Thin-layer Chromatographic Technique Research Articles

Eco-friendly High Performance Thin Layer Chromatography Technique for Quantification of Kojic Acid in Bulk and Formulations

Eco-friendly High Performance Thin Layer Chromatography Technique for Quantification of Kojic Acid in Bulk and Formulations

Stability Indicating HPTLC Method for Estimation of Cubebin in Crude Drug and Marketed Formulations

Background: Cubebin is a naturally occurring lactone lignan isolated from the plant species Piper cubeba. It exhibits a range of biological activities, including anti-inflammatory, antimicrobial, and anticancer properties. This compound has garnered significant interest for its potential therapeutic applications in traditional and modern medicine. Aim: This study aimed to develop a novel stability-indicating High-Performance Thin Layer Chromatography (HPTLC) technique that is simple yet precise in estimating cubebin in crude drug and marketed formulations. Method: Cubebin was estimated using precoated TLC silica gel 60 F254 plate as a stationary phase and n-hexane: ethyl acetate: formic acid: methanol (6:3.5:0.5:0.3 v/v/v/v) as a mobile phase with the chamber saturation time of 20 mins. Key Findings: Cubebin was detected and quantified at the wavelength of 283nm. The Rf value of cubebin in standard solution, Piper cubeba extract, Dhanwantari Vati, and Khadiradi Vati was found to be 0.494±0.0015, 0.492±0.002, 0.490±0.0020, 0.499±0.001 respectively. The calibration curve exhibited linearity within the range of 3000-13000 ng per band with a correlation coefficient (r2) of Cubebin was 0.9968. The method developed was utilized for the quantitation of cubebin in Piper cubeba and both the marketed formulations. Conclusion: A novel HPTLC method was developed and validated in accordance to ICH guidelines for the analysis of cubebin in crude drug and marketed formulations. The method was found to be simple, precise, accurate, and specific. This method is suitable for quality control and stability testing of cubebin-containing formulations.

Simultaneous Estimation of Monoterpenes (Eugenol, Eucalyptol and R-Limonene) by HPTLC Densitometric Method and Its Validation.

Plant-derived monoterpenes have various pharmacological applications, but the problem lies with their detection and estimation. Hence, it is crucial to develop a robust technique to overcome this problem. This work was aimed at developing a swift, accurate and reproducible technique for the simultaneous detection of three monoterpenes, namely, eugenol, eucalyptol (1,8- cineole), and R-limonene, using the high-performance thin-layer chromatography technique, as per International Council of Harmonization (ICH) guidelines. The best results were obtained in the eluent mixture prepared by using hexane: toluene: ethyl acetate (6:3:1, v/v/v), and silica gel-coated aluminum thin layer chromatography (TLC) plates 60F254 as a stationary phase that produced very sharp and well-resolved symmetrical peaks for eugenol, eucalyptol, and R-limonene at Rf values of 0.47 ± 0.03, 0.56 ± 0.03, and 0.71 ± 0.03, respectively. The linear range for eugenol, eucalyptol, and R-limonene was 1 to 15 ng/ spot (r2 = 0.9906), 100 to 600 ng/spot (r2 = 0.99625), 50 to 300 ng/spot (r2 = 0.9873), respectively. Furthermore, the limit of detection (LoD) and limit of quantitation (LoQ) values for eugenol were obtained at 0.0129 and 0.0391 ng/spot, followed by eucalyptol (0.82 and 2.48 ng/spot) and R-limonene (0.594 and 1.8 ng/spot), respectively. To our knowledge, we are the first to report a quick and reproducible high-performance thin-layer chromatography (HPTLC) technique for the simultaneous determination of eugenol, eucalyptol (1,8-cineole), and R-limonene. This method can further be used for the quantification of these secondary metabolites in plant extracts, enriched fractions, and pharmaceutical formulations.

