as useful tools for the rapid identification of bioactive constituents in plant extracts.

Suaeda monoica Forssk. ex J.F.Gmel. (Amaranthaceae), a mangrove associate and ethno-medicinal herb of Indian Sundarbans, was investigated as a promising source of bioactive compounds. Various polar and nonpolar solvent extracts of the leaf and root-shoot parts of the plant exhibited antioxidant, antibacterial, antifungal, allelopathic, mosquitocidal, antihaemolytic and antidiuretic potential. Moreover, to meet pharmacological requirements, the antioxidant ability of the plant was validated by both chemical and biological analyses. Extraction yield and presence of different phytochemicals like phenolics, flavonoids, tannins and saponins were compared in various solvent-extracted fractions. Principle component analysis revealed that the antioxidant property present in different extracts maintained a positive correlation with the occurrence of polyphenols (phenolics, tannins and flavonoids). Biochemical evaluation, HPLC examination and GC–MS analysis showed a differential level of the presence of various phytochemicals in different solvent extracts. In contrast to mosquitocidal, antioxidant, antihaemolytic and phytotoxic properties which were observed to be dominant in polar solvent extracts, maximum antibacterial potency was detected in nonpolar n-hexane fractions. Overall, the plant extract is nontoxic in nature and a dose amounting to 3,000 mg/kg was well tolerated by Swiss albino mice. A combination of HPLC and GC–MS analyses showed the presence of a large number of structurally diverse phytochemicals, many of which had already been reported as insecticidal, mosquitocidal, antibacterial, herbicidal, antidiuretic, antioxidant and anti-haemolytic compounds. All these findings support that the least explored traditional edible medicinal mangrove associate S.monoica is enriched with multiple bioactive molecules and may be considered as one of the richest sources of various lead molecules of pharmaceutical importance.

Followed by a buildup of its phytochemical profile, Erodium cicutarium is being subjected to antimicrobial investigation guided with its ethnobotanical use. The results of performed in vitro screening on Staphylococcus aureus, Pseudomonas aeruginosa, and Candida albicans strains, show that E. cicutarium has antimicrobial activity, with a particular emphasis on clinical S. aureus strains—both the methicillin sensitive (MSSA) and the methicillin resistant (MRSA) S. aureus. Experimental design consisted of general methods (the serial microdilution broth assay and the agar well diffusion assay), as well as observing bactericidal/bacteriostatic activity through time (the “time-kill” assay), investigating the effect on cell wall integrity and biofilm formation, and modulation of bacterial hemolysis. Observed antibacterial activity from above-described methods led to further activity-guided fractionation of water and methanol extracts using bioautography coupled with UHPLC-LTQ OrbiTrap MS4. It was determined that active fractions are predominantly formed by gallic acid derivatives and flavonol glycosides. Among the most active phytochemicals, galloyl-shikimic acid was identified as the most abundant compound. These results point to a direct connection between galloyl-shikimic acid and the observed E. cicutarium antibacterial activity, and open several new research approaches for future investigation.

Objectives: Carica papaya Linn. a member of the Caricaceae family, is a tropical fruit, rich in various secondary metabolites owing to its antioxidant, anti-inflammatory, anti-diabetic, and antihelmintic properties. The study was carried out to investigate the metabolic profiling and free radical scavenging compounds in C. papaya fruit's pulp, peel and seeds through thin-layer chromatography-mass spectrometry (TLC-MS) bioautography. Methodology: Aqueous, hydroalcoholic and alcoholic extracts of fruit pulp and peel were prepared along with the hydroalcoholic and hexane extract of seeds. These were subjected to total phenol, flavonoid and free radical scavenging estimation. Qualitative and quantitative high-performance thin-layer chromatography analysis of the best extracts was performed followed by TLC-MS bio autography assay for the detection of free radical scavenging compounds. Results: C. papaya peel was noted to contain the highest phenol and flavonoid content, but the seeds showed better free radical scavenging activity. The hydroalcoholic extracts of pulp, peel, and seeds examined through TLC-bioautography showed the presence of chlorogenic acid, ellagic acid, quercetin, ß-sitosterol, linoleic acid, and iso-oleic acid as potent free radical scavenging compounds. Liquid chromatography-mass spectrometry analysis showed the presence of ß-carotene, lycopene, and ß-cryptoxanthin in pulp and peel along with other carotenoids and benzyl isothiocyanate, linoleic acid, oleic acid, and methyl palmitate were the major compounds detected in seeds through gas chromatography-mass spectrometry analysis. Conclusion: This study has revealed that C. papaya fruit and seeds possess potent free radical scavenging compounds. Seeds which make up the waste material may be utilized in cosmetic industries as they signify rich antioxidants. [ABSTRACT FROM AUTHOR]

