Thin-layer Chromatographic Technique Research Articles

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.

In Vitro Trypanocidal Effect of the Extract and Isolated Compound of Corymbia torelliana Stem Bark against Trypanosoma brucei brucei (Federe Strain)

African animal trypanosomosis (AAT) is a severe debilitating protozoan disease of livestock and some wild animal species. It is caused by the pathogenic trypanosome species, and transmitted by Tsetse flies. It affects about 60 million cattle in Africa, with about US$1.5 billion in annual losses. Eradication of the disease through chemotherapy and vaccine development has been hindered by parasite antigenic variation and its resistance to drugs. In this study, the crude ethanol stem bark extract of Corymbia torelliana was prepared by cold maceration in 70% Ethanol. It was further purified using the modified Kupchan partition method into n-hexane, Ethyl acetate, N-butanol and Aqueous pure fractions. A pure compound was isolated from the ethyl acetate purified fraction using the column and thin layer chromatographic techniques. The isolated compound was named Makoshi_Mak1. The in vitro antitrypanosomal effects of the crude extract, the purified fractions and the isolated compound were then tested against the blood stream forms of Trypanosoma brucei brucei (Federe strain). 0.2 ml of prepared trypanosome suspension containing about 2x104 parasites was dispensed into each of seven clean test tubes using micropipettes. 0.2ml of prepared Ringer’s solution was also added to stabilize the parasites. Five varying concentrations of the crude extract, the extract fractions and the isolated compound (Makoshi_Mak1) were added to test tubes 1-5. Test tube 6 and 7 served as the negative and positive controls respectively. Aliquots of the contents of each test tube were examined under the microscope at x400 magnification for parasites motility at different time interval for 2 hours. The crude extract was effective at the concentrations of 12 mg/ml, 1.2 mg/ml,0.12 mg/ml and 0.012 mg/ml. the n-hexane fraction had efficacy at 0.12mg/ml and 0.096mg/ml. The Eethyl acetate fractions were effective at 0.12mg/ml, 0.096 mg/ml, 0.048mg/ml and 0.024mg/ml, the N-butanol fraction had efficacy at 0.12mg/ml and 0.096mg/ml, while the Aqueous fraction was effective at 0.2mg/ml, 0.16mg/ml and 0.08mg/ml. Makoshi_Mak1 was effective in vitro against T. brucei beucei at all the tested concentrations of 0.219mg/ml, 0.109mg/ml, 0.055mg/ml and 0.027mg/ml. Efficacy of the extracts and isolated compound was determined by parasite mortality, and was seen to be concentration dependent. Dead parasites appeared to float freely on the fluid environment, while the lysed parasites were seen as floating debris. The results of the study showed that both the crude extract, the purified extract and the pure isolated compound of Corymbia torelliana had in vitro activity against Trypanosoma brucei brucei in a concentration dependent manner. Corymbia torelliana can then be considered as a potential herbal antitrypanosomal drug candidate.

Comparative Chemical Profiling of a Polyherbal Formulation with Respect to its Ingredient Plants Using Spectrophotometric and High-performance Thin-layer Chromatographic Techniques

Background Guggulutiktam Kashayam (GTK) is a water decoction used in Ayurveda for the treatment of various diseases like skin diseases, ulcers, anemia, thyroid problems, stress, liver disease, and leucorrhoea. It is a polyherbal formulation prepared from specific parts of 28 medicinal plants. Objectives The study aimed at a comparative phytochemical evaluation of an important polyherbal formulation with its ingredient plants. Materials and Methods Chemical profiling of GTK has been carried out in comparison with its ingredient plants. Spectrophotometric estimation of major class compounds and high-performance thin-layer chromatography (HPTLC) profiling were done using a standardized protocol. Results The results of the study showed that the phytochemical contents of the finished formulation are different from those of the ingredient plants, which might be due to the numerous synergistic and antagonistic interactions of numerous chemical components extracted from different ingredient drugs. HPTLC-based comparative chromatograms showed the differences and similarities in certain compounds that are specific to some of the plant ingredients. Conclusion The study established the cumulative chemical interactions of various phytochemicals during the preparation of an important polyherbal formula used in Ayurveda.

