TLC Analysis Research Articles

Discovery and characterization of an FAD-dependent glucose 6-dehydrogenase (74 characters including spaces).

Many patients with diabetes use self-measurement devices for blood glucose to understand their blood glucose levels. Most of these devices utilize FAD-dependent glucose dehydrogenase (FAD-GDH) to determine blood glucose levels. For this purpose, FAD-GDHs specifically oxidizing glucose among the sugars present in blood is required. Many FAD-GDHs with high substrate specificity have been reported previously; however, their substrate specificity is insufficient as they also react with xylose. Therefore, we aimed to identify FAD-GDHs without xylose reactivity. We screened and obtained a new enzyme from Colletotrichum plurivorum (CpGDH). CpGDH showed high activity to glucose in the presence of electron mediators but low activity to xylose. We prepared the glucose oxidation products using CpGDH and subjected to TLC, HPLC, MS, and NMR analyses. The results demonstrated that CpGDH is a previously unknown FAD-dependent glucose 6-dehydrogenase (FAD-G6DH) that oxidizes glucose to glucuronic acid. The stoichiometric ratio of the substrate and electron mediator was 1:2, suggesting that CpGDH catalyzes two-step oxidation reactions, including oxidation of primary alcohols to aldehydes and of aldehydes to carboxylic acids. We concluded that CpGDH has the unique substrate-binding manner based on the result of docking simulation of CpGDH with a substrate glucose. We then constructed a phylogenetic tree of carbohydrate-related flavoproteins including FAD-G6DHs, indicating that FAD-G6DHs are different from the known FAD-dependent oxidoreductases. Overall, this study is the first to report FAD-G6DHs. These results will likely contribute to the development of more accurate blood glucose sensors and further research on the metabolisms of glucosides and their metabolites.

Carob leaves: Phytochemistry, antioxidant properties, vasorelaxant effect and mechanism of action.

Ceratonia siliqua L., is a species of significant nutritional and industrial interest with extensive traditional uses. This fabaceae is renowned for its medicinal properties, including the treatment of high blood pressure. Due to its chemical composition, carob exhibits several valuable therapeutic functions such as antioxidant, antidiarrheal, antidiabetic, and antibacterial actions. This study investigates the chemical composition of Ceratonia siliqua L. leaves aqueous extract (CsAE) and explores the vasorelaxant effect and its underlying mechanisms. Acute toxicity and antioxidant activity of CsAE were also examined. The phytochemical profile was elucidated using TLC and UHPLC-MS. The vasorelaxant effect and mechanisms were studied on thoracic aortic rings from normotensive rats, using various antagonists. Acute toxicity was assessed by orally administering the extract to mice. Antioxidant activity was evaluated using β-carotene bleaching and DPPH. TLC analysis of CsAE reveals flavonoids and hydrolysable tannins. Gallic acid, myricitrin, quercitrin as well as galloylglucopyranoside derivatives were identified by UHPLC-MS. CsAE relaxed phenylephrine-precontracted aorta in a concentration-dependent manner. This response was reduced when the aorta was denuded or pretreated with L-NAME, hydroxocobalamin, ODQ, 4-AP, TEA, calmidazolium chloride, and thapsigargin. CsAE showed significant antioxidant activity with no observed toxicity in the experimental animals. CsAE has a significant vasodilatory effect, mediated through the CaM/eNOS/sGC pathway, activation of Kca and Kv, and intracellular calcium mobilization into SERCA. It also exhibits strong antioxidant activity, with no observed toxicity in the experimental animals. These findings represent the first evidence of the vasorelaxant effect of Ceratonia siliqua L. leaves from Eastern Morocco.

Phenolic Derivatives Isolated from Chamaecrista hildebrandtii (Vatke) Lock and Their Biological Activities