Development and validation of High-Performance Thin Layer Chromatographic methods for simultaneous determination of antibacterial pharmaceutical formulations containing Clindamycin phosphate.

The main objective of this research is to establish and authenticate the High-Performance Thin-Layer Chromatographic techniques for the simultaneous assessment of Clindamycin phosphate in combination with Tretinoin (formulation-1) and Clindamycin phosphate with Adapalene (formulation-2), both of which are antimicrobial combinations. In the developed HPTLC methods, for combination-1, the optimized mobile phase was a mixture of n-Hexane: Ethyl acetate: ACN: Methanol (5:2:2:1 by volume) with observed Rf values of 0.413 and 0.787 for Clindamycin phosphate and Tretinoin, respectively. Similarly, for combination-2, the mobile phase comprised n-Hexane: Diethyl ether: Dichloromethane: Acetic acid (4:4:2:0.1 by volume) with recorded Rf values of 0.407 and 0.559 for Clindamycin phosphate and Adapalene, respectively. The developed methods are successfully validated as per the regulatory guidelines. This approach proves to be significantly time-saving and costeffective, rendering it suitable for routine analyses of the formulations containing selected antibacterial active pharmaceutical ingredients and commercial formulations.

Simultaneous Estimation of Monoterpenes (Eugenol, Eucalyptol And R-Limonene) by HPTLC Densitometric Method and Its Validation .

Plant-derived monoterpenes have various pharmacological applications, but the problem lies with their detection and estimation. Hence, it is crucial to develop a robust technique to overcome this problem. This work was aimed at developing a swift, accurate and reproducible technique for the simultaneous detection of three monoterpenes, namely, eugenol, eucalyptol (1,8-cineole), and R-limonene, using the high-performance thin-layer chromatography technique, as per ICH guidelines. The best results were obtained in the eluent mixture prepared by using hexane: toluene: ethyl acetate (6:3:1, v/v/v), and silica gelcoated aluminum TLC plates 60F254 as a stationary phase that produced very sharp and well-resolved symmetrical peaks for eugenol, eucalyptol, and R-limonene at Rf values of 0.47 ± 0.03, 0.56 ± 0.03, and 0.71 ± 0.03, respectively. The linear range for eugenol, eucalyptol, and R-limonene was 1 to 15 ng/spot (r2 = 0.9906), 100 to 600 ng/spot (r2 = 0.99625), 50 to 300 ng/ spot (r2 = 0.9873), respectively. Furthermore, the LoD and LoQ values for eugenol were obtained at 0.0129 and 0.0391 ng/spot, followed by eucalyptol (0.82 and 2.48 ng/spot) and R-limonene (0.594 and 1.8 ng/spot), respectively. To the best knowledge, we are the first to report a quick and reproducible HPTLC technique for the simultaneous determination of eugenol, eucalyptol (1,8-cineole), and R-limonene. This method can further be used for the quantification of these secondary metabolites in plant extracts, enriched fractions, and pharmaceutical formulations.

Larvicidal potential of Trachyspermum ammi essential oil and Delphinium speciosum extract against malaria, dengue, and filariasis mosquito vectors

