Chromatographic Analysis Research Articles

Impact of Paracoccus sp. EGY7 carotenoids on triple-negative breast cancer cells: invitro study

This study investigates the effect of Paracoccus sp. EGY7 carotenoids on the triple-negative breast cancer cell line (MDA-MB-231). The bacterial strain was isolated, and its carotenoids profile was analyzed via HPLC-DAD-MS. Cytotoxicity, migration tests and the expression of BAX and BCL-2 at the gene and protein levels were carried out to evaluate the therapeutic efficiency of the produced carotenoids. Molecular docking analysis estimated the binding affinity between zeaxanthin and BCL-2 protein. Chromatographic analysis revealed zeaxanthin as the major carotenoid (48.3%). The extract exhibited significant cytotoxicity against MDA-MB-231 cells with an IC50 of 1200 µg. It notably reduced cell migration, with wound closure percentages of 37.50% and 79.17% for the 600 µg group, while the percentages were 12.50% and 53.50% for the 1200 µg group, compared to 71.67% and 95.67% for the control at 24 and 48 h post-treatment, respectively. The extract induced apoptosis, as evidenced by significantly increased BAX/BCL-2 gene expression ratios at 600 and 1200 µg (p < 0.05). Western blotting showed increased BAX protein expression at 600 and 1200 µg compared to the control group (p < 0.001), and significantly lower BCL-2 protein expression (p = 0.000005 for 1200 µg and p = 0.0001 for 600 µg). Docking analysis indicated a strong affinity of zeaxanthin to BCL-2 (ΔG = -9.773241 kcal/mol) compared to obatoclax (ΔG = -7.419345 kcal/mol). Paracoccus sp. EGY7 carotenoids are a promising anticancer agent against MDA-MB-231 cells. They effectively promote apoptosis and prevent metastasis, crucial for disease advancement in cancer cells.

Retinoids in lipofuscin granules from retinal pigment epithelium as biomarkers of the damaging effect of ionizing radiation.

Lipofuscin granules accumulate in the retinal pigment epithelium with age, especially in patients with visual diseases, including progressive age-related macular degeneration. Retinoids (bisretinoids and their oxidation products) are major sources of lipofuscin granule fluorescence. The aim of this work was to analyze the radiation-mediated oxidation of retinoids in lipofuscin granules obtained from the human cadaver eye retinal pigment epithelium. Fluorescent and chromatographic analyses of retinoids were performed before and after irradiation of lipofuscin granules with accelerated protons. The fluorescent properties of chloroform extracts from irradiated lipofuscin granules exhibited an increase in fluorescence intensity in the short-wavelength region of 555 nm. This change is associated with an increase in the quantity of retinoid oxidation cytotoxic products after accelerated proton exposure. The radiation-induced oxidation of retinoids caused a noticeable change in the fluorescent properties of retinoids allows us to consider this phenomenon as a potential opportunity for non-invasively assessment of the degree of radiation exposure and its relative biological effect in humans. Thus, this research proposes a new strategy for assessing the extent of radiation exposure to humans, which evaluates the effects of ionizing radiation on human eye tissues. This approach is based on the principles of the modern non-invasive method of fundus autofluorescence used in ophthalmology for the diagnosis of the retina and retinal pigment epithelium degenerative diseases.

Blue applicability grade index assessment and response surface modelling to synchronous determination of metformin hydrochloride, vildagliptin and dapagliflozin propanediol monohydrate by HPTLC method