Chamaecrista hildebrandtii is used to prepare herbal medicines as laxative and purgative agents to treat diseases such as diarrhea, stomach ulcers, healing wounds and skin infections. Thus, this plant has shown great interest to researchers, but the chemical constituent of this plant has not been determined. The objective of the study was the extraction and purification of phytochemicals from C. hildebrandtii crude leaf extracts and to determine their antioxidant and antimicrobial ability. Extraction of the C. hildebrandtii leaves was done by macerating shade-dried and milled leaves in methanol that was further partitioned using water, hexane and ethyl acetate. Ethyl acetate fraction underwent further TLC analysis and repeated column chromatography with various solvent combinations to give a total of twelve fractions. Spectroscopic analysis was done using 1H NMR, 13C NMR, 1H-1H COSY, 2D-HSQC and 13C DEPT-135 and carefully compared with the literature available. Two phenolic compounds emodin (1,3,8-trihydroxy-6-methylanthraquinone) and chrysophanol (1,2-dihydroxy-6-methylanthrquinone) were isolated and identified. The antimicrobial activity was tested by disc diffusion method using Mueller Hinton agar growth medium. Emodin exhibited comparatively higher inhibitory activity against microbes in comparison with chrysophanol. Emodin showed a 55.1% DPPH inhibition while chrysophanol recorded a 58.2% inhibition. Emodin showed a desirable IC50 of 7.3 mg/mL compared to chrysophanol with an IC50 value of 12.3 mg/mL. The IC50 values of the two isolates were slightly higher than that of ascorbic acid (IC50 = 3.24 mg/mL). The outcome revealed that C. hildebrandtii leaf extracts may represent a promising natural source of polyphenols, which can be used as antimicrobial and antioxidant agents.

Lichenase and Cellobiohydrolase Activities of a Novel Bi-Functional β-Glucanase from the Marine Bacterium Streptomyces sp. J103.

In this study, we report the molecular and enzymatic characterisation of Spg103, a novel bifunctional β-glucanase from the marine bacterium Streptomyces sp. J103. Recombinant Spg103 (rSpg103) functioned optimally at 60 °C and pH 6. Notably, Spg103 exhibited distinct stability properties, with increased activity in the presence of Na+ and EDTA. Spg103 displays both lichenase and cellobiohydrolase activity. Despite possessing a GH5 cellulase domain, FN3 and CBM3 domains characteristic of cellulases and CBHs, biochemical assays showed that rSpg103 exhibited higher activity towards mixed β-1,3-1,4-glucan such as barley β-glucan and lichenan than towards beta-1,4-linkages. The endolytic activity of the enzyme was confirmed by TLC and UPLC-MS analyses, which identified cellotriose as the main hydrolysis product. In addition, Spg103 exhibited an exo-type activity, selectively releasing cellobiose units from cellooligosaccharides, which is characteristic of cellobiohydrolases. These results demonstrate the potential of Spg103 for a variety of biotechnological applications, particularly those requiring tailor-made enzymatic degradation of mixed-linked β-glucans. This study provides a basis for further structural and functional investigations of the bifunctional enzyme and highlights Spg103 as a promising candidate for industrial applications.

Halogenated Cobalt Bis-Dicarbollide Strong Acids as Reusable Homogeneous Catalysts for Fatty Acid Esterification with Methanol or Ethanol

The most commonly used homogeneous catalyst for fatty acid esterification is a corrosive sulphuric acid. However, this requires costly investment in non-corrosive equipment, presents a safety risk, is time consuming, and increases effluent generation. In this study, inorganic 3D heteroborane cluster strong acids are employed for the first time as homogeneous catalysts. Three novel isomeric tetrachlorido and tetrabromido derivatives of 3,3′-commo-bis[undecahydrido-closo-1,2-dicarba-3-cobaltadodecaborate](1−) [1−] were synthesised and fully characterised using a range of analytical techniques, including NMR, TLC, HPLC, MS, UV-Vis, melting point (MP), CHN analyses, and XRD. Ultimately, H3O[8,8′-Cl2-1−] was identified as the most efficient, reusable, and non-corrosive homogeneous catalyst for the esterification of four fatty acids. The reactions are conducted in an excess of alcohol at reflux. The effective absorption of water vapour provided by the molecular sieves maximises acid conversion. The hydrophobic dye Sudan black B was employed as an acid-base indicator to facilitate a comparison of the H0 acidity function of sulphuric acid and halogenated heteroboranoic acids when dissolved together in methanol. The 23Na NMR analysis demonstrated that the application of dry methanol resulted in the displacement of Na+ ions from zeolite, which subsequently exchanged the H3O+ ions of the acid. This process led to a gradual reduction in the efficiency of the catalysts, particularly with repeated use. The solution to this issue is to regenerate the catalyst on the ion exchanger following each reaction. In contrast to the published methods, our new approach meets 10 of 12 green chemistry principles.

Screening for substandard and falsified medicines in Nigeria using visual inspection and GPHF-Minilab analysis: lessons learnt for future training of health workers and pharmacy personnel.

