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INTRODUCTION. The determination of impurities is a key requirement for the quality assessment of medicines because impurities can significantly impact the quality and therapeutic effectiveness. Pharmacopoeias are the most important scientific and methodological guidelines for manufacturers developing medicinal product specifications and regulators assessing these specifications as part of registration dossiers. Therefore, it is essential to harmonise national and international approaches to impurities.AIM. This study aimed to analyse and summarise pharmacopoeial requirements for and methodological approaches to the control, evaluation, and identification of organic impurities in medicines.DISCUSSION. The authors compared requirements for the control of organic impurities in small-molecule medicines set forth in national and international pharmacopoeias and guidelines of the International Council for Harmonisation of Technical Requirements for Medicinal Products for Medical Use (ICH) and the Eurasian Economic Union (EAEU). This comparison highlighted the differences in current approaches that require further harmonisation of the existing regulatory documentation. Additionally, this study analysed the popularity, advantages, and disadvantages of different options for determining impurities in two-component combination products (i.e. identified and/or unidentified impurities in one or each of the active substances). The analysis demonstrated the need to control nitrosamines and genotoxic impurities and to use selective and highly sensitive chromatographic methods.CONCLUSIONS. When drafting pharmacopoeial monographs for medicinal products, experts should consider the general approach set forth in the EAEU pharmacopoeia and regulations and in the ICH Q3B guideline. This approach recommends that process-related impurities of active substances should not be controlled at the medicinal product level. Therefore, pharmacopoeial monographs for active substances should distinguish degradation products from process-related impurities. Impurities should be determined for each active substance to ensure the quality and safety of fixed combination medicinal products. Priorities for improving the methodological approach to the control of organic impurities include using reference standards for impurities and acknowledging the necessity of impurity quantification.

Introduction: Kefir beverage has beneficial microorganisms that have healthgiving properties; therefore, they have a good potential to be probiotic. This study evaluated the probiotic potential, technological, and safety characteristics of Enterococcus faecalis, Lactococcus lactis, and Pichia fermentans isolated from traditional kefir beverages. Method: First, isolates were evaluated in terms of resistance to acid, alkali, bile salts, trypsin, and pepsin of the gastrointestinal tract. The auto-aggregation and co-aggregation ability of isolates were measured using spectrophotometry. Antimicrobial activities were assayed against important food-borne pathogens using the agar well diffusion method. Moreover, gamma-aminobutyric acid (GABA) production was investigated by thin-layer chromatography (TLC). Result: Among the isolates, P. fermentans had an 85% total survival rate, but its amount reached below 6 log CFU/ml which is considered non-resistant, and it showed the highest auto-aggregation (74.67%). Moreover, only L. lactis showed antimicrobial activity and had the highest co-aggregation with E. coli PTCC 1338 (54.33%) and L. monocytogenes ATCC 7644 (78%). Finally, an evaluation of the technological and safety characteristics of the strains showed that the strains produced GABA and were safe. Discussion: Although the isolates were not resistant to the gastrointestinal tract, their supernatant contained valuable natural compounds, including antioxidants, GABA, and antimicrobials, which can be used to produce functional foods and medicines. In addition, other approaches, such as increasing the initial number of strains, using foods as carriers of isolates, and encapsulating the isolates, can effectively increase the survivability of isolates in the gastrointestinal tract. [ABSTRACT FROM AUTHOR]

Mycophenolic acid is a secondary metabolite of Penicillium brevicompactum and has been extensively studied for its antineoplastic, immunosuppressive, anti-inflammatory, antiviral, antipsoriasis, and antifungal activities. In the present study, a high-performance thin-layer chromatography method was developed for determining mycophenolic acid and a chitosan-linked prodrug using 60 F254 silica gel plates. The mobile phase used was chloroform–methanol–water (9.2 : 0.7 : 0.1, v/v/v), which provided good separation. The linearity was found to be in the range of 4–12 µg/spot with an R2 value of 0.9973. The percentage of RSD for intra-day and inter-day precision was found to be in the range of 0.7–1.1 and 0.9–2.0, respectively. The limit of detection and limit of quantification of the prodrug were determined to be 0.27 and 0.82 µg/spot, respectively. The recovery of the prodrug obtained was found to be 99.79%, suggesting that the method is simple, accurate, and reproducible. The proposed method proved to be sensitive, accurate, precise, economical, and less time-consuming, making it suitable for successful analysis in microsphere formulations. [ABSTRACT FROM AUTHOR]

The relevance of this study lies in the lack of reliable information on the key aspects of the molecular mechanisms of fatty hepatosis, a common mammalian disease, and its dangerous complications, such as fibrosis, cirrhosis, and hepatocellular carcinoma. These circumstances substantially complicate the prompt diagnosis of this hepatopathology and reduce the effectiveness of treatment. The metabolism of the heme derivative bilirubin is unique and inherent in the liver, which undergoes transformation due to hepatocyte enzyme systems to form neutral compounds, such as glucurono-conjugates and sulphoconjugates, which undergoes changes and requires analysis in fatty hepatosis in animals. The aim of the study was to determine the characteristic changes in the content of unconjugated bilirubin and its sulphoconjugate, glucuronide, monoglucuronide, monoglucoside, and diglucuronide in the blood, bile, and liver of rats under the modelling of fatty hepatosis and the administration of rehabilitation therapy. For this, the thin-layer chromatography method was used. It was found that under tetracycline-induced fatty hepatosis in laboratory rats, the concentration of unconjugated bilirubin in the blood decreased by 39.3%, bilirubin glucuronide by 44.4%, and the total fraction of bilirubin monoglucuronide and monoglycoside by 78.9%. Oral administration of the milk phospholipid complex in the composition of the BAS "FLP-MD" to sick animals ensured the maintenance of these indicators at the control level. The content of bilirubin glucuronide and bilirubin diglucuronide decreased in the liver tissue of the treated rats. When these animals were administered a phospholipid-containing bioadditives, the level of unconjugated bilirubin in the liver tissue increased by 22.2%, but the content of its conjugated forms decreased. The content of all the studied bilirubin fractions decreased in the bile of sick rats during self-rehabilitation. The components of the BAS "FLP-MD" in fatty hepatosis in rats did not affect the reduction of unconjugated bilirubin in bile but eliminated the inhibitory effect of the modelled pathology on the content of all conjugated forms of bilirubin. The findings can be used as biochemical markers to monitor the pigment metabolism in the development of fatty hepatosis in mammals. [ABSTRACT FROM AUTHOR]

This study investigated the insecticidal efficacy of essential oils (EOs) extracted from Ageratum conyzoides, Hyptis suaveolens, and Ocimum basilicum against female Anopheles mosquitoes, aiming to explore their potential as alternatives to synthetic insecticides amidst rising resistance issues. EOs were obtained through steam distillation from freshly harvested plant aerial parts, and their chemical compositions were analyzed using gas chromatography-mass spectrometry. The results revealed significant variations in chemical profiles among the oils, with precocene I dominating in A. Conyzoides, eucalyptol in H. suaveolens, and estragole in O. basilicum. Thin layer chromatography analyses revealed various components with rf values ranging from 0.25 – 0.93. Mosquito bioassay demonstrated varying knockdown effects across the oils, with H. suavolens achieving 77.5% knockdown within six minutes of the observation period. None of the oils or their combinations reached susceptibility status (98–100% mortality), indicating prevalent resistance among the mosquito population in the study area. The combination of A. conyzoides and H. suavolens essential oil gave the highest percentage mortality (70%) at the least time (9 minutes), this is a suggestion of synergistic activity. Despite resistance challenges, this study highlights the promise of botanical insecticides in sustainable mosquito control and underscores the ongoing need for innovation and adaptation in vector management strategies.

