Synergy based Extracts of Medicinal Plants: Future Antimicrobials to Combat Multidrug Resistance.

The use of herbal medicines and supplements in the last thirty years has increased enormously. Herbal medication has demonstrated promising and effective potential against various diseases. Herbal and phytoconstituent medications are gaining popularity globally and many people are adopting herbal remedies to deal with different health issues. The indiscriminate use of antibiotics, due to the development of antimicrobial resistance, poses an unprecedented problem for human civilization. Bacterial infections are difficult to cure because of the propensity of microbes to acquire resistance to a wide range of antimicrobial drugs. New compounds are being explored and quantified for possible antibacterial activity with little or no side effects. Researchers are investigating the range of therapeutic plants mentioned in Unani, Ayurveda, and Siddha around the globe. Known and commonly acclaimed global databases such as PubMed, Research Gate, Science Direct, Google Scholar were searched using different search strings such as Indian medicinal plants, multidrug resistance (MDR), thin layer chromatography (TLC), antimicrobials, and Synergism were used in diverse combinations to reclaim numerous citations associated with this area. Thus, the current review aims to shed a light on the information of medicinal plants as a potential foundation of herbal drugs and elucidate how synergism and TLC bioautography play a crucial role in finding antimicrobial compounds.

Phenolic and flavonoid contents and antioxidant activity of an endophytic fungus Nigrospora sphaerica (EHL2), inhabiting the medicinal plant Euphorbia hirta (dudhi) L.

Since endophytic fungi are pivotal sources of various bioactive natural compounds, the present study is aimed to investigate the antioxidant compounds of the endophytic fungus Nigrospora sphaerica isolated from a pantropical weed, Euphorbia hirta L. The fungus was fermented in four different media and each filtered broth was sequentially extracted in various solvents. Crude extracts collected from different solvents were subjected to phytochemical analysis and antioxidant activity. The total phenolic content (TPC) and total flavonoid content (TFC) were maximal in ethyl acetate crude extract (EtOAcE) of endophyte fermented in potato dextrose broth (PDB) medium (77.74 ± 0.046mgGAE/g and 230.59 ± 2.0 mgRE/g) with the highest 96.80% antioxidant activity. However, TPC and TFC were absent in hexane extract of Czapek Dox broth (CDB) medium exhibiting the lowest 4.63 ± 2.75% activity. The EtOAcE (PDB) showed a positive correlation between TFC and antiradical activity (R2 = 0.762; P < 0.05), whereas a high positive correlation was noticed between TPC and antioxidant activity (R2 = 0.989; P < 0.05). Furthermore, to determine the antioxidant activity, EtOAcE (PDB) was subjected to TLC bioautography-based partial purification, while GC/MS analysis of the partial purified extract was done to confirm the presence of phenolics along with antioxidant compounds that resulted in the detection of 2,4-Di-tert-butylphenol (13.83%), a phenolic compound accountable for the antioxidant potential. Conclusively, N. sphaerica is a potential candidate for natural antioxidant.

In-depth phytochemical and biological studies on potential AChE inhibitors in red and zigzag clover dry extracts using reversed-phase liquid chromatography (RP-LC) coupled with photodiode array (PDA) and electron spray ionization-quadrupole/time of flight-mass spectrometric (ESI-QToF/MS-MS) detection and thin-layer chromatography-bioautography.

In the study, Trifolium medium L. and T. pratense L. were used to obtain lyophilisates which were named as TML and TPL, respectively. The former clover taxon represents a little explored species, while the latter one is often found in dietary supplements and functional foods due to the content of isoflavones that alleviate various menopausal symptoms. Detailed phytochemical profiles of both lyophilisates were examined and compared using coupled chromatographic (RP-LC) and spectroscopic PDA/ESI-QToF/MS-MS methods. A total of 54 and 55 compounds were identified in TML and TPL, respectively. Close chemotaxonomic similarities were confirmed for both clover taxa examined, especially in terms of the most abundant isoflavones and hydroxycinnamates. For the first time, neurotropic polyphenols (AChE inhibitors) were identified in clover extracts using TLC-bioautography. In this group, caffeoylmalic acid exhibited significant AChE inhibitory effect confirming the potential of TPL and TML to support physiological functions of the endocrine and nervous systems.