Substandard and falsified (SF) medicines are a serious threat to public health in low- and middle-income countries (LMICs). Visual inspection of medicines and screening analysis using the Global Pharma Health Fund (GPHF)-Minilab are important in medicine quality surveillance in low-resource settings. Recently, 260 medicine samples from Nigeria had been investigated for assay and dissolution according to the United States Pharmacopeia (USP). In the present study, these results were compared to the results of the investigation of the same samples by visual inspection and by GPHF-Minilab analysis by local personnel in Nigeria. Visual inspection identified many deficiencies of dosage units and packaging information in SF medicines. All four falsified medicines were readily identifiable, primarily from serious spelling errors in the labelling, and from manufacturer names which could not be verified using internet resources. In GPHF-Minilab disintegration testing, two samples did not disintegrate even after 60 min; both were found to fail USP dissolution testing with extreme deviations. Of the 20 samples which deviated in USP assay analysis by more than 20% from the declared API amount, seven (35%) were detected as non-compliant in TLC analysis. Evaluation by TLC image analysis with a recently developed smartphone application (named TLCyzer) increased sensitivity to 62.5% but led to an unacceptably low specificity (75.2%). Additional training of the local personnel improved the results of both TLC analysis and TLCyzer evaluation. Photographs of the visual deficiencies and of the TLC analysis results of the SF medicines are provided as PowerPoint and PDF slides with this publication, for future training courses of pharmacy staff and health workers in LMICs. Visual inspection, and screening analysis with simple, rapid and inexpensive methods, are important in the surveillance for SF medicines in LMICs. This study provides data on the potential and the limitations of such screenings.

Discovery of EDA‐Complex Photochemical Reactions Using Computational Vision

Herein, we report a multidimensional screening strategy to discover Electron Donor‐Acceptor (EDA) complex photochemical reactions using only simple photographic devices, such as webcams or cellphones, combined with TLC analysis. An algorithm was designed to automatically identify EDA‐complex reactive mixtures in solution by processing digital images from a 96‐well microplate, alongside TLC results. The code detects the absorption region of the mixture in the visible spectrum and quantifies color changes through grayscale values. Integrated automatic TLC analysis further classifies mixtures as colorimetric reactions, non‐reactive, or potentially reactive EDA complexes. Leveraging this screening platform, we have developed a new EDA‐mediated approach to synthesize iminophosphoranes in yields up to 90%.

Quality Evaluation of Kidney Tea Granules From Different Origins Based on TLC, HPLC Fingerprinting, and Quantitative Analysis Combined With Chemical Pattern Recognition.

Kidney tea is an essential herbal medicine. It is widely used to treat conditions such as urinary stones, gallstones, and rheumatoid arthritis. There is currently no standardized or widely accepted research strategy for evaluating the quality of kidney tea granules (KTGs) after granulation. In this study, we aim to establish a comprehensive strategy for evaluating the quality of KTGs produced from different sources of kidney tea. A TLC combined with HPLC method was established to identify the chemical components in KTGs, and HPLC method was used to determine the contents of rosmarinic acid of KTGs. In order to distinguish samples and identify differential components, similarity analysis, hierarchical cluster analysis (HCA), principal component analysis (PCA), and orthogonal partial least squares discriminant analysis (OPLS-DA) were conducted. TLC and HPLC detection confirmed three chemical components of KTGs, which are rosmarinic acid, caffeic acid, and lithospermic acid. HPLC fingerprint analysis revealed a total of seven common peaks in 15 batches of KTGs. Similarity analysis showed that the similarity of all 15 batches of KTGs was greater than 0.969. The peak areas of the seven common peaks were identified by chemical pattern recognition, and the results showed that most of the KTGs from different origins were clustered together, with small differences between them. The PCA and OPLS-DA results showed that two principal components can represent 82.638% of the common peaks of KTGs, among which peak 5 represents rosmarinic acid, which is the main differential biomarker of KTGs from different regions. Quantitative analysis of rosmarinic acid in KTG samples was performed using HPLC fingerprint conditions and the content of rosmarinic acid in 15 batches of KTGs samples was measured to be between 8.01-14.61 mg/g. This study combines TLC, HPLC, and chemometrics to establish a stable and reliable method that can quickly and effectively identify the components of KTGs, accurately quantify known components in KTGs, and provide reference for the quality evaluation of KTGs.