The article discusses the peculiarities of forensic examination of nalbuphine hydrochloride, a prescription medicine that can be purchased without a prescription in many pharmacies. At present, in Ukraine, nalbuphine is not subject to subject-quantitative accounting in healthcare facilities and by pharmaceutical manufacturers, but in some countries it is included in the list of narcotic drugs. It is emphasised that nalbuphine hydrochloride is a synthetic opioid analgesic, which is chemically similar to morphine and phenanthrene. In terms of pharmacological action, it belongs to the group of opioid receptor agonists-antagonists (pentazocine, buprenorphine, butorphanol). The article provides a brief overview of the effectiveness of nalbuphine for pain relief in various cases, as well as examples of abuse of opioids in combination preparations and facts of non-medical use of nalbuphine. The effectiveness of opioids as painkillers is undeniable, but today, given the risk/benefit ratio, the expediency of their long-term use is questionable. There are more and more recommendations to avoid the use of opioids. Nalbuphine in combination with other psychotropic substances, medicines and in large doses is life-threatening, so it is proposed to further study and control it more deeply at the state level by amending the Resolution of the Cabinet of Ministers of Ukraine “On Approval of the List of Narcotic Drugs, Psychotropic Substances and Precursors” of 6 May 2000 No. 770, in particular, to classify nalbuphine as a narcotic drug. The article analyses the peculiarities of sampling and sample preparation, the main stages of chemical research of this potentially dangerous opioid using thin-layer chromatography, infrared spectroscopy, gas chromatography with a mass-selective detector. It is emphasised that the experimental data presented in the article will help in choosing the most appropriate methods and conditions for the study of nalbuphine, and will allow an objective assessment of the results of expert research.

Lipidomic analysis in diverse biological settings has be-come a frequent tool to increase our understanding of the processes of life. Cellular lipids play important roles not only as being the main components of cellular membranes, but also in the regulation of cell homeostasis as lipid signaling molecules. Yeast has been harnessed for biomedical research based on its good conservation of genetics and fundamental cell organisation principles and molecular path-ways. Further application in so-called humanised yeast models have been developed which take advantage of yeast as providing the ba-sics of a living cell with full control over heterologous expression. Here we present evidence that high-performance thin-layer chroma-tography (HPTLC) represents an effective alternative to replace cost intensive mass spectrometry-based lipidomic analyses. We provide statistical comparison of identical samples by both methods, which support the use of HPTLC for quantitative analysis of the main yeast lipid classes.

Introduction: In Ayurveda, Shodhana is a method for purification or detoxification of some herbal drugs, minerals, and metals. Shodhana helps to remove or reduce the toxic properties of herbal drugs and enhances their therapeutic efficacy. Semecarpus anacardium Linn seeds (SA seeds) commonly known as Bhallataka is an important medicinal herb in Ayurveda. As the raw drug causes toxicity, it has to undergoes the Shodhana process to neutralize its toxic properties. Methods: In the present work, we studied transformation in chemical profiling of Shoodhit and Ashodhit Bhallataka (SA L.f) using high-performance thin-layer chromatography (HPTLC) profiling, thin-layer chromatography profiling, proton nuclear magnetic resonance (1H-NMR) fingerprinting, physicochemical analysis, and estimation of total phenolic and total flavonoids. The SA after Shodhana or purification process showed lesser oil contents as compared to raw drugs. Results: The Shodhana process led to the significant reduction of toxic fatty acid or oil content and reduction in the toxic phenolic components in the processed drug of SA L.f. The HPTLC profiling and 1H-NMR fingerprinting showed remarkable changes in the chemical profiling between the Shodhit and Ashodhit samples. The total phenolic and flavonoid content was found to be higher in unprocessed SA seeds than in the purified SA seeds. Conclusion: The study helps in the identification of the transformation of SA L.f. seeds concerning various chemical and physical attributes. The method used in the traditional medicine systems efficiently detoxify the raw drug of SA seeds. These approaches can be used in quality control analysis to ensure the effectiveness of Shodhana process.

Background: Many effective formulations are available in Ayurveda for various diseases. These formulations are lagging in standardization due to the absence of reference standards, whereas maintaining quality standards of given medicines is the need of an hour. Daruharidradi Ghanvati is one such combination of six herbal drugs containing Daruharidra, Meshshrungi, Vijaysar, Mamajjak, Jambubija, and Methikabij. Each drug is described in various Ayurvedic antidiabetic formulations. Aim: The present study was aimed at setting a standard pharmacognostical and pharmaceutical profile of Daruharidradi Ghanavati. Materials and Methods: The study included the preparation of Daruharidradi Ghanavati using raw drugs. Later, Daruharidradi Ghanavati was subjected to pharmacognostical, physiochemical, and thin-layer chromatography analysis as per standard protocols. Results: The final observations were recorded. Pharmacognostical findings matched with that of individual raw drugs with no major change in the microscopic structure of the raw drugs during the preparation of Ghanavati. Conclusion: The quality of Daruharidradi Ghanavati tablet can be tested by pharmacognostical, physiochemical screening for the observations of the present study. [ABSTRACT FROM AUTHOR]

IntroductionKefir beverage has beneficial microorganisms that have health-giving properties; therefore, they have a good potential to be probiotic. This study evaluated the probiotic potential, technological, and safety characteristics of Enterococcus faecalis, Lactococcus lactis, and Pichia fermentans isolated from traditional kefir beverages.MethodFirst, isolates were evaluated in terms of resistance to acid, alkali, bile salts, trypsin, and pepsin of the gastrointestinal tract. The auto-aggregation and co-aggregation ability of isolates were measured using spectrophotometry. Antimicrobial activities were assayed against important food-borne pathogens using the agar well diffusion method. Moreover, gamma-aminobutyric acid (GABA) production was investigated by thin-layer chromatography (TLC).ResultAmong the isolates, P. fermentans had an 85% total survival rate, but its amount reached below 6 log CFU/ml which is considered non-resistant, and it showed the highest auto-aggregation (74.67%). Moreover, only L. lactis showed antimicrobial activity and had the highest co-aggregation with E. coli PTCC 1338 (54.33%) and L. monocytogenes ATCC 7644 (78%). Finally, an evaluation of the technological and safety characteristics of the strains showed that the strains produced GABA and were safe.DiscussionAlthough the isolates were not resistant to the gastrointestinal tract, their supernatant contained valuable natural compounds, including antioxidants, GABA, and antimicrobials, which can be used to produce functional foods and medicines. In addition, other approaches, such as increasing the initial number of strains, using foods as carriers of isolates, and encapsulating the isolates, can effectively increase the survivability of isolates in the gastrointestinal tract.

To establish a reliable method for measuring the radiochemical purity of technetium[99mTc] methylenediphosphonate injection, analysis conditions such as stationary phase and developing solvent were studied. The analytical method was methodologically verified in accordance with the Chinese Pharmacopoeia and other relevant provisions. The results showed that the analytical method of single-system thin-layer chromatography, in which polyamide-6 thin-layer plate was used as the stationary phase, V (1 mol/L sodium acetate solution): V (0.1 mol/L hydrochloric acid solution) =1∶1 mixed solution was used as the developing solvent, could improve the tailing problem of the main peak of technetium[99mTc] methylenediphosphonate injection. The resolution of radioactive main peak and potential impurities including pertechnetate[99mTc] and colloidal technetium[99mTc] was greater than 1.0. The method had a good linear relationship in the radioactivity range of 2.7×10-2 μCi to 42.4 μCi, and the correlation coefficient r was 0.997, with good specificity, good durability and intermediate precision. Compared with the analytical method of the pharmacopoeias, the relative error of radiochemical purity met the requirement of less than 2.0%. The established thin-layer chromatography is suitable for determination of the radiochemical purity of technetium[99mTc] methylenediphosphonate injection.

Scientific relevance. Cardiac glycosides have been used in medicine for over two centuries. Current studies suggest that biologically active substances from this group can be used to treat not only heart conditions but also viral infections, cancers, and other diseases. Therefore, quality control methods for cardiac glycosides are becoming increasingly relevant.Aim. Based on a review of Russian and international quality standards, as well as up-to-date scientific data, this study aimed to identify promising methods for the identification and quantification of cardiac glycosides in herbal drugs and herbal medicinal products, as well as to evaluate the possibility of substituting physicochemical methods for biological methods.Discussion. The methods that are currently used to standardise cardiac glycosides are either not selective or require laboratory animals (biological test systems). According to a study of pharmacopoeial methods for the identification of cardiac glycosides in herbal drugs and herbal medicinal products, chemical identification tests and thin-layer chromatography continue to be relevant. Quantitative testing of herbal drugs and extracts uses biological and non-selective (spectrophotometry) methods, whereas chromatography is described only in general and individual monographs for herbal drug preparations containing individual cardiac glycosides and medicinal products containing these preparations. Upon analysing quality standards and scientific publications, the authors identified potentially promising methods for the quantification of cardiac glycosides in herbal drugs, herbal drug preparations, and herbal medicinal products, namely chromatographic methods.Conclusions. Reverse-phase high-performance liquid cjromatography (HPLC) with spectrophotometric detection is the most suitable method for pharmacopoeial analysis. The development of an HPLC-based analytical procedure to determine the cardiac glycoside content will provide an opportunity to advance from biological or non-selective methods to more ethical and selective up-to-date techniques.