Diabetes mellitus is major chronic disease found in human due to stressful lifestyle and bad food habit. Recently, the Central Drugs Standard Control Organization of India has approved combination of metformin hydrochloride, vildagliptin and dapagliflozin propanediol monohydrate for the management of diabetes mellitus. Numerous chromatographic methods have been published in the literature for estimation of these anti-diabetic drugs on individual basis and their combinations. But these chromatographic analyses promoted usage of toxic organic solvents which are harmful to the environment and aquatic animal lives. In addition, no chromatographic method was found in the literature for simultaneous estimation of these anti-diabetic drugs using green solvents. In recent days, white analytical chemistry approach has been introduced in the literature for development and assessment of green, accurate, precise, cost-effective and user-friendly chromatographic methods. Hence, white analytical chemistry-driven risk-based HPTLC method was developed for synchronous estimation of these anti-diabetic drugs using green solvents and response surface modelling. The analytical quality risk assessment was applied for identification of critical method variables and response variables using risk priority number ranking and filtering method. The critical method variables were linked with critical response variables using response surface modelling by Box–Behnken design. The analytical design space was navigated control strategy was framed for robust densitometric estimation of anti-diabetic drugs. The chromatographic method was validated as per ICH Q2 (R2) guideline. The developed method was applied for synchronous assay of multiple combinations of metformin, vildagliptin and dapagliflozin using single chromatographic condition to save time, cost and resources for analysis. The assay results of these anti-diabetic drug combinations were found to be complied with the respective labelled claim of the drugs. The developed and published chromatographic methods were assessed for their redness, greenness, blueness and whiteness profiles using green analytical metrics and RGB model scoring system. The proposed method was found to be green, cost-effective, robust and user-friendly for synchronous estimation of metformin, vildagliptin and dapagliflozin.

A simple HPLC-UV method for monitoring therapeutic adherence in pulmonary arterial hypertension.

A considerable percentage of ineffective treatment in pulmonary arterial hypertension (PAH) may be related to subtherapeutic dosage or non-adherence. The aim of the study was to develop a simple analytical method suitable for plasma determination of selected drugs: riociguat (RIO), bosentan (BOS) and macitentan (MAC) administered to PAH patients. An isocratic HPLC-UV system (Spectra Physics - Shimadzu) with a manual injector (50μL loop) was applied. Chromatographic analysis was performed using a Suplecosil LC-CN column (150×4.6mm, 5μm) protected with a Supelguard precolumn at room temperature. The separation was carried out using the mobile phase: CH3CN:H2O:0.5M KH2PO4:85% H3PO4 (172:324.2:3.7:0.1, v/v) at a flow rate of 1.8mL/min. Ethyl acetate (4mL) was used for 10-min liquid-liquid extraction from 0.4mL alkalized plasma sample. Detection was performed at λ=245nm chosen as a compromise between signal intensity and matrix interference. The analytes were eluted at retention times of 4.4min (RIO), 5.4min (BOS), 8.9min (MAC) and 7.8min for gallopamil (internal standard, GAL). The method was found linear and calibrated in the ranges: 5-1000ng/mL for RIO, 10-2000ng/mL for BOS and 20-2000ng/mL for MAC, with r2 of 0.9991 for RIO, 0.9983 for BOS, and 0.9949 for MAC, respectively. Within the given ranges, the method ensured reliable results with the required precision and accuracy: ≤15% (≤20% for LLOQ). There was no significant carryover effect. The method has been successfully used in pilot study on adherence in patients treated for PAH, enabling monitoring of RIO, BOS and MAC. Drug concentrations were assessed in samples taken before (C0) and 3h after drug administration (C3). For RIO, BOS and MAC, the developed method was suitable for both C0 and C3 samples, allowing steady-state drug determination if used. The presented method can be recommended to laboratories equipped with basic HPLC apparatus as an attractive analytical tool for both TDM and adherence studies.