Expression and characterization of a novel halophilic GH10 β-1,4-xylanase from Trichoderma asperellum ND-1 and its synergism with a commercial α-L-arabinofuranosidase on arabinoxylan degradation

Enzymatic hydrolysis of arabinoxylan is of cost-effective strategy to yield valuable macromolecules, e.g., xylooligosaccharides (XOS). A novel halophilic GH10 xylanase (TaXYL10) from Trichoderma asperellum ND-1 was over-expressed in Pichia pastoris and migrated as a single band (~36 kDa) in SDS-PAGE. TaXYL10 displayed >80 % activity in the presence of 4.28 M NaCl and 10 % ethanol. Moreover, TaXYL10 exhibited optimal activity at pH 6.0 and 55 °C, and remarkable pH stability (>80 % activity at pH 4.0–6.0). K+ and Al3+ could remarkably promote TaXYL10 activity, while the presence of 10 mM Fe2+, Zn2+, Cu2+ and Fe3+ decreased its activity. TaXYL10 possesses the highest catalytic activity towards beechwood xylan. TLC analysis revealed that it could rapidly degrade xylan and XOS with DP ≥ 3, yielding xylotriose and xylobiose. Site-directed mutagenesis indicated that Glu154 and Glu259 are crucial active residues for TaXYL10, while Asp295 and Glu69 played auxiliary roles in xylan hydrolysis. Additionally, TaXYL10 acted cooperatively with a commercial α-L-arabinofuranosidase (AnAra) towards arabinoxylan degradation (583.5 μg/mL), a greater synergy degree of 1.79 was obtained after optimizing enzymatic ratios. This work not only expands the diversity of Trichoderma GH10 xylanases, but also reveals the promising potential of TaXYL10 in various industrial applications.

Phytochemical Constituent and Cumulative or Antagonistic Effects of Crops Plant Organ Combination on Free Radical Scavenging Capacity and Antioxidant Compound Content

Aims: In Burkina Faso, the majority of plant-based recipes are a combination of plant organs from the same or different species. For these recipes’ producers, the aim is to improve efficiency. However, our ethnobotanical surveys have revealed that for some species, efficacy is better when the species is not combined, or when it is combined in small proportions with other species. The aim of this study is to assess the effect of plant organ combination on free radical scavenging activity, phenolic compound content and flavonoid content. Study Design: The work combined harvesting and processing of plant material from September to November 2023, and laboratories analysis from January to March 2024 at Natural Substances Departement of Institut de Recherche en Sciences Appliquées et Technologie, Ouagadougou. Methodology: We prepared two batches of recipes, first combining two species from the same genus, then two species from different genera. The powder of each plant without combination was also used to compare data. Phytochemical screening was first carried out with TLC and LC-MS analysis on each plant extract. We then assessed the phenolic compound content (PCC) using Folin-Ciocalteu method, flavonoid content (FCC) using aluminum chloride test, antioxidant content using TEAC method and free radical scavenging activity using DPPH method, for different batches. Results: Phytochemical constituent in the three species are phenolic compounds, terpenoids and nitrogen derivatives. The results of biological property evaluation suggest that for best efficacy and to obtain a high phenolic compound content, Lippia multiflora leaves should be used separately, or combined in 70% proportions with Lippia alba, then in 50% proportions if the combination is made with Ocimum basilicum. Conclusion: This research has provided relevant results to guide players in the field of traditional medicine and users of herbal remedies.

Improvement of 9α-hydroxyandrost-4-ene-3,17-dione production in Mycolicibacterium neoaurum by regulation of cell wall formation and transcriptional regulator PadR

The biotransformation of phytosterol into high value steroid intermediates such as 9α-hydroxyandrost-4-ene-3,17-dione (9-OHAD) in Mycolicibacterium is the cornerstone of the steroid pharmaceuticals. However, the limited permeability of the dense mycobacterial cell wall severely hinders the efficient transportation of phytosterol and their bioconversion to 9-OHAD. In this study, we disrupted the genetic pathways involved in trehalose biosynthesis, trehalose recycle and by-product formation, leading to alterations in cell wall formation, cell permeability and 9-OHAD productivity. This manipulation led to an increase of 63.7% in the yield of 9-OHAD, reaching 10.8 g/L at a phytosterol concentration of 20 g/L in shake flask. The enhancement of cell permeability and 9-OHAD production were achieved through the deletion of genes TreS, TreY, OtsA, LpqY, and SugC, as well as the inactivation of regulator PadR. Notably, it was found that the increase in TMM content of cell wall components via TLC analysis directly affected the distribution of 9-OHAD within and outside the cell, ultimately leading to an increase in extracellular production of 9-OHAD from 12% to 32.1%. Therefore, this study provides with an effective strategy for enhancing 9-OHAD production by increasing cell permeability while minimizing by-product 4-AD formation.

