Thin-layer Chromatographic Technique Research Articles

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.

Influence of various cooking treatments and extraction solvents on bioactive compounds and antioxidant capacities of bottle gourd (Lagenaria siceraria) fruit in India

Bottle gourd is a good source of health-beneficial bioactive substances. This research aimed at evaluating the effect of different cooking methods (pressure cooking, microwaving, and frying) and extraction solvents (methanol, ethanol and butanol) on bottle gourd fruit phenolic compounds and antioxidant capacity. The quantitative estimation of polyphenolic compounds, flavonoids and tannins was estimated by spectrophotometric methods. The antioxidative properties were evaluated using ferric thiocyanate (FTC), thiobarbituric acid (TBA), ferric reducing antioxidant power (FRAP) and 2,2-diphenyl-1-picrylhydrazyl (DPPH) tests using the standard methods. Moreover, quantification of individual polyphenols was carried out by high-performance thin layer chromatography (HPTLC) technique. Frying and pressure-cooking thermal treatments were observed to be the best methods for retaining bioactive molecules. When compared to the raw counterpart, the retention level of total phenol content (TPC) in pressure cooked and fried samples was 23.8% and 13.3%, respectively. Similarly, antioxidant capacity in terms of FRAP was observed to increase by 47.26% after pressure cooking and 28.50% after microwave cooking in respect to the raw sample. The HPTLC results showed that this fruit has high antioxidant capacity and is rich in phenolic acid [gallic(17.83 mg g−1dwb), p-coumaric (6.70 mg g−1dwb)and vanillic (60.66 mg g−1dwb] and flavonoids [quercetin (24.64 mg g−1dwb) and myrecetin (20.73 mg g−1dwb].The chromatography indicated destruction and transformation of new phenolic compounds due to heat treatments. The correlation analysis revealed that flavonoids were much more responsible for their antioxidant activity. Cooking methods and extraction solvents affected the retention and recovery of polyphenolic compounds available in bottle gourd fruit. These findings offer valuable information for optimizing cooking techniques to preserve and enhance the nutritional and antioxidant properties of bottle gourd, making it a valuable fruit for a healthy diet. Pressure cooking could be the most suitable cooking treatment as far as retention of bioactive compounds like phenolic acids is concerned.Graphical

A Review on High Performance Thin Layer Chromatography

A common excellent replacement for GC and HPLC is High-Performance Thin Layer Chromatography (HPTLC), an improved and automated technique of thin-layer chromatography (TLC) that provides better separation performance and detection limits. Applications of HPTLC include the study of biological materials and phytochemicals, the measurement of herbal medications and active components, formulation fingerprinting, and the identification of adulterants in formulations. Using HPTLC, chemicals of forensic importance can be located. It is more sensitive and feasible to run many samples in a little period of time by using a small volume of solvent. It is one of the more intricate instrumental procedures, utilizing every feature available in thin-layer chromatography.

Redox-active green-electrode: Plant-extract based rutin and caffeic acid functionalized MWCNT for scanning electrochemical microscopy imaging and selective electrocatalytic oxidative sensing of dopamine

A straightforward, environmentally friendly, and rapid in-situ electrochemical functionalization of Rutin (Rut) and Caffeic acid (Caf) on a multiwalled carbon nanotube-modified glassy carbon electrode, denoted as GCE/MWCNT@Herb-Redox (Herb-Redox =redox active species from the plant extract), has been demonstrated. This method utilizes an eco-friendly, plant-based water extract from Indian Borage (Coleus amboinicus), referred to as IBE, as a precursor sample. The as-prepared molecular bio-electrode showed a well-defined and stable surface-confined redox response at standard electrode potentials, Eo’ = 0.160 and 0.200 V vs Ag/AgCl in pH 7 phosphate buffer solution. By performing physicochemical and molecular characterisations using TEM, SEM, UV–Vis, FTIR, high-performance thin-layer chromatographic techniques, it has been confirmed that Rut and Caf from the Herbal-plant were selectively immobilized on the MWCNT-modified electrode. Using scanning electrochemical microscopy (SECM), the electroactive sites of the molecular-electrode were mapped. The electrocatalytic function of the GCE/MWCNT@Herb-Redox was demonstrated by performing a dopamine (DA) oxidation reaction as a model system in pH 7 PBS using CV and amp-it techniques. The results showed 100—10,000 times increase in the DA- oxidation current response over the values quoted with many of the literature reports highlighting the efficient functionality of the new molecular electrocatalytic system. As a proof of concept, batch injection analysis based elegant dopamine sensing using screen-printed electrode/MWCNT@Herb-Redox as an electrochemical detector has been demonstrated and there is no biochemical interference was observed. Since the electrochemical approach follows a green chemistry route for the modification, this bioelectrode showed a new platform for eco-friendly electrocatalytic applications.