This study presents the establishment of surface-enhanced Raman spectroscopy (SERS) combined with the thin-layer chromatography (TLC) method for detecting four types of quinolone antibiotics in aquatic products. The experimental procedure involved ultrasonic extraction using methanol as the extraction solvent to isolate the target ingredients. The quinolone antibiotics were separated using chromatography conditions with Merck silica gel G TLC plate as a stationary phase and a chloroform – methanol – ethyl acetate – ammonia solution (5 : 12 : 1 : 4) as a mobile phase. Silver sol was used as the matrix to obtain SERS peaks for quinolone. The quinolone antibiotics were identified using SERS in aquatic products. Subsequently, normal SERS spectra of quinolone antibiotic powder, quinolone antibiotic preparations, SERS characteristic peaks, and quinolone antibiotic in fish and shrimp were analyzed. The TLC chromatography system allowed to achieve the separation of four quinolone preparations. The system suitability test showed that the humidity range was 18–88%, and the durability range was between 20–50°C. Moreover, the established TLC-SERS method successfully identified quinolones in aquatic products. Therefore, the SERS method could achieve the effective qualitative determination of quinolones, offering high sensitivity, simple operation, and high timeliness, and is suitable for the rapid detection of quinolones. [ABSTRACT FROM AUTHOR]

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The rising discharge of anthropogenic chemicals into aquatic environments poses a significant threat, necessitating effective monitoring strategies. This study introduces an innovative approach to effect-directed analysis (EDA) by coupling liquid chromatography (LC) with high-performance thin-layer chromatography (HPTLC), utilizing a modified MALDI spotter. The objective is to optimize fractionation parameters for sample application and assess the method's viability in identifying acetylcholinesterase (AChE) inhibitors, specifically malathion, parathion, and chlorfenvinphos.The optimization process involves controlling sample volume, spot shape, and spot distances on HPTLC plates. Successful application is demonstrated by a miniaturized LC system coupled to the HPTLC plate via spotter, allowing effective separation and identification of AChE inhibitors. The study further explores the method's application to water samples from a river with predominantly agricultural drainage area.Spiked samples reveal distinct active spots, identified through extraction and subsequent high-resolution mass spectrometry (HRMS) measurements. However, results indicate the absence of AChE inhibitors in non-spiked water samples, affirming the efficacy of the EU ban on most organophosphate pesticides. The usefulness of HPTLC in separation of coeluting substances from HPLC is emphasized, demonstrating its suitability for the effect-directed analysis of complex samples.This work shows the integration of HPTLC with liquid chromatography for EDA, offering a powerful tool for identifying and monitoring AChE inhibitors in water samples. The approach addresses limitations in current monitoring strategies and provides insights into the presence and impact of chemicals in aquatic ecosystems. The study contributes to ongoing efforts to enhance water quality monitoring, aligning with the principles of the EU Water Framework Directive.

Before formulating radiopharmaceuticals for injection, it is necessary to remove various impurities via purification. Conventional synthesis methods involve relatively large quantities of reagents, requiring high-resolution and high-capacity chromatographic methods (e.g., semi-preparative radio-HPLC) to ensure adequate purity of the radiopharmaceutical. Due to the use of organic solvents during purification, additional processing is needed to reformulate the radiopharmaceutical into an injectable buffer. Recent developments in microscale radiosynthesis have made it possible to synthesize radiopharmaceuticals with vastly reduced reagent masses, minimizing impurities. This enables purification with lower-capacity methods, such as analytical HPLC, with a reduction of purification time and volume (that shortens downstream re-formulation). Still, the need for a bulky and expensive HPLC system undermines many of the advantages of microfluidics. This study demonstrates the feasibility of using radio-TLC for the purification of radiopharmaceuticals. This technique combines high-performance (high-resolution, high-speed separation) with the advantages of a compact and low-cost setup. A further advantage is that no downstream re-formulation step is needed. Production and purification of clinical scale batches of [18F]PBR-06 and [18F]Fallypride are demonstrated with high yield, purity, and specific activity. Automating this radio-TLC method could provide an attractive solution for the purification step in microscale radiochemistry systems.

Scientific relevance. The State Pharmacopoeia of the Russian Federation, edition XIV, requires the identification of aniseed and fennel fruits by the same thin-layer chromatography (TLC) procedure using Sudan III and menthol. These markers are neither specific nor related to the therapeutic effects of these herbal drug products. The visual interpretation of chromatograms is complicated because of the low intensity of adsorption zones. Moreover, the corresponding compendial monographs do not characterise the adsorption zones specific to each of the plants. The most abundant component in aniseed and fennel essential oils, trans-anethole, would make a better reference standard from a methodological point of view.Aim. This study aimed to optimise the TLC procedure for essential oil determination in herbal drugs and herbal medicinal products of aniseed and fennel fruits and subsequently recommend it for inclusion in the relevant compendial monographs.Materials and methods. The study examined samples from several batches of herbal drugs, including aniseed and common fennel fruits, sourced from Russian manufacturers. The reference standards comprised commercial reagents, including trans-anethole, anise oil, and linalool, as well as fresh essential oils that had been steam distilled from the test samples in a Clevenger apparatus. The study was conducted by TLC. Sample preparation involved using Merck aluminium TLC plates, an IKA KS-501 digital orbital shaker, and a CAMAG Linomat 5 semi-automatic sample application system. The authors heated the plates in either a Binder ED53 drying oven or a CAMAG TLC plate heater. For visualisation, the authors selected a CAMAG TLC VISUALIZER 2 UV imaging and documentation system.Results. The informational and experimental research showed the feasibility of using trans-anethole as an identification reference standard for common anise and fennel fruits. The authors selected the optimal solvent for extracting active substances from the test products (hexane) and a detection reagent for improving visual evaluation of the chromatograms (1% sulfovanillin). The authors established characteristic adsorption zones for differentiating between aniseed and fennel fruits.Conclusions. The optimised procedure identifies the main components in the essential oils of aniseed and fennel fruits with a specific reference standard. This procedure may be recommended for inclusion in draft monographs for the corresponding herbal medicinal products.

Scientific relevance. When harmonising conditions for qualitative and quantitative analysis of bioactive compounds in herbal medicines, the Institute of Pharmacopoeia and Medicinal Product Standardisation has discovered that horse-chestnut products lack national quality standards.Aim. This study aimed to review international quality standards in order to identify the most promising testing methods for the main groups of bioactive compounds in horse-chestnut seeds and draw upon these methods when drafting national pharmacopoeial monographs for horse-chestnut seed products.Discussion. The authors compared the requirements established by leading world pharmacopoeias for the identification and assay of herbal drugs and herbal drug preparations derived from horse-chestnut seeds.Conclusions. The study results show that, predominantly, identification tests are based on thin-layer chromatography, assays rely on spectrophotometry, and aescin is used as a pharmacopoeial reference standard. The authors drafted a pharmacopoeial monograph for the medicinal product Horse-Chestnut Seed Dry Extract + Thiamine Hydrochloride, Oral Solution, which requires using thin-layer chromatography and spectrophotometry for qualitative and quantitative testing.