One-pot biocatalysis of potato rhamnogalacturonan and the role of its deacetylation in efficient inhibition of colon cancer cells and hydrogel mediated colon-targeted drug delivery

Deacetylation of potato rhamnogalacturonan (PRG) by rhamnogalacturonan acetyl esterase (CtPae12B) was explored for enhanced hydrolysis of PRG by rhamnogalacturonan lyase (CtRGLf) and the effects of deacetylated PRG were studied in enhancing inhibition of colon-cancer cells and formation of colon-targeting drug delivery material. Pre-treatment of PRG with CtPae12B resulted in increased relative activity of CtRGLf. CtPae12B removed acetyl groups from both O-2 and O-3 positions of D-galactopyranosyluronic acid residues of PRG, resulting in 98 % deacetylation. PRG displayed 21.9 % degree of acetylation and 7.7 % degree of methylation. TLC and ESI-MS analysis of CtRGLf hydrolysed PRG showed unsaturated RG di-saccharide as the smallest product, with m/z 322. Deacetylated PRG-oligosaccharides displayed higher, 50 % inhibition of colon-cancer HCT-116 cells (with shrunken and globular morphology) than 35 % inhibition by acetylated PRG-oligosaccharides. FESEM and BET analysis of CtPae12B-treated PRG showed porous structure and significantly higher total surface area and pore volume than non-enzyme treated PRG. Higher drug entrapment efficiency and lower drug release rate of CtPae12B-treated PRG hydrogel (0.0033 min−1 at pH 1.2 and 0.009 min−1 at pH 7.4), than non-enzyme treated PRG hydrogel, (0.0057 min−1 at pH 1.2 and 0.02 min−1 at pH 7.4), showed it to be a potential biomaterial for sustainable colon-targeted drug delivery.

Spasmolytic Effects Of Kauranoid And Ester Of Linear Alkaloid From The Aerial Part Of Croton Borarium (Euphorbiaceae) Endemic In South Part Of Madagascar

The aerial part of the plant known under the vernacular name Somorombohitse (Malagasy name) and scientifically named Croton Borarium (Euphorbiaceae) endemic in the South part of Madagascar, is used by the local communities to treat asthma and hypertension. The biologic test result riddling of the isolated trachea of guinea pig stimulated by the acetylcholine in 210-2M showed antiasthmatic activity with the CE50= 0.85±0.001µg/ml (n=6). Effet concentration of carbachol from 10-9M to 10-3M have been realized in absence and in presence of the hydro-ethanolic extract of Cr. Borarium to 0.1mg/ml, verapamil to 10µM and atropine to 10nM. The relaxing activity of hydro-ethanolic extract from 0.1 to 3 mg/ml has been executed on the isolated aorta and jejunum of rabbit stimulated with the KCl to 50nM. White the using of the extract technic by partitioned liquid-liquid to permitted localizing the active fraction (Hex/DCM: 3/1) with CI50= 0.275±0.003µg/ml (n=6).The bioassay-guided fractionation of the active fraction using repeated silica gel column chromatography resulted in the isolation of two pure compounds active as evidenced by analytical TLC and HPLC analysis. The elucidation of pharmacologic mechanism of these active principles has showed that, to first and active principle bring into play the calcic movement through the cellular membrane of vascular and polished muscle and the second is the by way of NO intermediate. Their chemical structures of two pure compounds were determined by 1D and 2D NMR spectroscopy and spectroscopy High-resolution MS.

Evaluation of antimalarial compounds in Prosopis africana stem bark fractions against Plasmodium berghei