Phytochemical stimulants for cancer therapeutics from Garcinia gummi-gutta: A prime research report

Cancers and tumors grow day by day and alternate traditional-based medicines are in demand to nullify the toxicity and side effects caused by modern pharmacological compounds. The medicinal properties of plant-based Indian ethnic drugs make them extremely important in this category. Traditional uses of G. gummi-gutta include a variety of herbal and folk remedies. Different herbal parts are extracted, isolated their specific compounds are targeted for applications in modern medicinal research today. The fresh fruit of G. gummi-gutta was extracted by ethyl acetate and tested through phyto-chemical modes. Thin layer Chromatography (TLC) and Gas Chromatography Mass-Spectrometry (GC–MS) were used to purify and characterize bioactive compounds. Studies using antimicrobial agents were conducted on gram-positive and gram-negative bacteria, including Salmonella typhi, Pseudomonas aeruginosa, Escherichia coli, Staphylococcus aureus, and Streptococcus mutans. The MCF-7 breast cancer cell line was used to examine cellular cytotoxic activity. Software for BSP-SLIM molecular docking analysis was utilized together with in-silico virtual screening techniques. The receptor structure was retrieved from the protein databank (Structure of human Bcl-2) in a complex with an analog of ABT-737 (6QGG). Pymol was found to have the best ligand interactions. The protein's stability characteristics were confirmed and molecular dynamics were estimated using a programme called Dynomics. A preliminary qualitative phytochemical investigation revealed that the primary constituents of G. gummi-gutta include alkaloids, saponins, tannins, phenol, flavonoids, glycosides, and protein. The compound positions and Rf values were disclosed by the TLC technique. GC–MS technique revealed the presence of pure compounds whereas the antimicrobial activity was appraised through the disc diffusion method and the relative zone of inhibition was determined. The MTT test was used to measure the antiproliferative activity against the MCF-7 breast cancer cell line at a dose of 7.8 μg/mL. The virtual screening using the in-silico method further revealed the protein-ligand interactions and molecular dynamics that assisted in identifying the protein functions and stability graphs. G. gummi-gutta has thus proven to be an anticancer medicinal herb both by in-vitro and in-silico studies. This could further be applied for drug designing and database creation leading to machine-based diagnostics and therapeutics.

Determination of Aflatoxins in Pig Feed. Using the Thin Layer Chromatography Technique

Aflatoxins are the most important mycotoxins in terms of food safety, due to their frequency and toxicity, they are characterized by being secondary metabolites synthesized by fungi. In order to determine the presence of aflatoxins in pork feed, five samples from different commercial companies were analyzed. The methodology used prior to the sample extraction process was the thin layer chromatography technique, observed using long-wave ultraviolet light. It could be seen in samples 1,3,4,5, the presence of Aflatoxins, toxicogenic strains of Aspergillus parasiticus, which produce aflatoxins, G1 and G2, is characterized by the fluorescence coloration, in sample 2 the presence was observed of aflatoxins toxicogenic strains of Aspergillus flavus. Toxigenic strains of Aspergillus flavus generally produce only aflatoxins B1 and B2, while toxicogenic strains of Aspergillus parasiticus produce aflatoxins B1, B2, G1 and G2. AFB1 aflatoxins have been considered obvious carcinogens in experimental animals. It is important to carry out systematic research with low-cost analytical techniques to understand the mechanisms of production and action of aflatoxins in foods, in order to prevent, monitor and control these types of toxins.