To date, the aetiology and molecular mechanisms of the development of fatty hepatosis, which is quite common in mammals, have not yet been sufficiently explained. This pathology requires detailed study not only because of functional disorders of the liver and biliary system, but also because of the high probability of dangerous complications – fibrosis, cirrhosis, or hepatocellular carcinoma. The purpose of this study was to determine marker changes in the phospholipid composition of bile and blood in rats with experimental fatty hepatosis and with the use of corrective therapy. Hepatopathology was modelled in Wistar laboratory rats by intragastric administration of a 4% solution of tetracycline hydrochloride at the rate of 0.25 g/kg body weight for seven days. Using the method of thin-layer chromatography, the phospholipid components of animal bile and blood were studied. It was found that during experimental fatty hepatosis in rats, there was a decrease in the total phospholipid content in bile, mainly due to a decrease in the level of phosphatidylcholine (by 22.4-27.0%), the total fraction of inositol phosphatide and phosphatidylinositol (by 20.0-27.3%), and phosphatidylethanolamine (by 17.5-25.2%). Conversely, the introduction of milk phospholipids in the form of a dietary supplement “FLP-MD” in sick animals contributed to an increase in bile levels of phosphatidylserine by 67.1-99.8%, inositol phosphatide and phosphatidylinositol by 48.6-57.6%, phosphatidylcholine by 38.8-60.2%, phosphatidylethanolamine by 45.6-57.4%, and sphingomyelin by 30.4-46.3%. In the blood of such rats, a significant decrease in the content of phosphatidylcholine, phosphatidylserine, and sphingomyelin was found, which was not observed after administration of the “FLP-MD” dietary supplement to sick animals. In the case of using the supplement in healthy animals, only a 29.3% increase in the blood content of inositol phosphatide and phosphatidylinositol was noted. The determination of the most sensitive indicators in the phospholipid spectrum of blood and bile reveals the features of changes in molecular processes for the development of fatty hepatosis in animals, and also contributes to preclinical tests of the corrective effectiveness of newly created drugs according to established markers

Background and Aim: Antibiotic residues in livestock farming have been identified as a potential cause of antimicrobial resistance in humans and animals. This study aimed to determine whether antibiotic residues were present in the chicken meat, eggs, feces, and feed collected from all four districts in the Mymensingh division of Bangladesh. Materials and Methods: To detect antibiotic residues in the collected samples, qualitative thin-layer chromatography (TLC) and quantitative high-performance liquid chromatography (HPLC) were used. A total of 230 samples were analyzed for commonly used 11 antibiotics residue. Out of these, 40 meats and 40 feces samples were collected from broilers and layers, 30 egg samples from duck and layer, and 120 feed samples from both broilers and layers from the study area. Thin-layer chromatography was used for screening the presence of antibiotic residues; TLC-positive samples were then subjected to further HPLC analysis to determine the residue concentrations. Results: Thin-layer chromatography analysis revealed that 23.5% of the tested samples contained residues from six different antibiotic classes (tetracyclines, quinolones, beta-lactams, sulfonamides, aminoglycosides, and macrolides). Thin-layer chromatography analysis showed that 35% and 25% of the meat samples were positive for residues from the broiler and layer, respectively. About 15% and 30% of layer and duck egg samples had positive residues, respectively. Out of 120 feed samples analyzed, about 15.8% had various antibiotic residues. In addition, feces samples from broilers and layers had 50% and 35% antibiotic residues, respectively. A total of 2.5% meat and 3.3% egg samples had antibiotic residues above the maximum residue limit (MRL). Based on the findings of this study, the highest percentage of oxytetracycline, followed by doxycycline and ciprofloxacin, were detected in feed samples, and oxytetracycline was detected in meat and egg samples. Conclusion: This study clearly showed the misuse of antibiotics in the poultry sector in Bangladesh. Although antibiotic residues below the MRL level are suitable for human consumption, they may result in antimicrobial drug resistance to pathogens.

Oaks (Quercus L., Fagaceae) are a widespread tree species worldwide, and in Hungary they account for nearly 30 % of the forests. Their wood is valuable, but their bark is considered as a by-product. Oak bark, available in large quantities but with no dedicated use, contains a significant amount of valuable extractives. Its (+)-catechin content is around 1 %. (+)-Catechin is mostly used for food industry, medicine and many other industrial purposes, representing a significant financial value. The aim of the present research was to compare the (+)-catechin concentrations in the bark of the most important oak species found in Hungary and to optimize sample pretreatment (conservation) and extraction methods in order to achieve fast and efficient extraction. From these species the highest concentrations were measured in Q. robur and Q. robur ssp. slavonica (8–12 mg (+)-catechin/g dry bark). The combination of microwave sample pretreatment and microwave assisted extraction proved to be the most time- and cost-effective method. The utilization of the extracted bark powder for energetic purposes requires further investigations.

In recent years, the interest in natural remedies has increased, so it is important to analyze the plants widely distributed in nature but whose composition is little known. The main objective of the present work is to obtain information based on the profiles of secondary metabolites and antioxidant activity in Lamium album, a very widespread but little studied plant, with the aim of revealing the differences compared to Urtica dioica. First, the optimization of enzymatic extraction assisted by ultrasound was carried out by the Box–Behnken method. The optimized parameters were: concentration of the enzyme—3.3% cellulase, temperature—55 °C, and the extraction time—40.00 min. The efficiency was estimated based on the content of iridoids, the main class of secondary metabolites from Lamium album. Second, the secondary metabolites profiles of the nettle extracts were obtained by thin-layer chromatography using both normal and reverse phases and by RP-UHPLC. The antioxidant activity was evaluated using DPPH and ABTS+ radicals. The obtained results revealed significant differences between the two nettle species, both in terms of the phytochemical compounds, as well as the antioxidant activity, confirming the fact that Lamium album has a high potential to be used in phytomedicine. [ABSTRACT FROM AUTHOR]

Enantiomeric separation of (RS)-Metolazone has been achieved by thin-layer chromatography (TLC) using both direct modes. cyclodextrin, containing hydroxylic group, was utilized as the chiral selector. For enantioseparation, cyclodextrin was used as a chiral additive in the stationary phase in a noncovalent mode and there was no chiral additive in the mobile phase; the native enantiomers were isolated and characterized. Cyclodextrin was also added to mobile phase and there was no chiral selector in the stationary phase. The spots were then isolated and characterized. The effect of the composition of the mobile phase, pH, and temperature was studied for the optimization of successful separation conditions. The spots were located in an iodine chamber. [ABSTRACT FROM AUTHOR]

Background and Aim: Antibiotic residues in livestock farming have been identified as a potential cause of antimicrobial resistance in humans and animals. This study aimed to determine whether antibiotic residues were present in the chicken meat, eggs, feces, and feed collected from all four districts in the Mymensingh division of Bangladesh. Materials and Methods: To detect antibiotic residues in the collected samples, qualitative thin-layer chromatography (TLC) and quantitative high-performance liquid chromatography (HPLC) were used. A total of 230 samples were analyzed for antibiotic residues of commonly used 11 antibiotics. Out of these, 40 meat and 40 feces samples were collected from broilers and layers, 30 egg samples from ducks and layers, and 120 feed samples from broilers and layers from the study area. Thin-layer chromatography was used to screen the presence of antibiotic residues; TLC-positive samples were then subjected to further HPLC analysis to determine the residue concentrations. Results: Thin-layer chromatography analysis revealed that 23.5% of the tested samples contained residues from six different antibiotic classes (tetracyclines, quinolones, beta-lactams, sulfonamides, aminoglycosides, and macrolides). Thin- layer chromatography analysis showed that 35% and 25% of the meat samples were positive for residues from the broiler and layer, respectively. About 15% and 30% of layer and duck egg samples had positive residues, respectively. Out of 120 feed samples analyzed, about 15.8% had various antibiotic residues. In addition, feces samples from broilers and layers had 50% and 35% antibiotic residues, respectively. A total of 2.5% meat and 3.3% egg samples had antibiotic residues above the maximum residue limit (MRL). Based on the findings of this study, the highest percentage of oxytetracycline, followed by doxycycline and ciprofloxacin, were detected in feed samples, and oxytetracycline was detected in meat and egg samples. Conclusion: This study clearly showed the misuse of antibiotics in the poultry sector in Bangladesh. Although antibiotic residues below the MRL level are suitable for human consumption, they may result in antimicrobial drug resistance to pathogens. [ABSTRACT FROM AUTHOR]