Prosopis africana, a traditional medicinal plant for malaria treatment, requires further scientific evidence. This study aimed to assess the antimalarial potential of Prosopis africana stem bark fractions in Plasmodium berghei-infected mice and identify the responsible compounds. The aqueous extract underwent partitioning with n-hexane, ethyl acetate, and methanol, followed by antimalarial evaluation. F5 was the most potent fraction based on its antimalarial potential. The study pooled 36 chromatographic separation sub-fractions into five using TLC analysis Rf values. In Plasmodium berghei-infected mice, fraction F5 at 6.25, 12.5, 25, and 50 mg/kg body weight (bw) was tested for antimalarial activity. Compound analysis of F5 utilized ultra-performance liquid chromatography-quadrupolar mass spectrometry. Statistical analysis employed Analysis of Variance and Duncan's multiple range tests (p < 0.05). The methanol, n-hexane, and ethylacetate extracts displayed suppression of P. berghei by 90%, 72.2%, and 73.6%, respectively. Methanol, n-hexane, and ethylacetate extracts suppressed P. berghei by 90%, 72.2%, and 73.6% respectively. F5 showed significantly higher chemo-suppression (67.21%) than F1 (63.94%), F2 (6.99%), F3 (2.81%), and F4 (37.3%). At 12.5 mg/kg bw, F5 suppressed P. berghei by 91.09%, surpassing chloroquine (87.79%), and restored altered hematological indices and CD4+ counts. Compounds identified in F5 included caffeate, linalool, terpinene-4-ol, quinovic acid, quercetin, prosopinine, catechin, and apigeninin. In conclusion, the fraction F5 of P. africana stem bark exhibited higher antimalarial activity than chloroquine and the identified compound might likely contribute to its activity.

Ginsenoside Profile of White Ginseng Roots and Tinctures

Ginseng roots are medicinal plant raw materials used in pharmaceutical practice in a number of countries due to their pharmacological activities, including tonic. Ginsenosides of ginseng are the main group of biologically active substances responsible for the pharmacological effects of ginseng.One of the modern methods for analyzing ginsenosides is UPLC-MS/MS, which allows one to gain a broad understanding of the ginsenoside profile. The authors used TLC analysis and UPLC-MS/MS to identify and quantify ginsenosides. Of the 18 identified ginsenosides, ginsenosidesRb 1, Rc, Re, Rg1 can be considered as marker ones. These approaches can serve as the basis for the development of regulatory documentation for ginseng and medicines based on it. The total content of ginsenoside in tinctures is higher than in the medicinal plant material of white ginseng roots. Chemical compounds studied in this article: Ginsenoside Rb1 (PubChem CID: 9898279); Ginsenoside Rb2 (PubChem CID: 6917976); Ginsenoside Rb3 (PubChem CID: 12912363); GinsenosideRc (PubChem CID: 12855889); Ginsenoside Rd (PubChem CID: 24721561); Ginsenoside Re (PubChem CID: 119307); Ginsenoside Rf (PubChem CID: 441922); Ginsenoside Rg1 (PubChem CID: 441923); Ginsenoside S-Rg2 (PubChem CID: 12912322); Ginsenoside Ro (PubChem CID: 11815492); Ginsenoside S-Rg3 (PubChem CID: 9918693); Ginsenoside Rg5 (PubChem CID: 11550001); Ginsenoside R-Rh1 (PubChem CID: 21599923); Ginsenoside S-Rh2 (PubChem CID: 91668381); Ginsenoside Rk1 (PubChem CID: 11499198); Ginsenoside Rk3 (PubChem CID: 75412555); Ginsenoside F2 (PubChem CID: 9918692); Ginsenoside F3 (PubChem CID: 46887678); Ginsenoside F4 (PubChem CID: 73717457); Ginsenoside CK (PubChem CID: 9852086).

3,4-Dihydropyrimidine-2(1H)-one/thione Derivatives as Anti-inflammatory and Antioxidant Agents: Synthesis, Biological Activity, and Docking Studies