Research advance of solid-phase microextraction based on covalent organic framework materials

Solid-phase microextraction (SPME) is a fast and simple sample preparation technique that enables the enrichment of analytes, and it is used in combination with other detection techniques to provide accurate and sensitive analytical methods. SPME is widely used in environmental monitoring, food safety, life analysis, biomedicine, and other applications. The extractive coating is the core of the SPME technique, and the properties of the extractive coating greatly influence extraction selectivity and efficiency, as well as the enrichment effect. Therefore, the development of new and efficient extractive coating materials remains a hot topic in the analytical chemistry and sample preparation fields. Covalent organic frameworks (COFs) are a kind of porous crystalline network polymer materials formed by covalent bonds. Owing to the advantages of large specific surface area, high porosity, good stability, high designability, simple synthesis and post-modification, etc., it has been widely used in gas adsorption, catalysis, sensing and drug delivery. In recent years, COFs have attracted much attention in the field of sample preparation. A variety of novel COF-based SPME materials had been developed for extracting and enriching various types of analytes through π-π interaction, hydrophilic/hydrophobic interaction, electrostatic adsorption, and hydrogen-bonding, as well as pore effects. In this paper, the research advances of COFs for using in fiber-, in tube-, and membrane-based SPME over the past three years were discussed. Fiber surfaces had been modified with functionalized COFs or COF-hybrid materials for use in SPME through physical coating, in-situ growth, and chemical-bonding approaches. The combination of SPME fiber and chromatographic analysis can be used to detect a variety of analytes such as polycyclic aromatic hydrocarbons, phthalates, polychlorinated biphenyls, and pesticides in environmental and food samples, with good enrichment effects, wide linear ranges, and high sensitivity. Based on in tube-SPME, COFs-based monolithic column and fiber-filled tube as the extraction tubes were combined with high performance liquid chromatography online to develop highly sensitive detection methods for synthetic phenolic antioxidants and bisphenol compounds, respectively. In addition, COFs had also been used in membrane SPME technique, showing high efficiency in the extraction of trace polychlorinated biphenyls in environmental water. Finally, the development trends of COFs in the field of SPME was prospected.

Purification of mineral oil aromatic hydrocarbons and separation based on the number of aromatic rings using a liquid chromatography silica column. An alternative to epoxidation.

The analysis of mineral oil aromatic hydrocarbons (MOAH) in vegetable oils is currently associated with high uncertainty due to various factors ranging from sample preparation to data interpretation. One significant factor is the coelution of biogenic compounds of terpenic origin with the MOAH fraction during chromatographic analysis. The common purification method is epoxidation, a chemical reaction that changes the polarity of the interferences, allowing their separation from MOAH. However, this reaction is non-selective and can lead to losses of both MOAH and internal standards. This variation is especially noticeable when using epoxidation procedures with higher reaction kinetics, such as those employing performic acid. MOAH losses also vary depending on their composition, which is unpredictable. Furthermore, 2-methylnaphthalene (2MN) and 1,3,5-tri‑tert-butylbenzene (TBB), which are the common quantification standards, are lost at different rates. As a consequence, the final MOAH quantity will vary both depending on the standard used, and on its initial composition. This work presents a new purification approach based on liquid chromatography fractionation on silica using the same column and eluents as for the usual MOSH/MOAH fractionation. This approach efficiently removes squalene, carotenes, and derivatives, without inducing inconsistent losses between the internal standards and MOAH as epoxidation does. On average, MOAH recovery using this new method was 94% (± 8%) in coconut, palm, sunflower, and olive oil, using different MOAH sources and concentrations. Additionally, perspectives are presented regarding the separate analysis and quantification of mono-/diaromatic MOAH and other polyaromatic MOAH. This is of particular interest as these two sub-fractions are associated with different toxicological properties.

Comparison of major carotenoid composition in palm fruits (Arecaceae): An analysis of oil palm fruits and the colored exocarp fruits of four palm species.