Mosquito-borne diseases, such as malaria, dengue, and Zika, pose major public health challenges globally, affecting millions of people. The growing resistance of mosquito populations to synthetic insecticides underscores the critical need for effective and environmentally friendly larvicides. Although chemical pesticides can initially be effective, they often lead to negative environmental consequences and health hazards for non-target species, including humans. This study aimed to evaluate the larvicidal effects of Trachyspermum ammi essential oil and Delphinium speciosum extract on the larvae of three major mosquito species: Aedes aegypti, Anopheles stephensi, and Culex quinquefasciatus. Mosquito larvae of Ae. aegypti, An. stephensi, and Cx. quinquefasciatus were reared under controlled laboratory conditions. The larvicidal activity of T. ammi essential oil and D. speciosum extract was evaluated through standard bioassays, using various concentrations of essential oils (10, 20, 40, 80, and 160 ppm) and extracts (160, 320, 640, 1280, and 2560 ppm) to determine the lethal concentration (LC50) values after 24 h of exposure. Fresh plant materials were collected, with the essential oil extracted via hydro-distillation, and the extract prepared using methanol solvent extraction. The chemical composition of T. ammi essential oil was examined using gas chromatography-mass spectrometry (GC-MS). Additionally, the preliminary analysis of the chemical compounds in D. speciosum extract was carried out using thin layer chromatography (TLC) and nuclear magnetic resonance spectroscopy (NMR) techniques. The results indicated that the essential oil of T. ammi exhibited more effective larvicidal activity compared to the D. speciosum extract. Specifically, the essential oil demonstrated LC50 values of 18 ppm for Cx. quinquefasciatus and 19 ppm for Ae. aegypti. In contrast, the D. speciosum extract showed the strongest larvicidal effect against An. stephensi, with an LC50 of 517 ppm. Concentrations of 40 ppm of the essential oil and 1280 ppm of the extract resulted in 100% mortality across all three species. Both the essential oil of T. ammi and the D. speciosum extract exhibited concentration-dependent larvicidal activity, and these results were statistically significant (p < 0.001) compared to the no-treatment group. GC-MS analysis revealed thymol (88.95%), o-cymen-5-ol (4.11%), and γ-terpinene (2.10%) as the major constituents of the T. ammi essential oil. Additionally, TLC verified the presence of alkaloids in both chloroform and methanolic extracts. Proton NMR identified a diterpene structure for these alkaloids. These findings suggest that T. ammi essential oil is a promising candidate for natural mosquito control strategies. Given its efficacy, further research is warranted to explore its potential in integrated vector management programs.

HPTLC quantification of garcinol in three endemic Garcinia species of Assam, estimation of secondary metabolites and evaluation of in vitro antioxidant activity

AbstractGarcinol, the main polyisoprenylated benzophenone present in Garcinia species. A simple, rapid, reliable high-performance thin-layer chromatography (HPTLC) technique has been developed for simultaneous detection and quantification of garcinol in three endemic Garcinia sp. of Assam, India. While G. mangostana and G. indica have garnered extensive recognition, these lesser-known species have not received proportionate level of scientific exploration, therefore has served as the subject of investigation. Chromatographic separation was achieved using pre-coated silica gel 60 F254 plates. The optimized mobile phase (Toluene-Ethyl acetate-Formic acid, 5:4:1V/V) at 276 nm produced well-defined and sharp peaks of garcinol, maintaining a constant RF value of 0.75. The regression equation demonstrated a linear relationship with a correlation coefficient of 0.9955. The method was validated in accordance with International Conference of Harmonization guidelines (ICH), encompassing assessments for precision, accuracy, repeatability and robustness, all of which yielded low percentage of relative standard deviation values, attesting to its excellent performance. The content of garcinol found in the extracts were in the range of 0.58–0.98% w/w. In secondary metabolite analysis, significant amount of phenols (26.06 ± 1.65–57.51 ± 2.62 gallic acid equivalent mg g−1), flavonoids (11.27 ± 1.06–21.09 ± 1.34 quercetin equivalent mg g−1) and ascorbic acid (3.56 ± 0.12–3.65 ± 0.11 mg/10 g) contents were detected in the extracts. Further, the extracts exhibited good anti-radical activity, which may be attributed to the presence of important secondary metabolites. Therefore, this research establishes the three endemic Garcinia sp. as promising source of garcinol. Additionally, the developed method can be applied for routine quality control analysis of herbal formulations containing garcinol.