To date, the aetiology and molecular mechanisms of the development of fatty hepatosis, which is quite common in mammals, have not yet been sufficiently explained. This pathology requires detailed study not only because of functional disorders of the liver and biliary system, but also because of the high probability of dangerous complications - fibrosis, cirrhosis, or hepatocellular carcinoma. The purpose of this study was to determine marker changes in the phospholipid composition of bile and blood in rats with experimental fatty hepatosis and with the use of corrective therapy. Hepatopathology was modelled in Wistar laboratory rats by intragastric administration of a 4% solution of tetracycline hydrochloride at the rate of 0.25 g/kg body weight for seven days. Using the method of thin-layer chromatography, the phospholipid components of animal bile and blood were studied. It was found that during experimental fatty hepatosis in rats, there was a decrease in the total phospholipid content in bile, mainly due to a decrease in the level of phosphatidylcholine (by 22.4-27.0%), the total fraction of inositol phosphatide and phosphatidylinositol (by 20.0-27.3%), and phosphatidylethanolamine (by 17.5-25.2%). Conversely, the introduction of milk phospholipids in the form of a dietary supplement "FLP-MD" in sick animals contributed to an increase in bile levels of phosphatidylserine by 67.1-99.8%, inositol phosphatide and phosphatidylinositol by 48.6-57.6%, phosphatidylcholine by 38.8-60.2%, phosphatidylethanolamine by 45.6-57.4%, and sphingomyelin by 30.4-46.3%. In the blood of such rats, a significant decrease in the content of phosphatidylcholine, phosphatidylserine, and sphingomyelin was found, which was not observed after administration of the "FLP-MD" dietary supplement to sick animals. In the case of using the supplement in healthy animals, only a 29.3% increase in the blood content of inositol phosphatide and phosphatidylinositol was noted. The determination of the most sensitive indicators in the phospholipid spectrum of blood and bile reveals the features of changes in molecular processes for the development of fatty hepatosis in animals, and also contributes to preclinical tests of the corrective effectiveness of newly created drugs according to established markers. [ABSTRACT FROM AUTHOR]

In this work, a screen-printing method was developed to create porous particle-based materials as layers with specifically designed shape to produce microfluidics systems. Among several tested binding agents, xanthan gum was found to be an excellent choice for a printing mixture thickener as well as a durable binder for the resulting material. In addition to demonstrating control over the shape of the printed microfluidics chips, control over material thickness, wetting characteristics and general method accuracy were also investigated. The applicability of the introduced method was further demonstrated with a development of an exemplary microfluidics chip for quantitative detection of Fe (III), Ni (II), Cu (II), Cd (II), and Pb (II) from a mixed sample at millimolar levels. The novel approaches demonstrated in this article offer new perspective into creating multiplexed on-site chemical analysis tests. [ABSTRACT FROM AUTHOR]

Sorghum is a unique natural food source of 3-deoxyanthocyanidins (3-DA) polyphenols. This work evaluated the effect of acidification on sorghum extracts post pressurized liquid extraction (PLE) and its ability to increase the identification and quantification of 3-DA. The sorghum genotypes included Sumac and PI570366 (bran only) and SC991 (leaf and leaf sheath tissue). The acidification of the PLE extracts was carried out with methanol–HCl solutions at various concentrations (0, 0.5, 1, 2, and 4%, v/v). Changes in color were determined using L*a*b*. The overall phenolic composition was estimated with the total phenolic content and the DPPH free radical scavenging assays. Quantitative and qualitative chromatographic methods determined the phenolic profile. Color analysis showed that the redness and color saturation increased after acidification. No statistical difference was found in the total phenolic content of the acidified extracts, except for SC991, which was increased. There were no differences in the antioxidant capacity following acidification in all samples. For chromatographic analysis, luteolinidin was predominant in the extracts and the 3-DA content increased after acid treatment. However, some flavonoid and phenolic acid concentrations decreased following acid treatment, including taxifolin, quercetin, and chlorogenic acid. Interestingly, 0.5% v/v HCl acidification was sufficient to increase the color, allow the detection of 5-methoxyluteolinidin, and to increase luteolinidin and 7-methoxyapigenidin by at least twofold. [ABSTRACT FROM AUTHOR]

Introduction: Ayurvedic formulations have a long history of use and have increased amazingly to follow the global interest in natural products. However, for many classical ayurvedic formulations, no quality parameters of standardization are available. Vyoshadi Churna, a classical Ayurvedic polyherbal formulation that cures indigestion and diarrhea, is being used since ancient times. However, there is no standard method available for its formulation and evaluation. Hence, the current study aims to develop the quality standards for Vyoshadi churna. Methods: The standard procedures for preparation and standardization of Churna described in Ayurvedic Pharmacopoeia of India (API) were used in the present study. Standardization includes pharmacognostical, chromatographic, and physico-chemical studies. The other essential studies were also done in accordance to API and World Health Organization guidelines to find the limits of microbial load, aflatoxin, heavy metals, and pesticide residue using modern analytical techniques such as gas chromatography-mass spectroscopy (MS) and inductively coupled plasma-MS/MS. Results: Vyoshadi Churna is of brown color and bitter in taste. Pharmacognostical studies have shown the characteristic features of almost each contributory crude drug. A unique chemical profile was also developed using toluene: ethyl acetate: methanol: formic acid (4:4:1:0.5) as the mobile phase to resolve maximum components. Furthermore, the standardized limits for the physicochemical studies, microbial load, heavy metals, aflatoxins, and pesticide residues were established and found in limits. Conclusion: This is for the first time when the standard operating procedure of Vyoshadi Churna was developed. Hence, the current technical work would be practically helpful for the future preparation and standardization study of Vyoshadi Churna.

Sida rhombifolia belongs to the Malvaceae family and is often used to treat gout in Indonesia. S. rhombifolia has many efficacies and contains many different chemical components. The abundance and variation of chemical content and chemical compounds in this medicinal plant are challenging factors in ensuring medicinal plants' safety and quality control. Thin-layer chromatography (TLC) fingerprint analysis derived from S. rhombifolia extract can also be used for the medicinal plant's quality control. This research aimed to develop the optimum condition for the chemical fingerprint analysis of S. rhombifolia using a TLC fingerprint analysis. A total of 11 bands were produced with optimum separation using silica gel 60 F254 TLC plate, a mixed mobile phase condition with chloroform, ethyl acetate, and methanol (6.5:2:1.5). This fingerprint analysis performed an excellent separation in the TLC plate at 366 nm with sulfuric acid as reagent derivatization. In general, the results of the analysis validation, including stability, specificity, precision, and robustness of TLC fingerprints, met the acceptance criteria. The TLC fingerprint of S. rhombifolia can be distinguished from 2 related plants with similar leaf shapes, Turnera ulmifolia L. and Hibiscus rosa-sinensis. The developed method was validated, so it could be used to control S. rhombifolia quality.

IntroductionThe need for new tools to treat infections is constantly growing due to the possibilities of emerging diseases related to environmental changes, climatic catastrophes, microorganism resistance, and human and animal aging, leading to an evident unbalance in the planet’s health. Brazil contains the most significant portion of world biodiversity, a potential source of new antimicrobial natural products. Nonetheless, its environment, particularly its forests, and rainforests, is under threat, meaning that rapidly conducted, comprehensive research into the potential of antimicrobial activity to address this threat is urgently needed.MethodsIn this study, plants from the Amazon rainforest and the Atlantic forests were collected and tested against several pathogenic microbes relevant to humans, animals, and the environment, and subjected to large-scale susceptibility assays, bioautography, and Artemia salina toxicity assays. From the plants, 2,280 organic and aqueous extracts were obtained from different organs, namely leaves, barks, flowers, fruits, and seeds, and subjected to a large-scale susceptibility screening assay against Staphylococcus aureus, Staphylococcus epidermidis, Enterococcus faecalis, Streptococcus mutans, Streptococcus sanguinis, Escherichia coli, Pseudomonas aeruginosa, Candida albicans, Malassezia pachydermatis, Malassezia furfur, and Listeria monocytogenes. Results and discussionThe selected extracts were subjected to antimicrobial susceptibility tests to determine their inhibition zone diameters and minimum bactericidal concentrations, to bioautography, and to an Artemia salina toxicity assay, which resulted in 154 active extracts. Moreover, 111 out of 154 extracts were ranked based on scores established by the p-values and the mean rank differences in each set of test results. The final ranking identified which extracts should be studied in further phytochemical research using thin-layer chromatography techniques as a priority. The extracts obtained from plants belonging to Combretaceae, Connaraceae, Convolvulaceae, Fabaceae, Malpighiaceae, Moraceae, Piperaceae, Polygonaceae, and Salicaceae were selected as the most promising ones and used to support the identification of plant-based antimicrobial active compounds from the immense biodiversity of Brazilian forests.

