Through Thin Layer Chromatography Research Articles

Characterization of antibiofilm compound from marine sponge Stylissa carteri.

The fouling phenomenon grabbed global attention and caused huge economic losses specifically in marine-related industries. Sessile behavior exposed the sponge to the risk of fouling. However, their bodies remained free from foulers, which were attributed to the chemical defense system. The objectives of this study were to determine the antibiofilm activity of the marine sponge, Stylissa carteri, and to characterize the isolated compound involved. The antibiofilm activity of S. carteri methanolic crude extract (MCE) and fractions was tested against biofilm-producing bacteria, Pseudomonas aeruginosa, using two different modes of crystal violet biofilm assays: preventive and detachment. Besides that, the disc-diffusion test was conducted to screen the antibacterial activity against gram-positive and gram-negative bacteria while a cytotoxicity assay was conducted on the HepG2 cell line. Bioassay-guided fractionation was carried out using vacuum liquid chromatography (VLC) and solid phase extraction using a C18 Sep-Pak Cartridge. The crystal compound was isolated and characterized through thin-layer chromatography (TLC), Fourier transform infrared (FTIR) spectroscopy, liquid chromatography-mass spectrometry (LCMS), and nuclear magnetic resonance (NMR) spectroscopy. The S. carteri MCE showed a promising result with a half-maximal inhibitory concentration (IC50) of 20.22μg/mL in the preventive assay, while no IC50 was determined in the detachment assay since all inhibitions < 50%. The S. carteri MCE exhibited broad-spectrum antibacterial activity and displayed a non-cytotoxic effect. Fraction 4 from MCE of S. carteri (IC50 = 2.40μg/mL) reduced the biofilm in the preventive assay at all concentrations and exhibited no antibacterial activity indicating the independence of antibiofilm from antibacterial properties. Based on the data obtained, an alkaloid named debromohymenialdisine (DBH) was identified from Fraction 4 of S. carteri MCE. In conclusion, S. carteri was able to reduce the establishment of the biofilm formed by P. aeruginosa and could serve as a prominent source of natural antifouling agents.

Neurospora sp. Mediated Synthesis of Naringenin for the Production of Bioactive Nanomaterials.

The application of Neurospora sp., a fungus that commonly thrives on complex agricultural and plant wastes, has proven successful in utilizing citrus peel waste as a source of naringin. A UV-Vis spectrophotometric method proved the biotransformation of naringin, with an absorption maximum (λmax) observed at 310 nm for the biotransformed product, naringenin (NAR). Further verification of the conversion of naringin was provided through thin layer chromatography (TLC). The Neurospora crassa mediated biotransformation of naringin to NAR was utilized for the rapid (within 5 min) synthesis of silver (Ag) and gold (Au) nanoconjugates using sunlight to accelerate the reaction. The synthesized NAR-nano Ag and NAR-nano Au conjugates exhibited monodispersed spherical and spherical as well as polygonal shaped particles, respectively. Both of the nanoconjugates showed average particle sizes of less than 90 nm from TEM analysis. The NAR-Ag and NAR-Au nanoconjugates displayed potential enhancement of the antimicrobial activities, including antibacterial and nematicidal properties over either standalone NAR or Ag or Au NPs. This study reveals the potential of naringinase-producing Neurospora sp. for transforming naringin into NAR. Additionally, the resulting NAR-Ag and NAR-Au nanoconjugates showed promise as sustainable antibiotics and biochemical nematicides.

Assessment of total aflatoxin and ochratoxin A in poultry feed ingredients by thin-layer chromatography and enzyme-linked immunosorbent assay.

This study was conducted to evaluate the prevalence of total aflatoxin (AF) and ochratoxin A (OTA) in poultry feed ingredients under different environmental conditions during the summer and winter seasons, while the hygiene quality of the feed ingredient was assessed through viable fungal count (VFC). A total of 288 poultry feed ingredients (n = 96 each) samples were collected from different poultry shops, which were initially analyzed for the presence of AF and OTA through thin layer chromatography (TLC) and then confirmed the contamination concentration through the enzyme-linked immunosorbent assay method. The results of the current study confirmed the incidence of contamination with AF and OTA by TLC and ELISA methods. The contamination level of AF ranged from 26.09 to 50.56 (mean = 41.22 ± 9.45) μg/kg, whereas the contamination level of OTA ranged from 50.13 to 6.21 (mean 42.60 ± 6.21) μg/kg. The contamination level of AF was found to be above the permissible level set by the Food and Drug Administration (20 μg/kg), whereas the contamination level of OTA was below the permissible limits. Moreover, the VFC values were also below the recommended level. The results showed that the association between AF, OTA, and moisture content was significant (p < 0.05). Mycotoxin contamination was significantly (p < 0.05) highest in the winter season. These findings suggested that continuous monitoring regimes might prevent mycotoxin contamination in poultry feed ingredients.