Jojoba (Simmondsia chinensis (Link)) is a shrub plant that widely used as cosmetic ingredients especially jojoba oil. When making jojoba oil, the residue still reminds and become waste. This study aimed to determine the antioxidant activity of jojoba seed residue (JSR). Jojoba seed residue was collected from Sudan. The JSR extracted by maceration with n-hexane, ethyl acetate, and 70% ethanol solvent. The antioxidant activity was determined using DPPH (2,2-diphenyl-1-picrylhydrazyl) assay. The result showed that 70% ethanol extract of JSR had the highest antioxidant activity with 50% Inhibition Concentration (IC50) value of 219.42 mg/L. The 70% ethanol extract was fractionated by using silica gel column chromatography with gradient elution produced 10 fractions. All fractions showed antioxidant activity (IC50 106-447 mg/L). Fraction 3 had the highest antioxidant activity with IC50 value of 106.26 mg/L. Therefore, JSR has a great potential for use as a raw material in cosmetic and pharmaceutical products industry, or as a source of bioactive compounds. [ABSTRACT FROM AUTHOR]

Acetone extracts of the two common epiphytes lichens Usnea florida and Flavoparmelia caperata have been evaluated for their antimicrobial activities against Staphylococcus aureus, Candida albicans and Aspergillus brasiliensis. The dibenzofuran derivative (+)-usnic acid (1) was the main metabolite in these two species. Thamnolic (5), evernic (6), physodic (7) and 3-hydroxyphysodic acids (8) were isolated from U. florida, as well as 5,7-dihydroxy-6-methylphtalide (2) which was newly identified in this Genus. Protocetraric (3) and caperatic acids (4) and ergosterol peroxide (9) are usually biosynthezised by F. caperata. Antibacterial activity was determined for the four main compounds against Staphylococcus aureus using bioautography and broth dilution method. Minimal inhibitory concentrations of usnic acid, caperatic acid and protocetraric acid were comprised between 7.25 and 12.5 µg/mL. [ABSTRACT FROM AUTHOR]

In this study, Limoniastrum feei extracts were evaluated for management of Fusarium oxysporum f. sp. albedinis (Foa) through antifungal tests and cellulases inhibition (cellulases are cell wall degrading enzymes “CWDE”). Antifungal activity evaluation was realized by direct bioautography. Cellulases inhibition test was realized by contact bioautography. In addition, a phytochemical screening has been done for these extracts on TLC plates (extraction with solvents of increasing polarity: hexane, ethyl acetate, dichloromethane, and methanol). Phytochemical screening was intended to know which metabolite is responsible for the antifungal activity through correlation with antifungal and anti-cellulases effect. The phytochemical study revealed the presence of saponins, flavonoids, tannins, coumarins, and alkaloids in these extracts. The different extracts showed high variability in their phenolic and flavonoids contents as well as antifungal capacity, as the case of ethyl acetate extract. Detection of inhibition zones was revealed with ethanolic Gram’s iodine. Inhibition zones were determined as dark brown spots. The results of enzymatic activity -by contact bioautography technique-showed that certain extracts of L. feei present a significant effect on cellulases of Foa. The best effect was presented by methanolic extract of aerial part (inhibition zone diameter in chromatogram is higher than 2 mm). Best antifungal effect was presented by compounds with high probability to be phenolic compounds.