A Brief Review on Separation Techniques: Chromatography Techniques

Chromatography is a system that helps distinct, purify, and recognize mixture components for both quantitative as well as qualitative study [1]. Proteins can be separated and purified depends on their dimensions, shape, net charge, aquaphobic structures on the outermost layer, and binding capability with the immobile phase. The exchange of ions, superficial adsorption, separation, and size exclusion are examples of molecular property and interaction-based separation techniques. Stationary bed chromatography procedures, such as paper chromatography technique, Column Chromatography technique as well as Thin Layer Chromatography technique. Column chromatography technique serves as one of the most common protein purification methods. This review comprises an insight the separation techniques such as Column Chromatography technique, Gas Chromatography technique, Paper Chromatography technique, Thin Layer Chromatography technique, Hydrophobic Interaction Chromatography technique (HIC), used for separation of biomolecules.

Does Trema micranthum (L.) Blume Produce Cannabinoids?

There are inconclusive claims in the scientific literature that the species Trema micranthum, widely distributed throughout the Brazilian territory, may produce phytocannabinoids, potentially serving as an alternative to Cannabis sativa. In this study, we conducted a comprehensive investigation to assess the presence of phytocannabinoids in two Trema micranthum samples collected in the Midwest region of Brazil. In trying to detect cannabinoids in T. micranthum, a recommended cannabis screening test was employed, the Fast Blue BB Salt (FBBBS) colorimetric assay, followed by thin-layer chromatography (TLC) and instrumental techniques: high-performance liquid chromatography coupled to diode array detector (HPLC-DAD) and gas chromatography coupled to mass spectrometry (GC-MS). When employed without chloroform extraction, the FBBBS reagent yielded positive results for extracts from all parts of T. micranthum (leaves, branches, fruits, and inflorescences). However, these initial positive results from the FBBBS test, suggesting the presence of cannabinoids, were not corroborated by FBBBS followed by chloroform extraction, TLC, or the instrumental techniques used in this study. These additional outcomes suggest that the positive FBBBS test results were likely due to the presence of other phenolic compounds rather than phytocannabinoids. For example, the presence of vitexin-like compounds in T. micranthum extracts might explain the positive FBBBS test results. Therefore, new assertions that T. micranthum produces cannabinoids will require the support of more selective experiments to avoid false-positive claims based on less selective screening tests.

Impact of altitudinal variation on secondary metabolites, antioxidant, anti-inflammatory and anticancer potential of Adiantum venustum D. Don

Phytoconstituents are being emphasized as chemotherapeutic agents, as well as probable anticancer agents. The purpose of the present research was to assess the antioxidant, anti-inflammatory, and anticancer properties of crude ethanolic extracts and bioactive fractions of Adiantum venustum D. Don collected from three sites in Himachal Pradesh, India. The bioactive phytoconstituents were examined qualitatively and quantitatively in crude ethanolic extracts and fractions of all sites, and biological activities were analyzed. The potential presence of terpenoids, saponins, tannins, phenols, and flavonoids was examined, and it came to light that BF-II [n-butanol fraction of Site-II (2000 m altitude)] had a significantly higher phytochemical concentration. Additionally, the crude ethanolic extracts and fractions were examined for antioxidant and anti-inflammatory properties, and BF-II was found to have potential antioxidant and anti-inflammatory activity, as verified by the IC50 values (lowest half inhibitory concentration) in the DPPH (57.86 ± 0.72 μg/mL), reducing power assay (45.20 ± 0.72 μg/mL), FRAP (73.12 ± 0.23 μM Fe equivalent), BSA (56.76 ± 0.25 μg/mL), and EAA (EAA59.84 ± 0.13 μg/mL). Following that, the anticancer potential was investigated further using the MTT assay on the HeLa cell line, MTT assay revealed that BF-II possesses significantly higher anticancer potential. Furthermore, the dominant crude ethanolic extract and fraction were further investigated using gas chromatography-mass spectroscopy, and high-performance thin layer chromatography techniques from the selected plant. Overall, the study suggests that the n-butanol fraction of A. venustum possesses significantly higher therapeutic value but is still not explored to the extent of its potential.