Bis-indole based triazine derivatives: Synthesis, characterization, in vitro β-glucuronidase anti-cancer and anti-bacterial evaluation along with in silico molecular docking and ADME analysis

The present work described the synthetic procedure adopted for the synthesis of bis-indole-based triazine derivatives via a series of reactions. All the compounds were synthesized through a stepwise reaction. After being confirmation through thin-layer chromatography (TLC) these compounds were characterized through various spectroscopic techniques including 1HNMR, 13CNMR and HREI-MS and evaluated against beta-glucuronidase in the presence of standard drug D-saccharic acid 1,4 lactone (IC50 = 31.2 ± 1.0 µM). Most of the synthesized derivatives were found with better to good inhibitory potentials (IC50 = 5.30 ± 2.0 µM to 33.10 ± 1.0 µM). SAR explains better results of analogs due to the presence of varied substituents on the aromatic rings. In this regard, the excellent potential was shown by analogs 1, 3, 6, 8 and 9 with IC50 = 5.30 ± 2.0, 7.10 ± 4.0, 6.10 ± 3.0, 8.40 ± 1.0 and 7.20 ± 3.0 µM respectively). In addition, all the synthesized analogs were evaluated for anti-cancer and anti-bacterial (E. coli) activities in which the targeted compounds were found with significant potentials in the presence of standard drugs Tetrandrineb (IC50 = 1.37 ± 0.10 µM) and streptomycin respectively. These derivatives were further subjected to molecular docking studies in order to investigate better binding sites with distance. Additionally, ADME properties for the synthesized compounds were also explored the drug like properties of the synthesized compounds.

Selection and identification of antibacterial and GABA-producing lactic acid bacteria from Vietnamese fermented meat

This study aimed to screen the lactic acid bacteria (LAB) strain that has high potential in gamma-aminobutyric acid (GABA) production and antibacterial properties, which could be applied in Vietnamese fermented meat (Nem chua) fermentation. These bacterial strains were isolated from commercial Nem chua products and stored in the laboratory. GABA synthesized from 18 strains of LAB in the MRS media adding 3% MSG was qualitative and quantitative analyzed through thin layer chromatography (TLC) and its calibration curve equation, then antibacterial activity and lactic acid production were also tested by acidic titration method, and well diffusion method, respectively. The results showed that all LAB strains synthesized GABA with a concentration from 1.323 - 1.582 mg/mL; however, eight strains were recorded with higher potential, including VL1, XP1, TP1, HC2, CP2, LV2, TX7, and HK 162. These eight strains showed the inhibition zone to Escherichia coli indicator from 4.67 - 11.67 mm. Besides, they are capable of yielding lactic acid from 18.00 - 19.35 g/L after seven days of fermentation in MRS media. The strain VL1 with the highest potential was identified as Lactobacillus plantarum. This strain was suggested as a promising starter to be applied in the Nem chua fermentation process.

Molecular Characterization of Ochratoxin a Producing Indigenous Aspergillus Strains from Poultry Feed in Pakistan

Ochratoxin A (OTA) is nephrocarcinogenic and immunosuppressive toxin and OTA producing molds contaminate the food crops. Isolation and identification of ochratoxin producing fungi was carried out from poultry feed samples (n=120) followed by preliminary confirmation through macroscopic and microscopic characteristics. Purified fungal isolates identified as Aspergillus 1842(91.68%) followed by Penicillium 91 (4.53%), Mucor 52 (2.58), Alternaria 7 (0.35%), Cladosporium 6 (0.29%), Fusarium 4 (0.199%) and un-identified (07). OTA production was confirmed through thin layer chromatography (TLC) followed by high performance liquid chromatography (HPLC). Only 41 isolates (2.22%) out of 1842 Aspergillus isolates were able to produce toxin. At genetic level, characterization was performed through polymerase chain reaction (PCR) using species specific gene primers. From 41 isolates 27, 9 and 5 were characterized as Aspergillus terreus, Aspergillus parasiticus, and Aspergillus ochraceus respectively. Physical and chemical factors were optimized for OTA production. Under the effect of 37 °C temperature and 7.5 pH of Sabouraud dextrose broth (SDB) medium, higher toxin (969.45±.03 µg/mL) production was observed from ASPO-6 isolate. ASPO-4 isolate produce higher toxin amount in SDB medium with supplementation of maize 5%, wheat 1% and rice 3%. OTA stability was determined by adjusting standard concentration of 100 µg/mL in organic solvents (chloroform, acetonitrile and methanol) and organic solids. Least percentage log reduction in OTA concentration and stability of OTA was observed in opaque vials with chloroform and sucrose and transparent vials with sucrose after 6 months. OTA can be used as indigenous standard for identification of OTA from field samples.