The TLC-Bioautography as a Tool for Rapid Enzyme Inhibitors detection - A Review.

Microorganisms and plants can be important sources of many compounds with potential pharmaceutical applications. Extraction of these matrices is one of the ways of identifying the presence of inhibitory active substances against enzymes whose high activity leads to serious human diseases including cancer, Parkinson's or Crohn's diseases. The isolation and purification of inhibitors are time-consuming and expensive steps in the analysis of the crude extract and therefore, it is necessary to find a fast, efficient, and inexpensive method for screening extracts of interest. TLC-Bioautography combines the separation of the extract on a thin layer with its subsequent biological analysis. TLC-Bioautography methods have been developed for several classes of enzymes including oxidoreductases, hydrolases and isomerases, and there is a potential for developing functional methods for other classes of enzymes. This review summarizes known TLC-Bioautography methods and their applications for determining the presence of enzyme inhibitors in extracts and compares the effectiveness of different methodological approaches. It also indicates the current state and perspective of the development of TLC-Bioautography and its possible future applications.

Development of a validated HPTLC-bioautographic method for evaluation of aromatase inhibitory activity of plant extracts and their constituents.

INTRODUCTION: Aromatase is a CYP450 enzyme that catalyses the conversion of androgens into oestrogens, where the decrease in the production of oestrogens aided by aromatase inhibitors is considered a target in post-menopausal breast cancer therapy. TLC-bioautography is a technique employed for combining chromatographic separations on TLC plates with bioassays. This is the first report to evaluate aromatase inhibitory activity using this technique. OBJECTIVES: The aim of this study is to develop and validate a new TLC-bioautographic method for determination of aromatase inhibitory activity in 14 plant extracts. Two quantitation methods, the peak area method and reciprocal iso-inhibition volume (RIV) method, were compared and investigated to attain reliable results. Factors affecting the enzymatic reaction (temperature, pH, enzyme and substrate concentrations … etc.) were also investigated to attain the optimum parameters. METHODOLOGY: TLC assisted by digital image processing was implemented for quantitative estimation of the aromatase inhibition of 14 plant extracts using chrysin as positive control. The fluorometric substrate dibenzyl fluorescein (DBF) was utilised for the assay, where inhibitory compounds were visualised as dark spots against a blue fluorescent background. Two software programs, Sorbfil® videodensitometer (in the peak area method) and ImageJ® (in the RIV method), were thoroughly validated using the International Council on Harmonisation (ICH) guideline and used for quantitation. RESULTS: The RIV method showed superiority over the peak area method in the quantitation results of the tracks with non-homogenous background with %RSD values of 0.98 and 1.49 compared with 2.86 and 3.58, respectively. Further, the methods allow the comparison of the activity of different unknown inhibitory compounds without the need for a reference or a positive control. CONCLUSION: Using the TLC-bioautographic method by image processing combined with the RIV quantitation method, simultaneous separation and quantitation of aromatase inhibitory components could be applied to estimate the relative activity of various plant extracts.

Composition, Anti-MRSA Activity and Toxicity of Essential Oils from Cymbopogon Species.

Many of the essential oils obtained from medicinal plants possess proven antimicrobial activity and are suitable for medicinal purposes and applications in the food industry. The aim of the present work was the chemical analysis of 19 essential oils (EOs) from seven different Cymbopogon species (C. nardus, C. citratus, C winterianus, C. flexuosus, C. schoenanthus, C. martinii, C. giganteus). Five different chemotypes were established by GC/MS and TLC assay. The EOs, as well as some reference compounds, i.e., citronellol, geraniol and citral (neral + geranial), were also tested for their antimicrobial and antibiofilm activity against methicillin-resistant Staphylococcus aureus (MRSA) by the microdilution method and direct bioautography. The toxicity of EOs was evaluated by Danio rerio 'Zebrafish' model assay. All examined EOs showed moderate to high activity against MRSA, with the highest activity noted for C. flexuosus-lemongrass essential oil, both in microdilution and direct autobiography method. Significant difference in the toxicity of the examined EOs was also detected.

Fractionation of Lycopodiaceae Alkaloids and Evaluation of Their Anticholinesterase and Cytotoxic Activities.