The advantages of hyperspectral imaging in videodensitometry are presented and discussed with the example of extracts from 70 Polish grasses. An inexpensive microscope camera was modified to cover the infrared spectrum range, and then 11 combinations of illumination (254 nm, 366 nm, white light), together with various filters (no filter, IRCut, UV, cobalt glass, IR pass), were used to register RGB HDR images of the same plate. It was revealed that the resulting 33 channels of information could be compressed into 5–6 principal components and then visualized separately as grayscale images. We also propose a new approach called principal component artificial coloring of images (PCACI). It allows easy classification of chromatographic spots by presenting three PC components as RGB channels, providing vivid spots with artificial colors and visualizing six principal components on two color images. The infrared region brings additional information to the registered data, orthogonal to the other channels and not redundant with photos in the visible region. This is the first published attempt to use a hyperspectral camera in TLC and it can be clearly concluded that such an approach deserves routine use and further attention. [ABSTRACT FROM AUTHOR]

IntroductionRadio thin layer chromatography (radio-TLC) is commonly used to analyze purity of radiopharmaceuticals or to determine the reaction conversion when optimizing radiosynthesis processes. In applications where there are few radioactive species, radio-TLC is preferred over radio-high-performance liquid chromatography due to its simplicity and relatively quick analysis time. However, with current radio-TLC methods, it remains cumbersome to analyze a large number of samples during reaction optimization. In a couple of studies, Cerenkov luminescence imaging (CLI) has been used for reading radio-TLC plates spotted with a variety of isotopes. We show that this approach can be extended to develop a high-throughput approach for radio-TLC analysis of many samples.MethodsThe high-throughput radio-TLC analysis was carried out by performing parallel development of multiple radioactive samples spotted on a single TLC plate, followed by simultaneous readout of the separated samples using Cerenkov imaging. Using custom-written MATLAB software, images were processed and regions of interest (ROIs) were drawn to enclose the radioactive regions/spots. For each sample, the proportion of integrated signal in each ROI was computed. Various crude samples of [18F]fallypride, [18F]FET and [177Lu]Lu-PSMA-617 were prepared for demonstration of this new method.ResultsBenefiting from a parallel developing process and high resolution of CLI-based readout, total analysis time for eight [18F]fallypride samples was 7.5 min (2.5 min for parallel developing, 5 min for parallel readout), which was significantly shorter than the 48 min needed using conventional approaches (24 min for sequential developing, 24 min for sequential readout on a radio-TLC scanner). The greater separation resolution of CLI enabled the discovery of a low-abundance side product from a crude [18F]FET sample that was not discernable using the radio-TLC scanner. Using the CLI-based readout method, we also observed that high labeling efficiency (99%) of [177Lu]Lu-PSMA-617 can be achieved in just 10 min, rather than the typical 30 min timeframe used.ConclusionsCerenkov imaging in combination with parallel developing of multiple samples on a single TLC plate proved to be a practical method for rapid, high-throughput radio-TLC analysis.

The principle of consistent, end-to-end standardisation builds upon the fact that qualitative and quantitative analysis of bioactive compounds in herbal drugs, herbal drug preparations, and corresponding herbal medicinal products involves the same methods of analysis. The possibility of such standardisation should be considered at the stage of specification development. The aim of the study was to explore approaches to analysing of Eucalyptus viminalis leaves and chlorophyllipt medicinal products, observing the principle of end-to-end standardisation. Materials and methods: the authors analysed Eucalyptus viminalis as a herbal drug (leaves), a herbal drug preparation (chlorophyllipt soft extract), and herbal medicinal products (Chlorophyllipt®, 1% alcohol solution for oral, cutaneous, rectal, and vaginal use; 2% oil solution for cutaneous and vaginal use; and 25 mg orodispersible tablets) by direct UV spectrophotometry, thin-layer chromatography (TLC), and qualitative reactions. Results: the study demonstrated the possibility of using a herbal reference standard of eucalimin to analyse the Eucalyptus viminalis herbal drug and chlorophyllipt preparations and medicinal products in different dosage forms by TLC. Conclusions: procedures for determination of total phenolic aldehydes, if included into specifications and production technology of chlorophyllipt preparations and products, ensure compliance with the principle of end-to-end standardisation and allow for quantitative determination and control of active ingredients throughout the product lifecycle: from the acceptance of raw materials, through in-process controls, to testing and monitoring of finished products’ quality.

Favipiravir (FAV) has become a promising antiviral agent for the treatment of COVID-19. Herein, a green, fast, high-sample-throughput, non-instrumental, and affordable analytical method is proposed based on surfactant-assisted dispersive liquid–liquid microextraction (SA-DLLME) combined with thin-layer chromatography–digital image colourimetry (TLC-DIC) for determining favipiravir in biological and pharmaceutical samples. Triton X-100 and dichloromethane (DCM) were used as the disperser and extraction solvents, respectively. The extract obtained after DLLME procedure was spotted on a TLC plate and allowed to develop with a mobile phase of chloroform:methanol (8:2, v/v). The developed plate was photographed using a smartphone under UV irradiation at 254 nm. The quantification of FAV was performed by analysing the digital images' spots with open-source ImageJ software. Multivariate optimisation using Plackett–Burman design (PBD) and central composite design (CCD) was performed for the screening and optimisation of significant factors. Under the optimised conditions, the method was found to be linear, ranging from 5 to 100 µg/spot, with a correlation coefficient (R2) ranging from 0.991 to 0.994. The limit of detection (LOD) and limit of quantification (LOQ) were in the ranges of 1.2–1.5 µg/spot and 3.96–4.29 µg/spot, respectively. The developed approach was successfully applied for the determination of FAV in biological (i.e., human urine and plasma) and pharmaceutical samples. The results obtained using the proposed methodology were compared to those obtained using HPLC-UV analysis and found to be in close agreement with one another. Additionally, the green character of the developed method with previously reported protocols was evaluated using the ComplexGAPI, AGREE, and Eco-Scale greenness assessment tools. The proposed method is green in nature and does not require any sophisticated high-end analytical instruments, and it can therefore be routinely applied for the analysis of FAV in various resource-limited laboratories during the COVID-19 pandemic. [ABSTRACT FROM AUTHOR]

The aim of this work is to develop a device based on thin-layer chromatography coupled with surface-enhanced Raman spectroscopy (TLC-SERS) to analyze sulfamethoxazole (SMX) and trimethoprim (TMP) in commercial milk samples using chemometric tools. Samples were eluted in TLC plates, and a central composite design (CCD) of two factors was performed to optimize the gold nanoparticle dispersion on TLC plates for SERS, aiming at the detection of both drugs at concentrations close to their maximum residual limits (MRLs). Following the optimization, hyperspectral images from the SERS were captured of the TLC plates. Multivariate curve resolution (MCR-ALS) and independent component analysis (ICA) chemometric techniques were used to extract the signals of the analytes. All the samples presented recovery values of 81–128% for TMP. The quantification of SMX was not possible due to SERS suppression by an interferent. However, it was possible to detect SMX at a concentration of two times the MRL (8.0 × 10−7 mol·L−1). The results demonstrate that the TLC-SERS device is a potential tool for the quantification of TMP and the detection of SMX in milk. [ABSTRACT FROM AUTHOR]

To date, the world has accumulated a large amount of long-lived radioactive materials that need to be disposed of or reprocessed. Such materials include nuclear legacy objects containing 226Ra, which is an important material for obtaining a wide range of isotopes for nuclear medicine via irradiation in reactors, cyclotrons, and electron accelerators. For the selective recovery of 226Ra from waste materials, crown-ether (CE) 18-crown-6 (18C6) or its derivatives can be used, which, however, have not been widely studied for these purposes. In our work, the key property of 18C6 and its derivatives, the phase distribution, was studied using tritium labeling. The possibility of introducing a tritium label into CEs molecules using thermal activation of tritium has been demonstrated; a high specific activity of the obtained compounds was achieved (from 18 to 108 TBq/mol). Methods for chromatographic purification of the studied CEs were developed. The distribution of 18C6 and its derivatives between various organic solvents and water was studied in detail for the first time. Subsequently, the obtained data will allow us to choose conditions for the selective recovery of 226Ra from aged sources. [ABSTRACT FROM AUTHOR]