Benzothiazole—An Antifungal Compound Derived from Medicinal Mushroom Ganoderma lucidum against Mango Anthracnose Pathogen Colletotrichum gloeosporioides (Penz and (Sacc.))

The present investigation is focused on exploring the possibilities of identifying biomolecules from the fruiting body of the medicinal mushroom Ganoderma lucidum against the mango anthracnose pathogen Colletotrichum gloeosporioides. The fruiting body (cap and stipe portion) of G. lucidum extracted with ethyl acetate solvent at a maximum inhibitory concentration of 1 percent exhibited the maximum mycelial growth inhibition of C. gloeosporioides with 70.10 percent and 40.77 percent, respectively. Furthermore, subjecting the ethyl acetate extracts from the cap portion of G. lucidum through thin layer chromatography (TLC) revealed the presence of two bands with Rf values of 0.38 and 0.35. The compounds eluted from band 1 recorded with the maximum mycelial growth inhibition of C. gloeosporioides by 53.77 percent followed by band 2 (46.33 percent) using an agar well diffusion test. Similarly, the analysis of ethyl acetate extracts from the cap portion of G. lucidum through Gas Chromatography-Mass spectroscopy (GC-MS) revealed the presence of the organoheterocyclic compound benzothiazole, as expressed in the highest peak area at 22.03 RT with the highest probability percentage (97%). Confirmation of the antifungal nature of benzothiazole was obtained by testing the standard sample of benzothiazole which showed a cent percent of inhibition on mycelial growth of C. gloeosporioides at 50 ppm minimum fungicidal concentration. Furthermore, benzothiazole caused abnormality in the mycelial structures, viz., distortion, shrinkage, clumping of mycelium, conidial malformation, and complete arrestment of conidial germination of C. gloeosporioides as observed through Scanning Electron Microscopy. The research on biomolecular extract of G. lucidum could be a novel and interesting concept for the possibility in suppression of plant pathogenic microbes in the natural field.

Molecular targeting of prodigiosin against anti-inflammatory genes cyclooxygenase-1 and -2

The prodigiosin produced by the bacteria Serratia marcescens (SM) has been isolated and optimized with the streak plate method from different natural sources, such as human urine (SM1), soil (SM2), vegetable matter (Cucurbita maxima, SM3) and water (SM4). The isolated prodigiosin was confirmed as SM by its morphological characteristics, genomic size (1.5 kb), identification, and protein purification (4 kDa) using SDS-PAGE. Further, the total cell protein was confirmed and purified through thin layer chromatography (TLC) and high performance liquid chromatography (HPLC). The prodigiosin exhibited good microbial activity against selected Gram-positive bacteria and fungi. The binding efficiency of prodigiosin against cyclooxygenase-1 (COX-1) and cyclooxygenase-2 (COX-2) genes was assessed using an in silico molecular docking method that suggested higher binding efficiency for COX-2 than COX-1. Further, prodigiosin was nontoxic to rat islet (RIN-5F) cells in an assay with 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenylteterazolium bromide) in vitro, and its morphological characteristics of cell shape, clumping, chromatin, and nuclear morphology were normal when viewed under bright light microscopy. Finally, prodigiosin was tested for anti-inflammatory properties against lipopolysaccharide-induced RIN-5F cells. The tests revealed that prodigiosin protected against elution of inflammatory markers like NO, interleukin-6 (IL6), and IL8 in a dose-dependent manner.

Bioconversion of lovastatin to simvastatin by Streptomyces carpaticus toward the inhibition of HMG-CoA activity.