Structure elucidation of active compounds from Coffea canephora Pierre ex A.Froehner cascara and their potential as anticancer against breast cancer cells

Context: One approach to cancer therapy medication is exploring medicinal plants that contain one or more compounds specifically targeting cancer cells with fewer side effects. Cascara from coffee fruit (Coffea canephora Pierre ex A.Froehner) is a waste rarely processed but has various chemical contents that can be used in medicine. Aims: To evaluate the in silico and in vitro activity of compounds isolated from ethanolic extract of C. canephora cascara against HeLa and MCF-7 breast cancer cells. Methods: Isolation of the compounds by radial chromatography and thin layer chromatography techniques, and the chemical structures were elucidated by infrared radiation, ultraviolet, nuclear magnetic resonance spectroscopy, and mass spectrometry. In silico study about the compounds binding with the receptor responsibility to cancer (caspases 3 and 9). In vitro study by examining the cytotoxicity of HeLa and MCF-7 cells of the isolated compounds from C. canephora. Results: Four known bioactive compounds, lupeol (1), stigmasterol (2), ursolic acid (3), and caffeic acid (4), were isolated from the ethanol extract of C. canephora cascara. Based on the ESI-MS results, the m/z value for lupeol was 427.50 [M+H]+, stigmasterol was 454.48 [M+ACN+H]+, ursolic acid was 456.51 [M+H]+, and caffeic acid was 179 [M-H]. In silico and in vitro data show that the ursolic acid compound has activity against HeLa and MCF-7 cancer cells with IC50 values of 25.98 ± 0.01 µg/mL and 12.70 ± 0.11 µg/mL, respectively. Conclusions: All isolated compounds from C. canephora cascara have a promising ability to interact with caspases 3 and 9, particularly ursolic acid, which has the smallest IC50 value against HeLa and MCF-7 breast cancer cells.

Evaluation of Lipids for the Study of Photosynthetic Membranes.

The biological role of lipids goes far beyond the formation of a structural membrane bilayer platform for membrane proteins and controlling fluxes across the membranes. For example, in photosynthetic thylakoid membranes, lipids occupy well-defined binding niches within protein complexes and determine the structural organization of membrane proteins and their function by controlling generic physicochemical membrane properties. In this chapter, two-dimensional thin-layer chromatography (2D TLC) and gas chromatography (GC) techniques are presented for quantitative analysis of lipid classes and fatty acids in thylakoid membranes. In addition, lipid extraction methods from isolated thylakoid membranes and leaves are described together with a procedure for the derivatization of fatty acids to fatty acid methyl esters (FAME) that is required for GC analysis.

Secondary metabolites from the leaves of Terminalia myriocarpa and their α-glucosidase inhibitory potential

The objective of this study is to investigate the chemical composition of the leaves of Terminalia myriocarpa and to evaluate their α-glucosidase inhibitory activity aiming to be used as a safe antidiabetic. Consequently, 10 compounds were isolated using column and preparative thin-layer chromatographic techniques and identified as alphitolic acid, isovitexin, flavogallonic acid, nigaichigoside F1, quercetin, quadranoside IV, rosamultin, 19α-hydroxyasiatic acid, asiatic acid, and arjunic acid. Structure elucidation was based on chemical (acid hydrolysis) and spectroscopic (UV, 1 H, and 13C-NMR) analyses and findings were confirmed through comparison with published data. The α-glucosidase inhibitory activity of the methanol extract fractions and compounds isolated therefrom was evaluated in-vitro alongside that of acarbose (positive control). Results revealed that quercetin and flavogallonic acid were significantly active with half-maximal inhibitory concentrations (IC50) equal to 7.5 ± 0.09 and 21.0 ± 1.4 µM, respectively. Furthermore, the interaction of lavogallonic acid with α-glucosidase enzyme was verified by docking experiment. Accordingly, quercetin and flavogallonic acid could be considered safer natural alternatives to the currently availableα-glucosidase inhibitors.