In view of the abundant evidence that Lycopodiaceae alkaloids, including the well-known huperzine A (HupA), are among the potent acetylcholinesterase (AChE) inhibitors, an attempt was made to search for new compounds responsible for this property. For this purpose, three plant species belonging to the Lycopodiaceae family, commonly found in the Euro-Asia region, were subjected to the isolation of bioactive compounds, their identification and subsequent evaluation of their anticholinesterase and cytotoxic activities. Methanolic extracts of two Lycopodium and one Hupezia species were obtained via optimized pressurized liquid extraction (PLE) and then pre-purified using innovative gradient vacuum liquid chromatography (gVLC). For the first time, three sorbents of different porosity packed in polypropylene cartridges and mobile phase systems of different polarity were used to elute the target compounds. This technique proved to be a rapid tool for the obtainment of alkaloid fractions and allowed one to select the appropriate process conditions to yield potent AChE inhibitors in each of the species studied. More than 100 collected fractions were analyzed via HPLC/ESI-QTOF-MS, which enabled one to detect more than 50 compounds, including several new ones previously unreported. Some of them were present in high purity fractions (60-90% of the established purity). TLC bioautography assays proved that the analyzed species are rich sources of AChE inhibitors, but H. selago showed the highest anti-AChE activity. Additionally, the modified silanized silica gel sorbent used allowed one to isolate L. clavatum alkaloids more efficiently using an aqueous reversed-phase solvent system. Furthermore, the tested extracts from the three plant extracts were found to be safe, as they did not exhibit cytotoxicity to skin fibroblasts.

Development of Synergy-Based Combination for Learning and Memory Using in vitro, in vivo and TLC-MS-Bioautographic Studies.

The present study is aimed at developing a synergistic combination to enhance learning and memory in Alzheimer's patients with the help of eight common medicinal plants used in the AYUSH system. Aqueous and hydroalcoholic extracts of eight medicinal plants from the AYUSH system of medicine were prepared. These were subjected to in vitro anticholinesterase activity, to find out the combination index of synergistic combination. The synergistic combination and their individual extracts were subjected to total phenol, flavonoid and antioxidant activity estimation. Further, in vivo neurobehavioral studies in rats were carried out followed by TLC-MS-bioautographic identification of bioactive metabolites. Out of the sixteen extracts, aqueous extracts of Withania somnifera (L.) Dunal (WSA) and Myristica fragrans (L.) Dunal (MFA) were selected for the development of synergistic combination based on their IC50 value in vitro anticholinesterase assay. The synergistic combination inhibited the anticholinesterase activity significantly as compared to the individual extracts of WSA and MFA. The synergistic combination also showed more phenolic and flavonoid contents with potential antioxidant activity. The TLC-bioautography showed four white spots in WSA, signifying sitoindosides VII, VIII, quercetin, isopelletierine and Withanolide S as AChE inhibitory compounds while showing five white spots of anti-cholinesterase active metabolites identified as eugenol, methyl eugenol, myristic acid, galbacin and ß-sitosterol in MFA. The observation of neurocognitive behavior in amnesia induced subjects manifested that both the synergistic combinations showed comparable results to that of standard piracetam, though the synergistic combination containing a higher concentration of WSA showed more appreciable results in ameliorating dementia in rats. The study suggests that the synergy based combination successfully enhanced memory and learning by abating free radical and acetylcholine levels, and increased learning and memory in rats, providing a strong rationale for its use in the treatment of dementia and Alzheimer's disease.

Antimicrobial metabolite profiling of Nigrospora sphaerica from Adiantum philippense L.