The aim of this study was the modification of lovastatin by microbes to improve its potential. Actinobacteria exhibit staggering diversity in terms of their biosynthetic capability for specialized metabolites which has been traced back to the presence of specialized gene clusters. The objective of the study is to exploit the potential of Actinobacteria strain(s), which can biotransform lovastatin to simvastatin, which might be a more potent therapeutic agent than lovastatin. We have screened 40 Actinobacteria strains and assessed their biotransformation potential primarily through thin layer chromatography (TLC) analysis, followed by high performance thin layer chromatography and high performance liquid chromatography analysis. One strain C7 (CTL S12) has been identified as a potential Actinobacteria that favored the simvastatin biotransformation. The morphological and biochemical analysis together with 16S rRNA sequencing coupled with phylogenetic analysis confirmed the ideal strain (C7) as Streptomyces carpaticus. Successively, the purified simvastatin from S. carpaticus was characterized by liquid chromatography-mass spectrometry (LC-MS), infrared spectrometry, nuclear magnetic resonance, and HMG-CoA assay. In the LC-MS analysis, a peak at 419.24 m/z confirmed the elemental composition of simvastatin (C25 H39 O5 ). In HMG-CoA assay, the IC50 of simvastatin was 50μg/ml, and the inhibitory potential was 1.36 times higher compared to that of lovastatin. Thus, the biotransformation of simvastatin from lovastatin by S. carpaticus is reported for the first time.

2-Mercaptobenzimidazole Based Hydrazone Derivatives as Potential Antioxidant and α-Glucosidase Inhibitors

Background: Schiff bases are organic compounds and play a vital role in making biologically active compounds in various fields of chemistry. It shows antioxidant, antidepressant, antimalarial, anti-inflammatory, antiglycation, and antimicrobial activity. Objective: Our current study is focused on synthesizing thirty-four 2-mercaptobenzimidazole (MBI) based novel hydrazone derivatives (09-42) which were examined for antioxidant free radical scavenging activity via both 1,1-diphenyl-2-picrylhydrazyl (DPPH) and hydrogen peroxide (H2O2) assay and explored their alpha-glucosidase inhibitory potential at various concentrations. Methods: Multistep reactions were involved in the synthesis of 2-mercaptobenzimidazole-based hydrazone derivatives. All steps of the reaction were carried out under different conditions through a reflux condenser to get the final target products, and the reaction was monitored regularly in each step through thin layer chromatography (TLC). The antioxidant and α-glucosidase inhibition assay was performed. Results:: Obtained results of antioxidants confirmed that compounds 42 (IC50 = 27.21μg/mL), 36 (IC50 = 27.90μg/mL), 23 (IC50 = 28.10μg/mL) and 35 (IC50 = 45.60μg/mL) possess excellent potential activity compared to standard ascorbic acid having (IC50 = 60.15 μg/mL) in DPPH assay. While in the case of H2O2 three compounds 38 (IC50 = 51.45 μg/mL), 15 (IC50 = 53.50 μg/mL), and 42 (IC50 = 60.42 μg/mL) showed excellent activity as compared to standard Gallic acid having (IC50= 60.67 μg/mL). In the screened compounds against alpha-glucosidase, compound 14 (IC50 = 162 μg/mL) was found to be the most active in the whole series. Another active compound 42 (IC50 = 237μg/mL) possessed moderate inhibitory potency against α-glucosidase enzyme. Conclusion: The different biological activities of these novel compounds may be due to different groups in the main skeleton of 2-mercaptobenzimidazole. Further experimental analysis and assessment of these compounds are important because they may lead to better antioxidants used in foods, cosmetics, and health-related products and act as antidiabetic drug development.

Larvicidal, antioxidant and biotoxicity assessment of (2-(((2-ethyl-2 methylhexyl)oxy)carbonyl)benzoic acid isolated from Bacillus pumilus against Aedes aegypti, Anopheles stephensi and Culex quinquefasciatus.