Isolation, Characterisation and Corrosion Inhibition Evaluation of 1, 3, 5-Trimethoxymethylbenzenefrom Seeds Extract of Cola nitida on Mild Steel Surface in Hyhrochloric Acid Solution

Corrosion inhibition of Mild steel in 1 M hydrochloric acid solution by 1, 3, 5-trimethoxymethylbenzene (TMMB) isolated from seeds extract of Cola nitida was studied using weight loss, potentiodynamic polarization and electrochemical impedance spectroscopic methods. TMMB was isolated from seeds of C. nitida by 72 hours maceration of the chopped seeds with dichloromethane. Filtered and concentrated extract was subjected to a combination of vacuum liquid, column and thin layer chromatographic techniques. The isolated compound was characterized using Fourier Transform Infra-Red (FT-IR) spectrophotometer, proton nuclear magnetic resonance (+H-NMR), C13 – NMR and DEPT-135 NMR spectrophotmeters. The results showed that the inhibition efficiency recorded by the weight loss method increased with the increase in TMMB concentration, but decreased with increasing temperature. The inhibition efficiencies recorded by the potentiodynamic polarization and electrochemical impedance spectroscopic methods also increased with increase in the concentration of TMMB. The calculated thermodynamic parameters showed that the corrosion inhibition process was endothermic and spontaneous in nature. Potentiodynamic polarization measurements confirmed that TMMB worked as a mixed – type inhibitor considering the value of Ecorrbeing less than 85 mV. Impedance measurement revealed that the charge transfer resistance offered by TMMB to the migration of aggressive ions on the surface of mild steel in acidic solution was enough to impede corrosion. The best line of fit to the adsorption of TMMB onto mild steel surface was found in Temkin isotherm. Formation of protective dense film on the steel surface in the presence of TMMB compared to the blank was revealed by scanning electron microscopy. The molecular structure of TMMB was optimized using Gausian 09 with density functional theory [DFT/B3LYP/6-31G (d,p)]. Physical adsorption has been proposed for the adsorption of TMMB onto mild steel surface.

Comprehensive high-performance thin-layer chromatography analysis of Monteverdia ilicifolia leaf and its adulterants

The leaves of Monteverdia ilicifolia (syn. Maytenus ilicifolia) are widely used in traditional South American medicine to treat gastrointestinal problems such as gastritis and ulcers. Several herbal products containing the leaves of M. ilicifolia can be found in the market. However, other species with similar leaf morphology are confounding materials, e.g. Monteverdia aquifolia (Celastraceae), Citronella gongonha (Cardiopteridaceae), Jodina rhombifolia (Santalaceae), Sorocea bonplandii (Moraceae) and Zollernia ilicifolia (Fabaceae). This study aimed to identify M. ilicifolia and distinguish it from its potential adulterants using high-performance thin-layer chromatography (HPTLC) technique. Comprehensive HPTLC analysis revealed specific fingerprints that can be used to assess the minimum content of epicatechin and the quality of commercial espinheira-santa samples. The results of the study demonstrated that the HPTLC method is capable of detecting adulterations and distinguishing M. ilicifolia from all confounding materials in commercial products available on the market, showing that most of the products are of poor quality due to adulterations.

Simultaneous Preconcentration of Fast Green FCF and Rhodamine B Using Deep Eutectic Solvent and Determination via High-Performance Thin Layer Chromatography

The aim of the current investigation is the development of a green, quick, easy, and accurate method for simultaneous preconcentration of fast green FCF (FG) and rhodamine B (Rh B) using a deep eutectic solvent (DES). Then, the high-performance thin layer chromatography (HPTLC) technique was used for the determination of analytes. Decanoic acid and tetrabutylammonium bromide were chosen as the components of DES. HPTLC analysis was performed on an aluminum plate silica gel 60 F254. Methanol-ammonia and ethyl acetate were selected as the mobile phase. Scanning of the plates was accomplished by scanner 3. Effective parameters on the preconcentration process such as concentration of salt, volume of DES, stirrer time, and pH were investigated via central composite design (CCD). Data validation demonstrated good repeatability. The limit of detection for FG and Rh B was obtained as 0.08 and 0.01 µg·mL−1, respectively. The enrichment factor for FG and Rh B was achieved as 7.43 and 10.77, respectively. The linear ranges for FG and Rh B were acquired as 0.10–1.20 and 0.05–1.20 µg·mL−1, respectively. The preconcentration factor for both analytes was 21.66. Finally, the proposed method was successfully used for the quantitation of FG and Rh B in pastille and lipstick.

Effect-Directed Assays and Biological Detection Approaches Coupled with Thin-Layer Chromatography as an Evolving Hyphenated Technique: A Comprehensive Review.