BACKGROUND: Endophyte bestows beneficial aspects to its inhabiting host, along with a contribution to diverse structural attributes with biological potential. In this regard, antimicrobial profiling of fungal endophytes from medicinal plant Adiantum philippense revealed bioactive Nigrospora sphaerica from the leaf segment. Chemical and biological profiling through TLC-bioautography and hyphenated spectroscopic techniques confirmed the presence of phomalactone as an antimicrobial metabolite. RESULTS: The chemical investigation of the broth extract by bioassay-guided fractionation confirmed phomalactone as a bioactive antimicrobial secondary metabolite. The antimicrobial activity of phomalactone was found to be highest against Escherichia coli by disc diffusion assay. The MIC was found to be significant against both Escherichia coli and Xanthomonas campestris in the case of bacteria and dermatophyte Candida albicans at 150 µg/ml, respectively. CONCLUSIONS: Overall, the results highlighted the antimicrobial potential of phomalactone from the endophyte Nigrospora sphaerica exhibiting a broad spectrum of antimicrobial activity against human and phytopathogenic bacteria and fungi. This work is the first report regarding the antibacterial activity of phomalactone.

Acetylcholinesterase Inhibitors among Zingiber officinale Terpenes-Extraction Conditions and Thin Layer Chromatography-Based Bioautography Studies.

Although numerous studies have been conducted on ginger extracts and fractions, the data on the pharmacological activity of single constituents of Zingiber officinale are still insufficient. To assess the antidementia properties of the plant, a thin layer chromatography (TLC)-based bioautography acetylcholinesterase inhibitory assay was performed on the Zingiber officinale diethyl ether extract. It led to the recognition of three active inhibitors among volatile constituents of the plant: ar-curcumene (A), α-sesquiphellandrene (B) and a-zingiberene (C). The identification of the components was possible thanks to the application of a TLC-HPLC-MS interface analysis of active zones and the GC-MS qualitative analysis of the tested samples. Based on the obtained results, the influence of several extraction techniques (hydrodistillation-HD, pressurized liquid extraction or accelerated solvent extraction-ASE, shaking maceration-SM, supercritical fluid extraction-SFE, and ultrasound-assisted extraction-UAE) on the recovery of the active metabolites from plant material was assessed to deliver enriched extracts. As a result, HD and SFE, were found to be the most efficient methods to recover the volatile components and the concentrations of A, B, and C reached 0.51 ± 0.025, 0.77 ± 0.045, and 1.67 ± 0.11 percent, respectively. Only HD and SFE were found to recover monoterpene hydrocarbons from the plant matrix. The remaining techniques provided extracts rich in more complex constituents, like sesquiterpenes.

TLC-Bioautography as a fast and cheap screening method for the detection of α-chymotrypsin inhibitors in crude plant extracts.

TLC-Bioautography is a fast and effective method for assessing the inhibitory effect of compounds present in plant extracts against microbial species. However, this method has a hidden, currently underutilized potential for evaluating the presence of inhibitory compounds against selected enzymes. The aim of this work was to design a functional TLC-Bioautography method for the evaluation of protease inhibitors present in plant extracts. The method is based on the hydrolysis of Nα-benzoyl-DL-arginine-p-nitroanilide hydrochloride (BApNA) by α-chymotrypsin as a representative serine protease to produce coloured para-nitroaniline (pNA). Derivatization of pNA with both sodium nitrite and N-(1-naphthyl) ethylenediamine (NPED) leads to the formation of a pink azo dye. This step improves the resolution of active compounds on the chromatogram, which appear as light spots on a pink background. The developed method was tested for the analysis of protease inhibitors in different plant materials such as grape pomace from Vitis vinifera, Picea abies bark, Hippophae rhamnoides berries, Hordeum sativum bran, Triticum aestivum bran and Avena sativa bran. Plant extracts, which could not be analysed by a commonly used spectrophotometric method due to interference, were assessed by this method.

Synergistic Antibacterial Efficacy of Biogenic Synthesized Silver Nanoparticles using Ajuga bractosa with Standard Antibiotics: A Study Against Bacterial Pathogens.