Molecular spectroscopy and photochemistry constitute an integral field in modern chemistry. However, undergraduate level classes provide limited opportunities for hands-on experimentation of photochemistry and photophysics. For this reason, a simple laboratory experiment was designed that may be easily implemented into undergraduate teaching laboratories with the aim of introducing undergraduate students to UV/visible spectroscopy and photochemistry/photophysics and its possible applications. Samples of three unknown sunscreen formulations are given to students and they are asked to use a set of techniques to identify their molecular composition and to test their efficacy using basic laboratory equipment available to them. In particular, the students are asked to complete the following tasks: (i) sample preparation using solvent extraction to extract active ingredients from the sunscreen lotion, (ii) identify the extracted molecular sunscreen constituents by Thin Layer Chromatography (TLC) and UV/visible spectroscopy, and finally (iii) study their photostability by means of steady state irradiation coupled with UV/visible spectroscopy. The students were provided with the following tools for data collection: silica-backed TLC plates, a short-wave lamp (254 nm, for TLC analysis), a UV-Vis spectrophotometer with an associated computer and software, and an LED lamp (315 nm) to irradiate the samples. Combined TLC and UV-Vis spectroscopy allowed the students to identify the extracted ingredients. UV irradiation confirmed the photostability of sunscreens. [ABSTRACT FROM AUTHOR]

Background: The emergence of resistance to commonly used antibiotics by human infections occurred mostly due to their overuse, that prompted individuals to pursue novel and innovative treatments. The phytochemical characteristics, antibacterial activity, and cytotoxicity of MCF7 cells were evaluated in two Pleurotus spp. mycelial extracts in this work. Methods: Pleurotus djamor and Pleurotus florida mycelial extracts from pure cultures were tested for antibacterial activity by a well-diffusion assay and antimicrobial activity against mold fungi was evaluated for biomass inhibition. Mycelial extracts were obtained from dichloromethane extracts and their biophysical characteristics are analyzed by UV–vis spectrum and FTIR analysis. By spraying detection reagents onto TLC plates, the chemicals in dichloromethane extraction of chosen mushroom fungus mycelia were identified. Using the MTT test, the cytotoxic effect of dichloromethane extracts of selected mushroom fungi was evaluated on MCF7 Cell lines. Results: Mycelial extracts of P. djamor and P. florida exhibited significant antimicrobial effect on the bacterial and fungal pathogens tested. Dichloromethane mycelial extracts were obtained using soxhlet extraction which response positive for various phytochemical analysis. Detection of metabolites in thin layer chromatography using spray reagents documented one of few first accounts on flavonoids, anthroquinone and terpenoid compounds in P. djamor and P. florida. P. djamor and P. florida had dose-dependent antiproliferative activity against MCF7 cells, with an inhibitory impact of 55.72% and 64.47% percent at 125 μg/mL, respectively. Conclusion: The study has reported the identification with the potent biological activity of some of the key bioactive components present in DCM extracts from the mycelia of P. djamor and P. florida.

[Display omitted] • Two/four/six units of azobenzene moieties were assembled on the cyclotriphosphazene (CPZ) core. • In solid-state, the N=N functionality in azophenolate units exists in the trans conformation. • The azobenzene moiety in these molecules switched from trans to cis conformation even without UV irradiation. • An inexpensive thin-layer chromatographic technique proves the formation of ZZ isomer in the photoisomerization study. Multi-azobenzene derivatives capable of showing multi-stimuli responsive material can exhibit superior performance in biomedicine, the dye industry, cosmetics, and photonics. Here, a nucleophilic substitution reaction was employed to assemble two (Az 2 CPZ(biph) 2) or four (Az 4 CPZ(biph)) or six (Az 6 CPZ) units of azobenzene moieties on the cyclotriphosphazene (CPZ) core. We have observed that the color of each compound varied by changing the number of azobenzene moieties on the CPZ core. These compounds were characterized by elemental analysis, 1H, 13C{1H}, and 31P{1H} NMR techniques. Single crystal X-ray diffraction techniques ensure the trans conformation in the azobenzene for Az 2 CPZ(biph) 2 and Az 4 CPZ(biph). Electronic spectroscopy for Az 2 CPZ(biph) 2 , Az 4 CPZ(biph) , and Az 6 CPZ having π-π* transition around 325 nm show a hypsochromic shift with ∼25 nm compared to the parent molecule, 4-hydroxyazobenzene AzH. As a result of the electronic spectroscopy, the cis isomer formation enhanced upon increasing the concentration of Az 2 CPZ(biph) 2 / Az 4 CPZ(biph) / Az 6 CPZ in DCM/DMF. The photoisomerization study confirms that these molecules having at least one azo moiety can switch from trans to cis conformation upon exposure to 365 nm light. Interestingly, the thin-layer chromatography (TLC) corroborated the switch of trans to cis conformation occurs at room temperature in a solution state even without UV irradiation. The proton NMR integration provides the generation of the cis -azobenzene moiety is about 12–20 % in Az 2 CPZ(biph) 2 upon exposure to 365 nm light for 15 to 60 mins. Additionally, TLC verifies the generation of multiple isomers that confirm multi-functional switch behavior when the sample with UV exposure time increases. Owing to these molecules undergoing a isomerization process (trans - a z o ⇌ c i s - a z o) even without UV, their fluorescence gets quenched in the solution state. Cyclic voltammetry confirms the quasi-reversible type redox behavior of all compounds in dry dichloromethane. We believe the results presented in this work will be significant for dye and biomedical applications. [ABSTRACT FROM AUTHOR]

In recent years, the interest in natural remedies has increased, so it is important to analyze the plants widely distributed in nature but whose composition is little known. The main objective of the present work is to obtain information based on the profiles of secondary metabolites and antioxidant activity in Lamium album, a very widespread but little studied plant, with the aim of revealing the differences compared to Urtica dioica. First, the optimization of enzymatic extraction assisted by ultrasound was carried out by the Box–Behnken method. The optimized parameters were: concentration of the enzyme—3.3% cellulase, temperature—55 °C, and the extraction time—40.00 min. The efficiency was estimated based on the content of iridoids, the main class of secondary metabolites from Lamium album. Second, the secondary metabolites profiles of the nettle extracts were obtained by thin-layer chromatography using both normal and reverse phases and by RP-UHPLC. The antioxidant activity was evaluated using DPPH and ABTS+ radicals. The obtained results revealed significant differences between the two nettle species, both in terms of the phytochemical compounds, as well as the antioxidant activity, confirming the fact that Lamium album has a high potential to be used in phytomedicine.

In this work, a screen-printing method was developed to create porous particle-based materials as layers with specifically designed shape to produce microfluidics systems. Among several tested binding agents, xanthan gum was found to be an excellent choice for a printing mixture thickener as well as a durable binder for the resulting material. In addition to demonstrating control over the shape of the printed microfluidics chips, control over material thickness, wetting characteristics and general method accuracy were also investigated. The applicability of the introduced method was further demonstrated with a development of an exemplary microfluidics chip for quantitative detection of Fe (III), Ni (II), Cu (II), Cd (II), and Pb (II) from a mixed sample at millimolar levels. The novel approaches demonstrated in this article offer new perspective into creating multiplexed on-site chemical analysis tests.

Abstract Background Nanocolloidal human serum albumin radiolabelled with 99mTc provides a diagnostic radiopharmaceutical for sentinel node lymphoscintigraphy. NanoHSA (Nanotop), a commercially available kit, enables the simple preparation of this radiopharmaceutical via reconstitution with pertechnetate eluted from a generator. Thin-layer chromatography is widely used for determining radiochemical purity in clinical nuclear medicine. Quality control methods recommended by the manufacturer were sometimes reported to yield variable results. Therefore, we proposed and evaluated three alternative thin-layer chromatography methods for the quality control of [99mTc]Tc-NanoHSA from a commercially available kit. Results The radiochemical purity of [99mTc]Tc-NanoHSA determined with all methods was reproducible and met the requirements of the SPC and the European Pharmacopoeia (≥ 95%). Our quality control using iTLC-SG chromatographic paper in methyl ethyl ketone mobile phase identified only free pertechnetate as impurity, resulting in > 99% RCP. The quality control using iTLC-SG in 85% methanol or iTLC-SA in 0.9% NaCl identified an additional small fraction of a hydrophilic impurity, resulting in 95–97% RCP. Glucose was identified as a potential 99mTc-carrying hydrophilic species contributing to hydrophilic impurities. Conclusion Our quality control of [99mTc]Tc-NanoHSA with non-polar mobile phase tended to underestimate the amount of hydrophilic impurities, although without compromising the final quality of the radiopharmaceutical. Alternative TLC methods using aqueous mobile phases enabled a more accurate determination of hydrophilic impurities.