aims: A series of sixteen new 4-(substituted phenyl) - 6-(substituted phenylamino)-3, 4-dihydropyrimidine-2(1H)-ones/thiones derivatives have been synthesized in two step reaction. background: Free radicals are widely known to play a significant part in the inflammatory process. Many non-steroidal anti-inflammatory medicines have been shown to work as radical scavengers or as inhibitors of free radical generation. Inflammation is a part of the vascular tissues' complicated biological reaction to adverse stimuli like bacteria, damaged cells, or irritants. The organism's protective response to damaging stimuli is inflammation, which aims to get rid of them and start the healing process. The inflammatory response has several advantageous effects, such as stopping the spread of dangerous substances to nearby tissues, getting rid of cell waste and infections, and getting the body ready for repair objective: To synthesize new derivatives as anti-inflammatory and antioxidant agents. method: In first step chalcones (I1-I6) was obtained and in the second step, these chalcones were reacted with urea and thiourea in the presence of potassium hydroxide to obtain the corresponding pyrimidinones and thiopyrimidinones. The structures of the synthesized compounds 4-(substituted phenyl)-6-(substituted phenylamino)-3,4-dihydropyrimidine-2(1H)-ones/thione derivatives are investigated by means of TLC (visualized it in Iodine chamber), IR, mass, 1H-NMR spectral and elemental analysis. The title compounds evaluated for anti-inflammatory as well as antioxidant activity. Anti-inflammatory activity in vivo was carried out by carrageenan induced rat-paw edema method and compared with standard drug diclofenac sodium and antioxidant activity was measured by DPPH, FRAP and Hydrogen peroxide (H2O2) method and compared with standard ascorbic acid. result: Electron donating groups showed better antioxidant activity as compared to electron withdrawing ones in both activities. All the compounds exhibited good to moderate activity. Compound DHPM8 shows better antioxidant activity and compound DHPM6 shows better anti-inflammatory activity while compound DHPM11 shows minimum antioxidant as well as anti-inflammatory activity than other compounds. conclusion: To better understanding, we have performed molecular docking simulation. The molecular docking studies revealed that all synthesized compound were showed good receptor binding interaction in which compound 9 had highest docking score of 9.8 due to interaction with CYS36, PRO153, TYR130, GLY45, LEU152, ARG469, LYS468 and GLU465. other: NA

Synthesis, Computational Studies, and Biological Evaluation of Sulpha-methoxazole-Based Schiff Bases as Antimicrobial Agents

Background: Sulphamethoxazole-based Schiff-base compounds display poten-tial antibacterial and antifungal activity. Sulphamethoxazole is considered to be a versatile pharmacophore that can be utilized for designing and developing numerous bioactive lead compounds. In this work, some new sulphamethoxazole-based Schiff base compounds were synthesized, which are expected to possess antimicrobial activity, making them potentially useful for treating microbial infections. Objective: Concerning issues of drug resistance in presently available drugs, this study aimed to synthesize new sulphamethoxazole-based Schiff bases and evaluate their antimi-crobial activity. Methods: New sulphamethoxazole-based Schiff bases were synthesized by condensing sul-phamethoxazole with various acetophenones in methanol in the presence of glacial acetic acid. The synthesized compounds were characterized using various techniques, such as TLC, melting point, IR, NMR, and mass analysis. The computational properties of the com-pounds were also assessed using online software programs, and the similarity of the target compounds was also calculated as compared to sulphamethoxazole and clotrimazole. The antimicrobial activity of the target compounds was tested against Bacillus subtilis (Gram-positive), Escherichia coli (Gram-negative), and Candida albicans. Results: The target compounds (3a-f) were successfully synthesized and characterized by spectroscopic and analytical methods. The results of computational properties, similarity, and antimicrobial activity against B. subtilis, E. coli, and C. albicans of new sulphamethox-azole-based Schiff bases showed significant antimicrobial potential. result: The results of computational properties, similarity and antimicrobial activity against B. subtilis, E. coli and C. albicans of new sulphamethoxazole based Schiff bases showed significant antimicrobial potential. Conclusion: The synthesized new Schiff bases, particularly compound 3c, exhibited prom-ising antimicrobial activity and good physicochemical properties as compared to standard drugs, indicating their potential for further development as antimicrobial agents.

Phenazines are involved in the antagonism of a novel subspecies of Pseudomonas chlororaphis strain S1Bt23 against Pythium ultimum

Long-term use of chemical fungicides to control plant diseases caused by fungi and oomycetes has led to pathogen resistance and negative impacts on public health and environment. There is a global search for eco-friendly methods and antagonistic bacteria are emerging as alternatives. We isolated a potent antagonistic bacterial strain (S1Bt23) from woodland soil in Québec, Canada. Taxonomic characterization by 16S rRNA, multi-locus sequence analysis, pairwise whole-genome comparisons, phylogenomics and phenotypic data identified strain S1Bt23 as a novel subspecies within Pseudomonas chlororaphis. In dual culture studies, strain S1Bt23 exhibited potent mycelial growth inhibition (60.2–66.7%) against Pythium ultimum. Furthermore, strain S1Bt23 was able to significantly bioprotect potato tuber slices from the development of necrosis inducible by P. ultimum. Annotations of the whole genome sequence of S1Bt23 revealed the presence of an arsenal of secondary metabolites including the complete phenazine biosynthetic cluster (phzABCDEFG). Thin-layer (TLC) and high-performance liquid (HPLC) chromatographic analyses of S1Bt23 extracts confirmed the production of phenazines, potent antifungal compounds. CRISPR/Cas9-mediated deletion of phzB (S1Bt23ΔphzB) or phzF (S1Bt23ΔphzF) gene abrogated phenazine production based on TLC and HPLC analyses. Also, S1Bt23ΔphzB and S1Bt23ΔphzF mutants lost antagonistic activity and bioprotection ability of potato tubers against P. ultimum. This demonstrated that phenazines are involved in the antagonistic activity of S1Bt23 against P. ultimum. Finally, based on genotypic and phenotypic data, we taxonomically conclude that S1Bt23 represents a novel subspecies for which the name Pseudomonas chlororaphis subsp. phenazini is proposed.