Phytochemical analysis and the first report of decyl gallate and 4-ethoxycoumarin from marine seagrass Posidonia oceanica collected from Benghazi beach Libya

The goal of this research is to examine the chemical composition of the plant and identify the phytochemical constituents of the endemic seagrass species in the Mediterranean Sea, Posidonia oceanica, which has historically been used for a variety of purposes. After doing chemical screening, the existence of triterpenoid and phenolic proteins was discovered. Using preparative thin-layer chromatography and flash column chromatography techniques, seven phenolic compounds were isolated from the methanolic extracts of several plant parts. Namely decyl gallate, sinapinic acid, alpha-methyl cinnamic acid, acetosyringone, and chlorogenic acid from leaves, 4-ethoxycoumarin from rhizomes, and syringic acid from roots. The structures of these compounds were elucidated using spectroscopic methods such as ‘H and 13C-NMR. The spectra were analyzed and visualized using Mnova software and compared with standard spectra from the Willy spectra database, biological magnetic resonance data bank, and chemical book. This is the first report of decyl gallate and 4-ethoxycoumarin from the plant. This study contributes to the understanding of the phytochemical profile of Posidonia oceanica and provides valuable insights into its potential applications in various fields, including medicine, agriculture, and industry.

Development of Wound Gauze from Hylocereus undatus Dragon Fruit (Pigment) Peel Extract

The dragon fruit peels (Hylocereus undatus) are typically thrown away as agricultural waste. The disposal of bio-wastes, like peels from dragon fruit and microbiological contamination during their mass breakdown, all contribute to environmental degradation. Peel can be used to extract naturally occurring colour for pigments. The objective of this study is to investigate the feasibility of using the peel of Hylocereus undatus as a coating material for wound healing. The extraction method and potential uses of the pigments derived from the peel will be explored. The study aimed to determine the phytochemical components of Hylocereus undatus (peel) extract and the peel can be utilized as a raw material for pigment extraction due to the betalain content. With the use of a magnetic stirrer extraction, the peel of Hylocereus undatus was extracted with aqueous extraction. Thin layer chromatographic techniques are used to identify the pigment components. In the present study, we recorded both UV-VIS and FT-IR profile of peel extract to known the various phytoconstituents and determine the functional group present in Hylocereus undatus (white dragon fruit). The pigment needs to be applied to the wound gauze, and the stain resistance should be evaluated by determining how well the wound gauze can withstand contamination from the outside coating. Furthermore, the pharmaceutical and nutraceutical sectors are interested in Hylocereus undatus peel due to its components, which may have potential health applications such as anti-inflammatory, antioxidant, antibacterial, and anti-cancer characteristics

Synthesis and Preliminary Biological study of New Chalcone Derivatives Containing Isoxazoline Moiety

In this research work, synthesis, antimicrobial and antioxidant bioactivity of a chain of compounds having unsaturated ketones bond and isoxazoline moiety have been described. New chalcone derivatives containing isoxazoline moiety have been synthesized. Generally, Chalcones are unsaturated ketones bearing (-CO-CH=CH-) as reactive ketoethylenic group that give the bright yellow colored compounds due to this chromophore group. Firstly, chalcones (IIa-d) have been prepared by cyclocondensation (Claisen-Schmidt condensation) of triphenyl aminobenzaldehyde with different substituted acetophenone in ethyl alcohol to produce a series of chalcones compounds with bright yellow colored as a starting material,. The next step involving the novel chalcones (IIa-d) reacted with hydroxylamine hydrochloride afforded a new isoxazoline compounds (IIIa-d) in basic medium. The prepared compounds were fully characterized by melting points, FTIR, 1H-NMR spectroscopy, 13 C-NMR for some compounds and CHNS techniques. The reactions have been monitored by TLC (Thin Layer Chromatography) technique. The synthesised compound IId showed significant bioactivity against the gram-negative bacteria species. Also, the antioxidant activity of the some new synthesized compounds was evaluated and determined against the DPPH radical (1.1-diphenyl-2-picrylhydrazyl) and compared to that of a standard natural antioxidant, Ascorbic acid, compound IIb showed higher antioxidant activity by using the free radical DPPH. The outcomes of investigation enhanced the activity of new derivatives as antimicrobial reagent.