BACKGROUND: Multi-drug resistance in bacterial pathogens is a major concern of today. Green synthesis technology is being used to cure infectious diseases. OBJECTIVES: The aim of the current research was to analyze the antibacterial, antioxidant, and phytochemical screening of green synthesized silver nanoparticles using Ajuga bracteosa. METHODS: Extract of A. bracteosa was prepared by maceration technique. Silver nanoparticles were synthesized using A. bracteosa extract and were confirmed by UV-Vis spectrophotometer, Scanning Electron Microscope (SEM) and Fourier Transform Infrared Spectroscopy (FTIR). The antibacterial, anti-biofilm, cell proliferation inhibition, TLC-Bioautography, TLC-Spot screening, antioxidant, and phytochemical screening were also investigated. RESULTS: UV-Vis spectrum and Scanning electron microscopy confirmed the synthesis of green nanoparticles at 400 nm with tube-like structures. FTIR spectrum showed that functional groups of nanoparticles have a role in capping and stability of AgNP. Agar well diffusion assay represented the maximum antibacterial effect of ABAgNPs against Escherichia coli, Klebsiella pneumoniae, Streptococcus pyogenes, Staphylococcus aureus, and Pseudomonas aeruginosa at 0.10 g/mL concentration compared to ABaqu. Two types of interactions among nanoparticles, aqueous extract, and antibiotics (Synergistic and additive) were recorded against tested pathogens. Crystal violet, MTT, TLC-bio-autography, and spot screening supported the findings of the antibacterial assay. Highest antioxidant potential effect in ABaqu was 14.62% (DPPH) and 13.64% (ABTS) while 4.85% (DPPH) and 4.86% (ABTS) was recorded in ABAgNPs. Presence of phytochemical constituents showed pharmacological importance. CONCLUSION: It was concluded that green synthesis is an innovative technology in which natural products are conjugated with metallic particles and are used against infectious pathogens. The current research showed the significant use of green nanoparticles against etiological agents.

Data on the inhibitory effect of endophytic fungi of traditional medicinal plants against pancreatic lipase (PL).

This article describes isolation and pancreatic lipase (PL) inhibitory potential of 18 endophytic fungi isolated from the various parts of six indigenous medicinal plants. PL catalyzes absorption and hydrolysis of triglycerides into di-glycerides into mono-glycerides and free fatty acids. PL inhibitors are well-known for the disruption of pancreatic lipase activity. The quest for novel pancreatic lipase inhibitors is crucially important owing to their therapeutic potential in the treatment of obesity and related chronic diseases. The present dataset provides information about the presence of endophytic fungi in the internal tissues of selected plants and the PL inhibitory potential of their metabolites using bioassay based screening. Absence of the yellow zone surrounding the standard Orlistat and test extract indicated PL inhibition due to the cumulative effect of metabolites present in the extract. The data suggests that TLC bio-autographic method is simple, rapid and reproducible and therefore it could be effectively used for high throughput screening of PL inhibitors from natural sources.

Molecular profiling of endophytic Streptomyces cavourensis MH16 inhabiting Millingtonia hortensis Linn. and influence of different culture media on biosynthesis of antimicrobial metabolites.

Endophytic actinomycetes, a prolific source of natural products, are well known for their diverse metabolic versatility, and their association with medicinal plants and antimicrobial potential are well worth exploring. We isolated and identified the Streptomyces cavourensis strain MH16 inhabiting the tree Millingtonia hortensis Linn. using phylogenetic analysis based on a 16S rRNA molecular approach. We used the disc diffusion method to evaluate the impact of differences in the compositions of the media on the production of secondary metabolites from strain MH16. The production of antimicrobial metabolites was determined by the observation of inhibition zones on intensive bands when using a TLC-bioautography assay. Biosynthesis of secondary metabolites was optimal when the strain MH16 was cultured in ISP-2 medium as depicted by a zone of inhibition. Strain MH16 effectively inhibited methicillin-resistant Staphylococcus aureus, Escherichia coli, Candida albicans, and other multi drug-resistant pathogens. The minimum inhibitory concentration of the antimicrobial metabolites was 25-100 µg mL-1. The study manifests the optimization and utilization of different fermentation media which best suits for increased production of the secondary metabolites from Streptomyces cavourensis. This research suggests that the antimicrobial metabolites of strain MH16 found in M. hortensis has great potential for the biodiscovery of new anti-infective drugs against a wide range of multidrug-resistant pathogens.

Antimicrobial Potential of Single Metabolites of Curcuma longa Assessed in the Total Extract by Thin-Layer Chromatography-Based Bioautography and Image Analysis.