Mosquitoes are a vector for many dreadful diseases known for their public health concern. The continued use of synthetic insecticides against vector control has led to serious environmental impacts, human health problems, and the development of insect resistance. Hence, alternative mosquito control methods are needed to protect the environment and human health. In the present study, the bioefficacy of (2-(((2-ethyl-2 methylhexyl)oxy)carbonyl) benzoic acid isolated from Bacillus pumilus were tested against Aedes aegypti, Culex quinquefasciatus and Anopheles stephensi. The isolated bioactive compound was characterized through thin layer chromatography (TLC), UV-visible spectroscopy (UV), Fourier-transform infrared spectroscopy, nuclear magnetic resonance spectroscopy, and gas chromatography-mass spectrometry analysis. The pure compound caused a high percent mortality rate in a dose-dependent manner, the obtained values were 96, 82, 69, 50 and 34%; 86, 72, 56, 43, and 44%; 100, 90, 83, 70 and 56% against Ae. aegypti, Cx. quinquefasciatus, and An. stephensi respectively. The effective lethal concentration values (LC50) were 13.65, 14.90 and 9.64ppm against Ae. aegypti, Cx. quinquefasciatus, An. Stephensi, respectively. The effect of (2-(((2-ethyl-2 methylhexyl)oxy)carbonyl) benzoic acid significantly increased the superoxide dismutase, catalase, α, β esterase and Glutathione-S-transferase level after 24h of the treatment period. The comet assay confirmed that isolated compound causes DNA damage in all tested insects. Histopathological examinations of treated larvae showed shrunken body posture, damaged epithelial cells and microvillus as compared to control organisms. The biosafety of the isolated compound was assessed against G. affinis and did not produce mortality which confirmed that the activity of the isolated compound is species specific. The current study concludes that the critical success factors of new insecticidal agent development are based on the eco-compatibility and alternative tools for the pesticide producing industry.

Analysis of pharmacognostical standardization, antioxidant capacity and separation of phytocompounds from five different vegetable peels using different solvents

Vegetables are one of the most preferred food commodities and can be consumed either raw or as processed due to their health-promoting nutrients. In the present work, analysis of pharmacognostical standards, antioxidant capacity, and separation of phytocompounds through thin layer chromatography (TLC) from cabbage, cauliflower, pea, carrot, and potato peels were carried out. Microscopic analysis revealed the presence of wood fibers, trichomes, crystals, and annular xylem vessels in the vegetable peels. Physicochemical analysis showed that all the vegetable peel samples which were analysed have low (7.08%-10%) moisture content. The total ash content of vegetable peels varied in cauliflower peels (1.95±0.58) to the peels of pea (19.86±1.9). The content of acid insoluble ash varied from 1.46±0.63 to 3.09±0.59 in cauliflower and pea. Potato peel has the lowest water-soluble ash content (1.16±1.90) as compared to other peels. The highest pH value was found in the peels of pea (7), while the lowest pH was found in the peels of cabbage (4). Among all extracts, the petroleum ether extract has shown the greatest yield (5.6±0.45). The fluorescence analysis showed various colours like green, brown, pale green, and yellow under different chemical treatments. Different types of pri-secondary metabolites were detected in small, moderate, and high amounts and notified to provide numerous health benefits to humans. In case of DPPH assay, aqueous extract of cauliflower has shown the low value of IC50 (24.82 µg/ml) in comparison to standard, suggested the higher antioxidant activity of the extract. Among all the extracts, aqueous and methanol extracts of cauliflower have shown the better reducing and total antioxidant activity in comparison to standard. TLC profiling of methanolic extract of cabbage and cauliflower peels revealed the presence of different compounds of varying Rf values. Above results indicate that the food waste consists of valuable components and may be utilized as noticeable and cheap source in pharmaceuticals for the treatment of several life-threatening diseases.

Larvicidal and pupicidal activity of crude ethyl acetate extract fraction-7a of Cymodocea serrulata on Culex quinquefasciatus

Finding a potential bioactive compound against a specific mosquito vector is an urgent requirement. Thus, this research was designed to separate the significant compounds present in the crude ethyl acetate extract of Cymodocea serrulata through Thin Layer Chromatography (TLC), Column Chromatography (CC), and High Performance Liquid Chromatography (HPLC). Furthermore, the larvicidal and pupicidal activity of different concentrations (100, 300, 500, 700, and 900 g mL−1) of separated fraction against 4th instar larvae of Culex quinquefasciatus in 24 h of treatment. Interestingly, 7 fractions were obtained from crude ethyl acetate extract by TLC. One fraction (7a) from out of 7 was further separated into fractions 7a and 7b through CC and preparative TLC. Furthermore, the HPLC analysis result revealed that the fraction-7a contained 2 major peaks and 25 minor peaks. The fraction 7a was selected based on the Rf value and examined their larvicidal and pupicidal activity on 4th instar larvae and pupa of C. quinquefasciatus. Interestingly, the increased concentration (900 µg mL−1) of fraction-7a of ethyl acetate extracts of C. serrulata showed excellent larvicidal and pupicidal activities as 98.66 ± 1.88 % and 98 ± 5.65 %, respectively. These findings imply that the fraction 7a of C. serrulata ethyl acetate extract could be used as a mosquito vector control agent.