HPTLC sequentially coupled to UV and SERS: A cost-effective tool for confirmative identification and quantitation of sildenafil in falsified herbal products

The detection of falsified drugs usually requires multi-disciplinary analysis for confirmative identification. Among hyphenated techniques with high specificity detection, thin-layer chromatography coupled with surface-enhanced Raman spectroscopy (TLC-SERS) is an efficient choice, especially for herbal products with diversified matrix. In this study, HPTLC was coupled to two detection techniques: UV absorption and Raman scattering with silver colloid enhancement for the analysis of sildenafil adulterated in herbal products. With this approach, orthogonal UV and SERS spectral data was collected, so that confirmative results could be obtained within a single TLC analysis. How this approach helped to reduce chances of false positive or false negative results was also discussed. The HPTLC sequentially coupled to UV and SERS (HPTLC-UV-SERS) method was developed and validated parallelly on the UV and SERS signals. To improve the repeatability of the SERS signal, several analytical conditions were optimized, so that direct quantitation with TLC-SERS was feasible without chemometric data extrapolation. The determination was done with UV scanning at 304 nm for HPTLC and with SERS signal at 1580 cm−1 (excitation 633 nm). The TLC-SERS method had a detection limit of 1.65 ng/spot, 95 times lower than HPTLC method (157 ng/spot). The HPTLC-UV-SERS method was applied on 24 real herbal samples collected from the market, among which 3 real samples were positive to sildenafil, and quantitation results by UV and SERS were in consistency. Not only this method was proved feasible for practical applications, but the recommendations for TLC-SERS procedures could also be useful in TLC-SERS method development for other compounds.

Identification of the Secondary Metabolites of Sargassum Tenerrimum and their Molecular Docking Analysis against the Targets of Anxiety, Depression and Cognitive Disorder

Objective: This article aimed to identify the bioactive compounds present in the brown algae Sargassum tenerrimum using TLC and HPTLC fingerprinting analysis and followed in silico molecular docking against a potential target of anxiety, depression, and cognitive disorder with identified compounds. Methods: Bioactive compounds were identified from the methanolic extract of Sargassum tenerrimum through TLC and HPTLC fingerprinting analysis. In silico molecular docking against a potential target of anxiety, depression, and cognitive disorder was performed on the latest version of AutoDock Vina v.1.2.0 software. The pharmacokinetic profile and possible bioactivities of the compounds were predicted using SwissADME. Results: Fucoxanthin, β-Cryptoxanthin, and Canthaxanthin were identified from the brown algae Sargassum tenerrimum through TLC and HPTLC fingerprinting analysis. Fucoxanthin showed the highest fitness score of -9.7 kcal/mol, -9.6 kcal/mol, and -9.7 kcal/mol against the target protein GABA-A, 5ht2c, and AchE, respectively. β-Cryptoxanthin showed the highest fitness score of -9.4 kcal/mol against target SERT compared with Fucoxanthin and Canthaxanthin. Canthaxanthin exhibited the highest fitness score- 7.5 kcal/mol, -9.0 kcal/mol, -9.7 kcal/mol, -9.1 kcal/mol, -9.1 kcal/mol, -7.4 kcal/mol, -7.9 kcal/mol and - 7.6 kcal/mol against the target receptor trkB, 5ht1A, D2, DAT, MOA-A, COMT, NMDA and 7nAchR respectively on the comparing with Fucoxanthin and β-Cryptoxanthin. Conclusion: In silico docking and ADME analysis concluded that the canthaxanthin acted through various targets and was safer than the fucoxanthin and β-Cryptoxanthin. Hence, canthaxanthin can be the best potential compound in the therapy of neuropsychological disorders.