Curcuma longa from Zingiberaceae belongs to the major spices consumed around the world, known from its cholagogue, anti-inflammatory, and antimicrobial properties. Lack of data on the activity of single components of turmeric extract encouraged the authors to apply TLC (thin-layer chromatography) based bioautography studies to reveal its antimicrobial constituents and construct a universal platform for the bioactivity assessment of crude extracts, with help of a freeware ImageJ software. This optimized chromatographic bioassay performed on diethyl ether and methanol extracts of Curcuma longa was successfully applied on the total extract and revealed the antimicrobial potential of single components against a variety of Gram-positive strains, with no need for their isolation from the mixture. The obtained results were further confronted with a classic microdilution antimicrobial assay on the isolates, purified from the crude extracts by centrifugal partition chromatography in the following solvent system: heptane-chloroform-methanol-water (5:6:3:2) (v/v/v/v).

Anti-inflammatory and antioxidative effects of the buds from different species of Populus in human gingival fibroblast cells: Role of bioflavanones.

BACKGROUND: Poplar leaf-buds (Populi gemmae) are used traditionally as anti-inflammatory agents to the treatment of skin injuries or cough. They differ in their diverse chemical composition and different types of activities, whose mechanisms are not fully recognized. PURPOSE: Evaluation and comparison of anti-inflammatory activity of leaf-buds extracts from Populus nigra, P. × berolinensis and P. lasiocarpa and flavanones - pinocembrin and pinostrobin towards human gingival fibroblasts (HGF-1) pro-inflammatory stimulated by silver nanoparticles (AgNPs). Determination of antioxidant activity associated with anti-inflammatory properties by means of bioautographic TLC tests. METHODS: Phytochemical analysis was performed by TLC and videodensitometry analysis. The extracts were standardized on the pinocembrin and pinostrobin content. Bioautography was performed using 2,2-diphenyl-1-picrylhydrazyl (DPPH) and riboflavin-light blue tetrazolium chloride (riboflavin-light-NBT) radicals to assess the extracts and both flavanones radical scavenging properties as well as potential inhibition of xanthine oxidase (XO) activity. The protective effects of poplar buds extract and flavanones - pinocembrin and pinostrobin on HGF-1 line exposured to AgNPs were investigated by analysis of interleukin 6 (IL-6) and interleukin IL-1ß (IL-1ß) level measured by ELISA kit. The messenger ribonucleic acid (mRNA) of both cytokines was determined by real-time quantitative PCR. The involvement of cyclooxygenase 2 protein (COX-2) was studied using Western blot analysis. RESULTS: The presence of several flavanones and phenolic acids, which have radical scavenging properties, was revealed in all of the bud poplar extracts analyzed. Treatment with particular flavanones or extracts from buds of P. × berolinensis and P. nigra decreased the IL-6 and IL-1ß release in HGF-1 cells and down-regulation of mRNA for both cytokines was observed. The COX-2 protein expression was demonstrated for pinocembrin and P. × berolinensis buds. These effects were not observed for buds from P. lasiocarpa not containing of flavonoids. CONCLUSION: The potential protective role of pinocembrin and pinostrobin and extracts from buds P. nigra and P. × berolinensis against AgNPs induced inflammation and cytotoxicity in HGF-1 cells is disclosed. In addition, the antioxidant properties of poplar bud extracts have been demonstrated. P. × berolinensis buds showed the highest activity in both the in vitro model and in the bioautographic tests.

TLC-bioautography directed isolation of antibacterial compounds from active fractionation of Ferula ferulioides.

A novel optimised isolation method, TLC-bioautography, was evaluated and utilised in this research. Antibacterial compounds which were isolated from the dichloromethane extract of Ferula ferulioides (Steud.) Korovin were detected by means of the method. Their structures were elucidated by extensive spectral and chemical methods. Their antibacterial activities against drug-resistant Staphylococcus aureus (S. aureus) strains were evaluated with broth microdilution method, and the results proved that TLC-bioautography was an effective and highly efficient way to screen natural compounds from plant extracts against drug-resistant strains.

Phytochemical analysis and mode of action against Candida glabrata of Paeonia emodi extracts.