Production, Purification, Characterization and Anti biofilm activity of natural antibiotic by Actinokineospora cibodasensis from marine water

Health sector faces a distress globally because of the scarcity of newer antibiotics against drug resistant pathogens. Innovations in developing new antibiotics is the solution to combat the drug resistant pathogens. This alarming situation requires advanced, potential and effective antibiotics. Secondary metabolite products from microbes remain as the main origin for many of the antibiotics in the market today and are also found to be promising till date. Out of the microbes, the major share of antibiotic production is reported by Actinomycetes, which are Gram positive, anaerobic, filamentous bacteria. They are currently being exploited through new approaches to improve the efficiency of commercially valuable antibiotics. Since natural environment stands a potential source of innovative and novel antimicrobials, this investigation was performed to test the potential of selected marine actinomycetes. Of the screened ones, Actinokineospora is potential enough to cease the growth of biofilm formation. This strain was identified by molecular characterization as Actinokineospora cibodasensis. It was selected to extract secondary metabolites by submerged fermentation and made to undergo further purification and structural analyses for the search of bioactive components. Presence of streptomycin like compound by Gas Chromatography Mass Spectrometry led to a breakthrough in the study which had been confined by High Performance Thin Layer Chromatography and Fourier Transform Infra-Red Spectroscopy techniques by comparing with standards. Gene expression studies showed that the selected strain can down regulate the genes in the formation of biofilm by the test organism Pseudomonas aeruginosa. The bioactive compound from Actinokineospora is an ecofriendly product along with a biofilm mitigating activity on medical devices. Rare actinomycetal isolate was able to produce the antibiotic which could apply on medical device to reduce the infections during the treatment. This antibiotic coating on life saving devices will provide the ultimate protection for patients.

A review on different types of detectors used in chromatography techniques

High Performance Thin layer Chromatography (HPTLC) technique is a sophisticated and automated form of the thin-layer chromatography (TLC) with better and advanced separation efficiency and detection limits. The analytical method should be sensitive, specific, fast and accurate to establish the assurance that the equipments used in manufacturing are free of the unwanted impurity, presence of which may alter the safety and efficacy of the drug product. The developed method was based on RP-HPLC leave-taking and quantification of the drug on C-18 column using a mobile phase at flow rate of 1 ml/ min. Quantitation was attained with PDA detector at 200-400 nm based on peak area with linear calibration curves at concentration ranges 5-25 μg/ml for the drug. The method as per ICH guidelines was validated for specificity, linearity, detection limit, quantitation limit, precision, accuracy, robustness, solution stability, and can be effectively used for routine analysis.

A Review of various Analytical Methods Developed for Estimation of Paracetamol, Diclofenac sodium, and Chlorzoxazone in Single-component and Multi-component Dosage Form

Painful muco-skeletal joint disorders such as osteoarthritis, rheumatoid arthritis, and ankylosing spondylitis are typically treated with a combination of paracetamol, diclofenac sodium, and chlorzoxazone. Paracetamol and diclofenac sodium are non-steroidal anti-inflammatory drugs, while chlorzoxazone is a muscle relaxant. It acts on the central nervous system and the spinal cord to reduce muscle stiffness and spasms as well as increase muscle mobility. The most commonly used pharmaceutical medicine is paracetamol, which comes in a variety of dosage forms such as injection, tablet, capsule, drops, elixirs, suspensions, and suppositories. However, due to toxicity, it may cause mortality in large dosages with extended intake. As a result, more efficient analytical approaches aimed at providing quality control of routinely used medications are required. Numerous analytical methods, both spectroscopic and chromatographic such as high-performance liquid chromatography, gas chromatography with mass spectrometry, and high-performance thin-layer chromatography have been established to estimate paracetamol diclofenac sodium and chlorzoxazone in their single-component dosage form. The simultaneous estimation of paracetamol, diclofenac sodium, and chlorzoxazone in combined dosage forms has been performed using reverse-phase high-performance liquid chromatography and high-performance thin-layer chromatography techniques. In contrast, there are very few techniques available to estimate paracetamol, diclofenac sodium, and chlorzoxazone in combination pharmaceutical dose form till date. This article is an attempt to jut out all the methods developed to estimate the above-mentioned drugs in the single or multi-component dosage form.