Marine Bacillus haynesii chitinase: Purification, characterization and antifungal potential for sustainable chitin bioconversion

The development of chitinase tailored for the bioconversion of chitin to chitin oligosaccharides has attracted significant attention due to its potential to alleviate environmental pollution associated with chemical conversion processes. In this present investigation, we purified extracellular chitinase derived from marine Bacillus haynesii to homogeneity and subsequently characterized it. The molecular weight of BhChi was approximately 35 kDa. BhChi displayed its peak catalytic activity at pH 6.0, with an optimal temperature of 37 °C. It exhibited stability across a pH range of 6.0–9.0. In addition, BhChi showed activation in the presence of Mn2+ with the improved activity of 105 U mL−1. Ca2+ and Fe2+ metal ions did not have any significant impact on enzyme activity. Under the optimized enzymatic conditions, there was a notable enhancement in catalytic activity on colloidal chitin with Km of 0.01 mg mL−1 and Vmax of 5.75 mmol min−1. Kcat and catalytic efficiency were measured at 1.91 s−1 and 191 mL mg−1 s−1, respectively. The product profiling of BhChi using thin layer chromatography and Mass spectrometric techniques hinted an exochitinase mode of action with chitobiose and N-Acetyl glucosamine as the products. This study represents the first report on an exochitinase from Bacillus haynesii. Furthermore, the chitinase showcased promising antifungal properties against key pathogens, Fusarium oxysporum and Penicillium chrysogenum, reinforcing its potential as a potent biocontrol agent.

Antimicrobial Activity of Fungal Endophytes Associated with Peperomia argyreia (Piperaceae)

The endophytic fungal biodiversity of unique plants like Peperomia argyreia (Miq.) É. Morren (Piperaceae) has antimicrobial properties and can be employed for infection treatment. Fungal isolates were obtained from appropriately treated plant tissues cultured in solid media, characterized by morphology, and identified by molecular biology using ITS and NL primers. The antimicrobial properties of fungal extracts were analyzed by combining microdilution and bioautographic assays complemented with metabolic profiling by automated thin-layer chromatography and 1H NMR techniques. Thirty-one filamentous fungi were isolated and characterized by ITS and/or D1/D2 region amplification of rDNA, identified as Thermothielavioides, Trichoderma, Cyphellophora, Cladosporium, Arcopilus, Plectosphaerella; Chaetomium, Sporothrix, Alboefibula, and Penicillium. Thermothielavioides spp. inhibited Staphylococcus aureus ATCC 25923; moreover, Penicillium westlingii P4 showed inhibitory activity on Ascochyta rabiei AR2. The bioactivity-guided fractionation of the EtOAc extract (MIC = 62.5 μg/mL) of P. westlingii P4 allowed the purification of citrinin as the main inhibitory compound (MIC = 62.5 μg/mL). Peperomia argyreia harbors a rich and diverse endophytic community able to produce bioactive molecules. Citrinin, with a minor influence of volatile compounds biosynthesized by P. westlingii P4, was responsible for the inhibition of A. rabiei AR2.