The oil palm, a widely studied species of palm, is a crucial source of edible oil and pro-vitamin A carotenoids, primarily α- and β-carotene. The diverse peel and pulp colors (yellow-orange-red) of various palm fruits suggest the presence of not only pro-vitamin A carotenoids but also other bioactive carotenoids like lutein and lycopene, which offer additional health benefits. This study aimed to identify and quantify the major carotenoids in fruits with colored exocarps of four varieties of palm species belonging to different genera and compare them to those in oil palm fruit to evaluate their bioactive potential for human health. High-performance liquid chromatography and mass spectrometry were used for chromatographic and spectrometric analyses using non-saponified samples, except for lutein estimation, revealing four major carotenoids: α- and β-carotene, lycopene, and lutein, which together accounted for more than 50% of the total carotenoid content. Among the five palm fruits tested (MacArthur, Manila, mountain date, foxtail, and oil palms), foxtail palm exhibited the highest concentration of major carotenoids at 500µg/g dry weight (d.w.), with β-carotene comprising 69% of this total, followed by MacArthur palm (235µg/g d.w.), whereas mountain date palm had the lowest concentration of major carotenoids at 135µg/g d.w. These findings suggest that colored palm fruits are significant sources of both pro-vitamin A carotenoids and other beneficial carotenoids like lycopene and lutein. The method of sample preparation, particularly the inclusion or omission of a saponification process, plays a critical role in the recovery and accurate quantification of carotenoid concentrations due to their varying susceptibility to this process.

Global retention model based on multisample system parameters for optimising chromatographic fingerprints of medicinal plants.

Developing analytical methods for Traditional Medicine products by liquid chromatography is challenging due to their chemical complexity and the lack of analytical standards for numerous, unidentified constituents. Regulatory agencies recommend chromatographic fingerprint analysis for quality evaluation, relying on peak detection to ensure resolution. Conventional modelling struggles to optimise experimental conditions for such complex samples. Recent research highlights that global models, which integrate column/solvent, and solute-specific parameters, effectively streamline method development. This work demonstrates the feasibility of characterising separation systems using chromatograms from independent experimental designs across multiple plants (lemon balm, green tea, and linden), which are processed altogether to build global models. The multisample system parameters enable the optimisation of gradient programs, not only for the three plants used to develop the generalised global model, but also for external plants (peppermint and pennyroyal), with the acquisition of only an additional single gradient experiment. The MATLAB Global Optimisation Toolbox is tested and compared with the previously applied alternating regression method, providing a convenient and accessible solution for fitting global models. The agreement between experimental and predicted optimal separations found with global models is comparable to the accuracy achieved with models specifically tailored to each solute. This research demonstrates the benefits of enhancing global models with data from multiple plant species, enabling robust characterisation of column and solvent properties. The generalised models allow transferable system parameters and accurate predictions for new plants with minimal experiments. This approach effectively separates solute retention effects from column and solvent influences, achieving predictive capabilities comparable to traditional individual retention models, without requiring standards for each compound.

Evaluation of the Performance of Microalgae and Constructed Wetlands for the Reduction of Toxicity Organic Load and Pharmaceuticals from Wastewaters

The present study aimed to evaluate the performance of a novel integrated system composed of microalgae and constructed wetlands (CW). The microalgae production tank was designed as a raceway type, and microalgae of the genus Chlorella were inoculated, while the CW was established as the first stage of the French model and vegetated with the macrophytes Chrysopogon zizanioides, Typha domingensis, and Dracaena trifaciata. The hydraulic retention time in each unit was 7 days, totaling 14 days of treatment. Good reductions in dissolved organic carbon (67.2%) and Total N (68.6%) were achieved after the treatment. The treatment also completely eliminated acute ecotoxicity against Daphnia magna and genotoxicity in the Allium cepa test assay. Liquid chromatographic analysis revealed the presence of 9 pharmaceuticals in the raw wastewaters, whereas only residual dipyrone was detected after the CW unit. Thus, the novel system investigated in our study, which combines microalgae and CW using 3 different species of macrophytes, proved to be a promising alternative for treating urban wastewaters, as it effectively reduced organic load, nutrient content, toxicity, and removed emerging contaminants such as pharmaceuticals. Future recommendations include investigating the primary removal mechanisms of the pharmaceuticals and enhancing the separation process of the microalgae.