Lipophilic Constituents of the Blubber from Blue Whale, Balaenoptera musculus, Washed Ashore at Pakistan Coast

The blue whale, Balaenoptera musculus (Linnaeus, 1758), is the biggest animal recognized to exist today throughout world’s oceans. High commercial value of lipids has made this species vulnerable. Blubber, a crucial adaptation for mammals living in water, serves as energy reservoir. Surplus energy is deposited in the form of fatty acids (FA) and therefore have been analysed. The compositional analysis also helps in understanding the dietary and structural role of FAs in blubber. Lipid analysis of blubber from stranded, dead blue whale through Thin layer chromatography (TLC) has resulted in identifying 6 constituents. These constituents are a triacyl glyceride (TAG), 2 steroids, and 3 FAs. Approximate analysis of waxy constituents has also been attempted exploiting TLC. Gas chromatography-mass spectrometry (GC-MS) analyses has resulted in identification of 86 compounds, which were further confirmed through the Retention Indices. Altogether 17 SFAs including 4 Branched FAs, 5 MUFAs, and a PUFA were identified. These accounted to a total concentration of 85.7, 86.1, 84.8, and 89.7 % in jaw, abdomen, peduncle, and fluke, respectively. The main reasons for the low quantitative and qualitative content of PUFAs were susceptibility of PUFAs towards oxidation. Thus 8 FAlds, 4 FAlcs, and 3 other oxygenated FAs, which made a total of 2.7, 0.9, 1.3, and 5.2% in jaw, abdomen, peduncle, and fluke, respectively were justified. Further the chromatographic region where PUFAs are expected to resolve has been found masked with significant concentration of anthropogenic compounds, which accounted to 43.4, 35.6, 34.6, and 30.7% in jaw, abdomen, peduncle, and fluke, respectively. These pollutants included 25 hydrocarbons, 4 phthalates, 2 siloxanes, 2 bisphenols, and diphenyl carbonate. 4 natural Prenols were also identified. Altogether 16 constituents with concentration of 14.2, 8.0, 15.7, and 10.8% in jaw, abdomen, peduncle, and fluke, respectively, were remained unidentified. Few constituents were justified through food chain.

Extraction and Purification of Secondary Metabolites of Paris polyphylla (Smith) Using Column Chromatography.

Paris polyphylla Smith is an erect and herbaceous plant. It has rich chemical constituents such steroidal saponins, phytosterols, flavonoids, and alkaloids that possess antimicrobial and anticancer activities. In the current investigation, we examined the effect of different solvents in the extraction yield and further purification via column chromatography. The ground powdered rhizomes of P. polyphylla was extracted with 100% hexane, 100% ethyl acetate, 70% ethanol, and 70% methanol. The extracts were filtered, evaporated using a rotary evaporator, concentrated, and measured. Subsequently, the solvent with high extraction yield was further purified using column chromatography. Ethanol produced the highest extraction yield as compared to the other solvents. About 30 fractions were eluted which was pooled into four fractions based on the Rf values and bands observed through thin layer chromatography (TLC) analysis.

Removal of aflatoxin M1 from milk and aqueous medium by indigenously isolated strains of W. confusa H1 and L. plantarum S2

Employing LAB and/or their metabolites to reduce aflatoxins from food products is one of the fascinating areas of research. Therefore, in the present work an attempt was made to study the effect of isolated lactic acid bacteria (LAB) on the reduction of aflatoxin M1 (AFM1) from phosphate buffer saline (PBS) and reconstituted skim milk (RSM). Out of 68 isolates, nine strains of LAB isolated from food products were examined to bind and reduce AFM1. The isolates were incubated with AFM1 at different time and temperature (24 h, 48 h, and 72 h at 37 °C and 4 °C) and later the residual toxin in the supernatant was determined. The preliminary results through thin layer chromatography (TLC) revealed that all isolates were able to bind AFM1 in PBS. However, based on the intensity of spots observed in TLC sheets, it was indicated that two isolates, H1 and S2 genotypically identified as W. confusa and L. plantarum shown better AFM1 binding ability as compared to other isolates. Further, the heat treated cells of both the selected strains did not show significant difference to AFM1 binding ability as compared to live cells. Results of high performance liquid chromatography (HPLC) observed that W. confusa H1 and L. plantarum had shown 78% and 72% AFM1 binding in PBS at 37 °C after 72 h. There was a significant difference (P < 0.05) in AFM1 binding ability of L. plantarum S2 at 37 °C & 4 °C, but such difference was not observed for W. confusa H1. Moreover, these isolates also observed reduction of naturally present AFM1 in 10% RSM. This is the first study reporting the AFM1 binding capability of the genus Weissella (strain W. confusa H1) and Enterococcus durans. These results showed that indigenous strains of LAB are able to bind AFM1 effectively.