Amino acid profile and potential biomass conversion of Vitellaria paradoxa fruit pulp

Vitellaria species is one of Africa's most important fruit crops with enormous human benefiting nutrients including amino acids. This study characterised Vitellaria paradoxa shea nut pulp from 5 regional zones of Ghana based on its acidity, pH and among the few for its qualitative and quantitative amino acids profile. Thin Layer Chromatography (TLC) techniques, using cellulose TLC plates were used to profile and quantify the amino acids. Fifteen essential amino acids were detected in the Vitellaria paradoxa shea nut pulp. Alanine, arginine, glutamic acid, glutamine and histidine were common in the Vitellaria paradoxa shea nut pulp. The most concentrated (41.02 g/100 g) of the 7 essential amino acids in the shea nut pulp was arginine and the least was glycine, 2.12 g/100 g. All seven 7 essential amino acids, except leucine and valine, were significantly different (P < 0.05) across the 5 shea regional zones of Ghana. The most acidic pulp, with 18.63% acidity, was from the Upper West Region. The quality and quantitative of amino acids in Vitellaria paradoxa shea nut pulp differed significantly from one region to the other. The shea nut pulp is a good source of essential amino acids and can be used fruit juice production and provide the required N for an effective fermentation to proceed. This has remarkable economic implication and significant environmental effects. Some health benefits of eating shea nut fruits consequential of its nutritional effects due to its amino acids composition have been noted to include its ability to help improve mood, boost exercise performance, prevent muscle loss and promote weight loss. Moreover, the amino acids profile of shea nut pulp may provide researchers with useful applications of the Vitellaria paradoxa shea nut pulp in the various industries.

Synthesis, Identification, Biological Evaluation of Seven-membered Heterocyclic Derivatives from 2,3-di Chloroaniline

This study?s focus was centered on the synthesis of heterocyclic compounds incorporating a seven-membered ring, specifically 1,3-oxazepine. The initial phase of the synthesis involved the preparation of azo compound (1) through the coupling of the diazonium salt of 2,3-dichloroaniline with 4-bromoacetophenone in an alkaline alcoholic medium. This was then succeeded by the reaction of azo compound (1) with 4-nitroaniline and 4-methoxyaniline in absolute ethanol, employing glacial acetic acid as a catalyst, resulting in Schiff base derivatives (2) and (3), respectively. Subsequent to this, Schiff base (2) was reacted with phthalic anhydride and maleic anhydride, as well as succinic anhydride, in a dehydrated benzene environment, leading to the formation of heterocyclic seven-membered compounds (4), (5), and (6), respectively. Finally, Schiff base (3) was subjected to reaction with phthalic anhydride and maleic anhydride, as well as succinic anhydride, in dry benzene, resulting in the synthesis of heterocyclic seven-membered compounds (7), (8), and (9) respectively. Each of these synthesized compounds underwent characterization through Fourier-transform infrared and proton nuclear magnetic resonance spectroscopic analysis, and the progress of reactions was monitored through Rf and thin-layer chromatography techniques. Melting points were recorded as part of the characterization process. In addition, an investigation into the biological activity of these compounds was conducted against both positive and negative bacterial strains.. KEYWORDS :Azo compounds, Oxazepine, Schiff bases.

PREPARING AND CHARACTERIZING SOME NEW DERIVATIVES OF ORGANIC CYCLIC COMPOUNDS DERIVED FROM SOME DRUGS AND STUDYING THEIR BIOLOGICAL

This study involved synthesis some of derivatives via that a Suzuki-Miyaura cross-coupling procedure was used in order to produce a wide variety of Baclofen derivatives. After beginning with the reaction of 4-methoxy carbonyl phenyl boronic acid by the use of the reflex method, the treatment of Baclofen with a number of other boronic acids was carried out. The subsequent stage was the incorporation of Pd(0)(PPh3)4, and lastly, once the reaction had reached a state of equilibrium,Ethyl Acetoacetate ( EtOAc) was incorporated into the mixture. Taking this action was necessary in order to finish the procedure. Using the thin layer chromatography (TLC)technique, the observation of the reaction was carried out in order to carry out the observation. In order to accomplish the job of characterizing all of the unique Baclofen derivatives that were discussed in the preceding paragraphs [1-6], the analytical procedures of Fourier Transform Infrared (FT-IR) , Proton nuclear magnetic resonance (1HNMR) ,Carbon-13 nuclear magnetic resonance (13C-NMR) , and C.H.N were used. Furthermore, a wide range of other physical characteristics, such as color, Rf, and mechanical capabilities, were investigated and appraised. The objective of the experiment was to determine whether or not the treatment was effective against two different types of bacteria according to the biological standards. One of the objectives of the experiment was to do this. There was one species of fungus, which was Candida albicans, and one of the bacteria was positive (Staphylococcus aureus). The other bacterium was negative (Escherichia coli), and there was one type of bacteria. Of the bacteria, two were found to be positive. Following the assessment of the compounds' bacterial inhibitory biological activity [1-6], it was shown that the derivatives had an exceptionally high degree of effectiveness at a certain concentration. An additional accomplishment was the successful completion of a molecular docking investigation of novel Baclofen derivatives. [1-6].