The search for new anti-inflammatory drugs with less side effects requires simple, fast and reliable screening methods. In this context, we have developed a sensitive thin-layer chromatographic (TLC) assay on silica gel plates to detect cyclooxygenase-2 (COX-2) inhibition. COX-2 catalyzes two sequential enzymatic reactions: a first oxygenation step that converts arachidonic acid into prostaglandin G2, and a subsequent reduction of prostaglandin G2 into prostaglandin H2. Our test is based on the co-oxidation during this peroxidation step of a co-substrate, N,N,N′,N′-tetramethyl-p-phenylenediamine (TMPD), leading to a blue-grey product. As a consequence, COX-2 inhibitors appear on the TLC plate after revelation as clear spots against the colored background. Parameters such as concentrations of enzyme, substrate, and chromogenic reagent have been optimized. The limit of detection was found to be below the microgram for standard COX-2 inhibitors such as celecoxib or ibuprofen. The developed TLC assay was also conclusive when applied to 60 various natural pure compounds and some complex natural extracts. Results demonstrated a COX-2 inhibitory activity mostly for triterpene and sterol derivatives. This COX-2 TLC assay appears as a suitable low-cost and reliable strategy for the screening of natural extracts to discover new anti-inflammatory compounds. [ABSTRACT FROM AUTHOR]

This study aimed at optimizing conditions for increased withanolide production in Withania somnifera. The elicitors used for the foliar spray on the aerial parts of the plant were salicylic acid, jasmonic acid, and chitosan for the enhancement of withanolides in Withania somnifera under different environmental regimes. Three different elicitors, i.e., chitosan, jasmonic acid and salicylic acid, were applied on the plants through foliar route every 15th day for 6 months, and later plants were used for sample preparation. Further, the elicitors were used in different concentration, i.e., jasmonic acid (50, 200 and 400 ppm), chitosan (10, 50 and 100 ppm) and salicylic acid (0.5, 1 and 2 ppm). The elicitors were sprayed on the foliar parts of the plant between 10:00–11:00 a.m. on application days. For elicitor spray, a calibrated sprayer was used. The withanolide A/withaferin A was quantified through HPLC. It was found that in an open environment, maximum withaferin A content, i.e., 0.570 mg/g (DW), was recorded with jasmonic acid (50 ppm) treatment in comparison to control (0.067 mg/g DW). Thus, there was an 8.5-fold increase in the withaferin A content. Maximum withanolide A content of 0.352 mg/g (DW) was recorded when chitosan (50 ppm) was sprayed, while in the control, withanolide A content was recorded to be 0.031 mg/g (DW); thus, chitosan application increased the production of withanolide A by 11.3-fold. Under controlled conditions, maximum withaferin A content of 1.659 mg/g (DW) was recorded when plants were sprayed with chitosan (100 ppm), which was 8.1 times greater than the control content of 0.203 mg/g (DW). Maximum withanolide A content of 0.460 mg/g (DW) was recorded when chitosan (100 ppm) was applied, whereas in the control, withanolide A content was found to be 0.061 mg/g (DW). Thus, foliar spraying of elicitors in very low concentrations can serve as a low-cost, eco-friendly, labor-intensive and elegant alternative approach that can be practiced by farmers for the enhancement, consistent production and improved yield of withanolide A/withaferin A. This can be a suitable way to enhance plant productivity, thus increasing the availability of withanolide A and withaferin A for the health and pharma industry. [ABSTRACT FROM AUTHOR]

In our previous paper we have presented the new prototype equipment and introduced a new analytical technique—high-performance/high-pressure layer electrochromatography (HPLEC), a combination of overpressured-layer chromatography (OPLC) and pressurized planar electrochromatography (PPEC). In this paper, the work of the equipment in various operational modes is investigated. Some difficulties and challenges related to various aspects of separation are discussed. The OPLC and HPLEC techniques are compared in terms of selectivity and performance. The results show that our equipment can be successfully used for singe- and multichannel OPLC and HPLEC separations in various sample application and detection modes. It includes the high-throughput, multichannel, and fully automated online separation of multiple samples simultaneously. The equipment allows for the independent optimization of various operational parameters. HPLEC combines the advantages of column/capillary and planar separation techniques while overcoming their limitations. It also combines the advantages and overcomes the drawbacks of OPLC and PPEC. It provides hydrodynamic flow of the mobile phase, irrespective of the voltage used and/or the mobile phase composition. Thus, any optimization of the composition and the voltage can be used independently. Both can be used to obtain the required selectivity of separation. The voltage can be used to facilitate the mobile phase flow and accelerate the analysis. [ABSTRACT FROM AUTHOR]

The advantages of hyperspectral imaging in videodensitometry are presented and discussed with the example of extracts from 70 Polish grasses. An inexpensive microscope camera was modified to cover the infrared spectrum range, and then 11 combinations of illumination (254 nm, 366 nm, white light), together with various filters (no filter, IRCut, UV, cobalt glass, IR pass), were used to register RGB HDR images of the same plate. It was revealed that the resulting 33 channels of information could be compressed into 5–6 principal components and then visualized separately as grayscale images. We also propose a new approach called principal component artificial coloring of images (PCACI). It allows easy classification of chromatographic spots by presenting three PC components as RGB channels, providing vivid spots with artificial colors and visualizing six principal components on two color images. The infrared region brings additional information to the registered data, orthogonal to the other channels and not redundant with photos in the visible region. This is the first published attempt to use a hyperspectral camera in TLC and it can be clearly concluded that such an approach deserves routine use and further attention.

Ambient desorption/ionization mass spectrometry (ADI-MS) is widely used as a rapid screening tool of samples in their native state without sample preparation. While analysis times are much less than 1 min per sample, one challenge of ADI-MS is the possibility to perform quantitative analysis of analytes in complex matrices. Typically, the goal is to probe a variety of different analytes in a complex matrix from a solid, liquid, or otherwise uncharacterized surface in the open air in front of the MS inlet. In this study, it is demonstrated that a carefully selected surface for analyte spot sampling and co-deposited isotopically labeled standards both significantly improve the capabilities of flowing atmospheric-pressure afterglow (FAPA) high-resolution (HR) MS for direct quantitative analysis. Specifically, a systematic study of different surfaces (glass, steel mesh, high-performance thin-layer chromatography (HPTLC) stationary phases including silica, reversed-phase (RP)-modified silica, and cyano (CN)-modified silica) and their suitability for spot sampling with FAPA-MS was performed. A set of different caffeine-containing standards and beverages (Red Bull, Coca-Cola, coffee, and black tea) was deposited on the surfaces and direct FAPA-HR-MS analysis of caffeine was performed using internal calibration with co-deposited 13C3-caffeine. For TLC surfaces, it was demonstrated that quantitative results could be achieved with the matrix and concomitants present and that a preceding chromatographic separation was not mandatory for this application. In addition, the use of a CN-HPTLC surface resulted in a significantly more intense caffeine signal in the beverage samples compared to the other surfaces studied, with the highest increase compared to the silica (200-fold higher) and the lowest increase compared to the steel mesh (30-fold higher). The utilization of TLC-based surfaces as sample carriers is considered an attractive tool in the ADI-MS toolbox for fast and efficient mass spectrometric investigations of complex samples without time-consuming sample preparation. [ABSTRACT FROM AUTHOR]

In the alarming context of antibiotic resistance, we explored the antibacterial potential of French mushrooms against wild-type and multidrug-resistant (MDR) bacteria. In order to accelerate the discovery of promising compounds, screenings were carried out by TLC-direct bioautography. A total of 70 extracts from 31 mushroom species were evaluated against five wild-type bacteria: Staphylococcus epidermidis, Staphylococcus aureus, Enterococcus faecalis, Escherichia coli, and Pseudomonas aeruginosa. This first screening revealed that 95% of the extracts contained antibacterial compounds. Generally, it was observed that EtOAc extracts exhibited more active compounds than methanolic extracts. In addition, all extracts were overall more active against Gram-positive bacteria than against Gram-negative strains. The most promising mushroom extracts were then screened against various multidrug-resistant strains of S. aureus and E. coli. Activity was globally less on MDR strains; however, two mushroom species, Fomitopsis pinicola and Scleroderma citrinum, still contained several compounds inhibiting the growth of these MDR pathogenic bacteria. Stearic acid was identified as a ubiquitous compound contributing to the antibacterial defence of mushrooms. This screening revealed the potential of macromycetes as a source of antibacterial compounds; further assays are necessary to consider fungal compounds as promising drugs to counter antibiotic resistance. [ABSTRACT FROM AUTHOR]