Bioautography is a technique for the detection of biological activity that combines the elements of planar chromatography. Its hyphenated variants are widely used in the screening of natural products possessing biological activity. It can be used in the activity-based screening of phytochemical ingredients by employing various enzyme processes and reactions and facilitates the rapid determination of bioactive compounds in pant samples. To give a comprehensive overview of effect-directed assays and biological detection approaches used in conjugation with thin layer chromatography technique. The present review article attempts to throw light on the various aspects of bioautography, including its types and applications, thereby giving its concise overview and its relevance in the field of natural product screening. Various search engines were used for the literature survey, including Google Scholar, Semantic Scholar, PubMed, ResearchGate and Scopus. Bioautography has wide-ranging uses in the screening of compounds such as antioxidants, antifungals, antimicrobials, estrogenic, antitumors, and various enzyme inhibitors compounds like α and β-glucosidase inhibitors and α-amylase inhibitors. Bioautography serves to be an effective tool for the isolation of bioactive phytochemicals, thereby allowing us to scientifically validate the biological activities of various compounds, which can then be utilized for making potent medications for various diseases.

Impressive merger between green analytical approaches and quality- by- design for alcaftadine determination in eye drops and rabbit aqueous humor; application to stability study by two validated chromatographic methods

Nowadays, the main target of many researchers is to develop green environmentally friendly techniques that could separate drugs without using hazardous solvents which may lead to a dreadful impact on the environment. Proceeding from this lofty goal, the main concern of this work is to develop two green chromatographic techniques, namely, HPLC and TLC to determine alcaftadine (ALF) in pharmaceutical formulation, rabbit aqueous humor, and in presence of its degradate. Fortunately, and for the first time in the history of ALF analysis, the developed HPLC technique in this work succeeded to determine ALF in the presence of its oxidative degradate via a fruitful integration between green analytical chemistry and analytical quality-by-design resulting in increasing efficiency and minimizing environmental impacts. Utilizing an XBridge® C18 (4.6 × 250 mm, 5 µm) column, optimized chromatographic separation was accomplished utilizing isocratic elution with an ammonium acetate buffer (pH 5.0): ethanol (60:40, by volume) as the mobile phase, a flow rate of 1.5 mL/min, and UV detection at 282.0 nm. Over a concentration range of 0.1 to 8.0 µg/mL, linearity was attained. Additionally, this work succeeded to determine ALF through a stability indicating study using thin layer chromatographic technique on TLC plates precoated with silica gel 60F-254 as a stationary phase using a developing system consisting of dichloromethane: ethanol: ammonia (5:5:0.1, by volume) and scanned at 282.0 nm with 0.5–20.0 µg/band linearity range. Finally, the proposed methods were further assessed for their greenness profile using various greenness assessment tools. Fortunately, the developed methods proved to be safe, green, and eco-friendly.

Quality by design optimization and validation of a HPTLC-MS method for simultaneous estimation of paracetamol and prochlorperazine from bulk and formulation

Study reports that the, stability indicating high-performance thin layer chromatographic technique (HPTLC) was developed as well as validated for estimating Paracetamol and Prochlorperazine in bulk and dosage form. These drugs are largely used in migraine and first line treatment of schizophrenia. Selected drug combination employed for Quality by design based Box Behnken design by high performance thin layer chromatography method utilizing aluminium plates; pre-coated using silica gel 60F254 as stationary phase by using acetone: toluene: methanol (4:4:2 v/v/v) as a mobile phase. Densitogram shows Rf values for Paracetamol at 0.74 and for Prochlorperazine at 0.32 evaluated by densitometry measurement bands at 248 nm. Paracetamol and Prochlorperazine marketed formulation of 700 mg by using various excipients prepared at lab scale and it was evaluated for various types of tablet tests, and it showed the accurate result with respect to standard guidelines. The approach has been validated in accordance with International Conference on Harmonization (ICH) requirements. The correlation coefficient was found to be 0.992 and 0.998 at a concentration range of 100–300 μg/band and 1000–3000 ng/band for Paracetamol and Prochlorperazine, respectively. The method had an accuracy of 100.51 % and 100.53 % for Paracetamol and Prochlorperazine, respectively. The degradation of Paracetamol and Prochlorperazine was carried out in an acid, alkaline, oxidative, thermal and photolytic environment. The stress degraded product was successfully separated using the HPTLC and using high resonance mass spectroscopy technique (HRMS-MS), the degradation product was discovered.