Garlic Extract as a Bath Treatment against Gyroductylus sp. in Guppy Fish (Poecilia reticulata)

Parasitic infection is considered one of the major constraints in commercial fish farms owing to huge economic losses. Although there are plenty of commercial drugs available for treatments, those synthetic chemicals are more hazardous to the host and the environment than the parasite. Therefore, a special trend can be seen towards herbal medications. The present study is utmost significance as it focused on a well-known spice, Garlic (Allium sativum) as a bath treatment against gyrodactylosis caused by Gyrodactylus sp. The soxhlet extraction method was used to haul out the water-soluble compounds and the major components were identified using Thin-Layer Chromatography (TLC) techniques. During the phytochemical analysis, flavonoids, glycosides, quinones, and diterpenes were identified where the alkaloids, tannins, and phenols were absent in garlic extract. To find out the LC50, values acute toxicity test was performed in 72 hours with triplicates and 802.2ppm value was resulted. 216 of infected adult guppies (2.9 ± 0.1cm, 0.32 ± 0.05 g) were used for a three-hour in vitro bath treatment with different concentrations of garlic extract including 50, 100, 150, 200, and 250 mg/L. Complete recovery from gyrodactylosis is observed by the means of microscopic observations at 3, 2.5, 2, and 1.5 h with 50, 100, 150, and 200mg/L of concentrations, respectively. According to the observations, it can be concluded that exposure to garlic extract caused the parasite to first detach from the host, then restrict its movements, and finally die. A significant negative correlation (p&lt;0.01) can be observed between the garlic concentration and the eradication time. 250 ppm garlic extract bath for 0.5 hours was identified as the optimal and most effective treatment against gyrodactylosis. Therefore, garlic extract can be suggested as a herbal bath treatment against agent Gyrodactylosis infection in guppy fish. &#x0D; Keywords: Bath treatment, LC50, Gyrodactylus sp, Poecilia reticulata

A comparative investigation of bioactive compounds present in Datura stramonium L. leaves extract in various solvents (ethanol, aqua, and ether) using TLC, IR, and GC-MS analytical techniques

The present study is an experimental comparative investigation of the biomolecules present in ethanol, aqua and ether extract of Datura stramonium L. leaves. The purpose of the study is to monitor the phytochemical constituents in the plant leaves by thin layer chromatography (TLC), Gas Chromatography-Mass Spectrometry (GC-MS), and infrared spectroscopy analytical techniques. The Datura Stramonium L. leaves were collected, dried and was extracted with ethanol, water and ether at room temperature. A comparative investigation of bioactive compounds have been carried out using TLC, GC-MS, and IR Analytical techniques. Preliminary phytochemical analysis revealed the presence of protein, alkaloid, phenol and tannins, flavanoid, glycoside, sterols, saponin, resins, anthocyanin, and carboxylic acid. More number of bio active phytochemicals were identified, and the chromatograph showed peaks of individual compounds. The result of this study offers a platform for understanding the Phytochemical present in ethanol, aqua and ether extract of studied plant leaves and to identify them as herbal alternative for various diseases and it can be used as functional pharmaceutical when once it extracted and treated carefully.