In the present study, we have evaluated the antifungal activity of the seed, root and leaf of Paeonia emodi (commonly known as Himalayan peony) in four common solvents (acetone, chloroform, methanol and water) against six fungal strains. The methanolic seed extract (MSE) showed promising antifungal activity against Candida albicans (6.25mg/mL), Candida glabrata (3.12mg/mL) and Candida parapsilosis (12.50mg/mL) among all the fungal strains tested. Combination of the MSE with the well-known commercial antifungal drugs amphotericin B (Amp B), nystatin (NYS) and fluconazole (FLC) resulted in the killing of C. glabrata at non-inhibitory concentrations, i.e., 0.35µg/mL for Amp B, 0.55µg/mL for NYS and 1.19µg/mL for FLC. Notably, MSE caused cell wall damage of C. glabrata cells, as confirmed by confocal microscopy, flowcytometry and scanning electron microscopy (SEM). The MSE was fractionated by thin layer chromatography (TLC). TLC-bioautography was used to determine the active compounds present in the MSE. Column chromatography was used to separate the potential active compounds from the MSE. Furthermore, gas chromatography-mass spectrometry (GC-MS) andfourier-transform infrared spectroscopy (FTIR) were used to identify the phytocomponents of the MSE. These experiments revealed 13-docosenamide/9-octadecenamide/trans-13-docosenamide (89.70%) as being the predominant compound using a chloroform/methanol solvent system for the separation. Interestingly, the MSE also exhibited less significant cytotoxicity at the minimum inhibitory concentration (MIC) against mammalian cells (HeLa and HEK293). This study suggests that the MSE of P. emodi can be used for the treatment of C. glabrata infection.

Biological activities, and phytocompounds of northwest Algeria Ajuga iva (L) extracts: Partial identification of the antibacterial fraction.

The use of synthetic food additive and the appearance of antibiotic resistance are at the basis of important human health problems. The substitution of synthetic compounds with new natural substances extracted from plants or microorganisms is therefore the ideal solution to this scourge. The objective of this work was to evaluate the phyto-constituents (polyphenols, flavonoids and condensed tannins), and to test the biological activities (antioxidant, antibacterial and antiviral) of the Ajuga iva (L) aerial part extracts. The antioxidant activity assayed by DPPH method showed an IC50 of 0.43 ±â€¯0.03 mg/mL. Antibacterial activity of aqueous and hydro methalonic extracts was tested against seven pathogenic bacteria (Staphylococcus aureus, Methicillin Resistant Staphylococcus aureus (MRS), Listeria monocytogenes, Pseudomonas aeruginosa, Bacillus cereus, Escherichia coli and Salmonella enteritidis) using the diffusion method. A Thin Layer Chromatography-bioautotography-guided was performed, and the isolated antibacterial fraction was identified by CG-MS analysis. Antiviral effect of methanolic extract performed on 4 viruses: Coxsackie Virus type B-3 (CVB-3), Adenovirus type 5 (ADV-5), Respiratory Syncytial Virus type B (RSV-B) and Herpes Simplex Virus type 2 (HSV-2) showed an activity against Coxsackie Virus. As a result of this study, the aerial parts of Ajuga iva (L) extract could be used in the food, cosmetic, medical and health sectors.

Improved bioautographic assay on TLC layers for qualitative and quantitative estimation of xanthine oxidase inhibitors and superoxide scavengers.

A new agar-free bioautographic assay for xanthine oxidase (XO) inhibitors and superoxide scavengers on TLC layers was developed and validated. Compared to the first version of TLC bioautographic agar overlay method, our bioautographic assay greatly improved the sensitivity and quantification ability. The limit of detection (LOD) of this assay was 0.017ng for allopurinol. Quantitative estimation of XO inhibitors and superoxide scavengers was achieved by densitometry scanning, expressed as allopurinol equivalents in millimoles on a per sample weight basis. This assay has acceptable accuracy (95.37-99.23%), intra-day and inter-day precisions (RSD, 2.56-6.69%), as well as intra-plate and inter-plate precisions (RSD, 2.93-9.62%). Six pure compounds and three herbal extracts were evaluated for their potential XO inhibitory and superoxide scavenging activity by this bioautographic assay on TLC layers. Four active components were separated, located and identified in Astragalus membranaceus var. mongholicus extract by the bioautographic assay after TLC separation. The developed method is rapid, simple, sensitive and stable for screening and estimation of the potential XO inhibitors and superoxide scavengers.