In vitro characterization of antimicrobial activity of an endophytic bacterium Enterobacter cloacae (MG001451) isolated from Ocimum sanctum

Enterobacter cloacae (MG001451) isolated from surface-sterilized parts of Ocimum sanctum showed potent antimicrobial activities against six bacterial human pathogens (Staphylococcus aureus, Escherichia coli, Bacillus subtilis, Klebsiella pneumonia, Salmonella enterica, Pseudomonas aeruginosa), one bacterial phytopathogen (Xanthomonas oryzae) and two fungal phytopathogens (Pythium debaryanum and Rhizoctonia solani). The antibacterial activities were studied through Disc diffusion, Bacteriocin Like Inhibition Studies BLIS and Agar cup plate method and confirmed through Thin Layer Chromatography (TLC)-Bioautography. The Minimum Inhibitory Concentration (MIC) value of the extract of E. cloacae against the bacterial test organisms ranged from 0.39–12.5 mg/ml. Lower MIC and Minimum Microbicidal Concentration (MMC) values were found against Bacillus subtilis followed by Staphylococcus aureus. The isolate E. cloacae also inhibited the growth of Rhizoctonia solani and Pythium debaryanum, while studied through, dual culture and well plate assay method. Higher inhibition % was reported against Rhizoctonia solani(73.75 ± 2.5%) than Pythium debaryanum (60.24 ± 2.08%). Secondary metabolites like alkaloids, phenols, polychlorinated biphenyls, aromatic hydrocarbons, esters, etc. were identified in the extract through Gas Chromatography coupled Mass Spectrometry (GC-MS), which could be responsible for antimicrobial activities as observed during the investigation.

Application of a binary mixture of bio-extract on organic cotton fabric

Rosmarinus officinalis and Psidium guajava plant leaves are having rich sources of natural nutrients and phytochemical compounds. These phytochemical compounds inhibit the growth of microorganisms present in fewer animals and the human body. In this study, wound healing property was developed on organic cotton fabric using an equal ratio of Rosmarinus officinalis and Psidium guajava extract by following the pad-dry-cure method. The presence of phytochemical compounds was ascertained through Thin Layer Chromatography (TLC) and the existence of the functional group in the herbal extract was analyzed by Fourier Transform Infrared Spectroscopy (FTIR). Microcapsules are prepared from the herbal extract using an ionic gelation process and applied to organic cotton fabric. The size of the particle present in the microencapsulated extract was confirmed using Scanning Electron Microscope. The antibacterial activity of the herbal extract and treated cotton fabric was evaluated using the agar well diffusion and parallel streak method. A high zone of inhibition attained against Escherichia coli and Staphylococcus aureus indicates antibacterial potency of the extract. The thickness, stiffness, air permeability, vertical wicking length, and water vapour permeability properties were marginally affected after treatment. The wound healing analysis of the treated fabrics was carried out using an in-vitro wound healing scratch assay.

Multivariate regression and artificial neural network to predict phenolic content in Schinus terebinthifolius stem bark through TLC images

This study aims to determine a suitable artificial neural network (ANN) regression model for quantification of polyphenols from herbal matrices through thin-layer chromatography (TLC) images. Gallic acid was used as a standard for the development and evaluation of this analytical approach. Analyses were performed on samples extracted from the stem bark of Schinus terebinthifolius, an important species in traditional Brazilian medicine. Calibration curves were obtained by TLC for image acquisition and assessment. High-performance liquid chromatography was used to verify the relationship between the data. The image features were associated with artificial intelligence techniques to determine an ANN configuration. This approach provides an innovative and low-cost analytical strategy with satisfactory performance for the prediction of phenolic content through TLC plate images.

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

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