Design and evaluation of a liquid chromatographic column oven with adaptive and precise temperature control

High performance liquid chromatography (HPLC) is a key analytical technique that is used in a number of fields. Improving the separation efficiency, stability, and universality of HPLC has been a continuing analytical-chemistry focus. In chromatographic separation, factors such as the composition and ratio of the mobile phase, the type of stationary phase, and the dimensions of the chromatographic column significantly affect the separation efficiency. In addition, the temperatures of the chromatographic column and mobile phase are also important for achieving separation. The column oven is usually used to stably control the column temperature in the HPLC separation system. Indeed, highly accurate temperature control ensures superior separation performance, short analysis times, and repeatability. In this study, we innovatively improved the traditional column oven by combining a variety of temperature-control algorithms to deliver continuous and highly accurate temperature control in the wide 4-90 ℃ range, and by exploring a new chromatographic-method development route. Instead of focusing on the complex hardware system, we optimized the software to achieve highly stable and accurate (±0.1 ℃) temperature control. Temperature-control performance was further improved by optimizing the structure of the thermal insulation and employing reliable and environmentally friendly thermal-insulation materials. Additionally, the thermal conduction of the heat-source device is discussed based on the heat-transfer principle with the aim of improving the performance of the column oven. The improved column oven delivered significantly enhanced chromatographic-separation repeatability and stability thereby reliably guaranteeing the development of highly efficient chromatographic analysis methods.

BIOREMEDIATION OF PETROLEUM REFINERY EFFLUENTS USING MUTATED FUNGAL CONSORTIA: A CASE STUDY FROM KADUNA, NIGERIA

Bioremediation is a process of using naturally occurring species to break down hazardous substances into less harmful or non-toxic substances. Effluents from petroleum refinery pose a serious health hazard on the people who depend on the water as source of supply for domestic uses. To avoid health hazard, it is imperative for these toxic compounds to be removed from waste water before its disposal. The primary goal of this study was to carry out bioremediation of petroleum refinery effluents using mutant strains of fungal consortia. Refinery effluents were collected under the storage tank in Kaduna Refining and Petrochemical Company, Kaduna, Nigeria. Effluent samples were analyzed for total petroleum hydrocarbons using Gas Chromatography and Mass Spectroscopy (GC-MS). Standardization of the fungal consortium inoculum was prepared following standard method. The culture medium of each isolate of the mutated fungi Aspergillus niger, A. fumigatus, A. versicolor and A. quadrilineatus was carried out following standard procedure. The result of the GC-MS analysis revealed that cis-13-Octadecenoic acid, methyl ester; 7-Hexadecenoic acid, methyl ester; 10-Octadecenoic acid, and Methyl stearate; showed the highest percentage quality of (99%) respectively. Bioremediation of Kaduna refinery effluents in this study indicated that, there was decline in the bioremediation parameters in the second week up to the fourth week whereby the COD, BOD all showed significant decrease. Based on the Gas Chromatographic (GC) analysis of the fungi consortium, the degradative ability of the mutants of Aspergillus versicolor and Aspergillus quadrilineatus were observed to be prominent.

Chromatographic Analysis of Polyherbal Extract and Formulation by HPTLC and GC-MS Methods

Background: The World Health Organization (WHO) acknowledges that natural plants and their bioactive components have been recognized for their potential therapeutic benefits. The main issue facing the herbal industry is the lack of effective quality control for herbal extracts and formulations Establishment standards are essential to ensuring the efficacy and integrity of natural medicine development. Determining the precise compound responsible for therapeutic action poses an equally challenging task, as these compounds frequently work synergistically to produce medicinal benefits. Aim: In this exploration, GC-MS and high-performance thin-layer chromatography were used to analyze the formulation and polyherb extract. The HPTLC technique is often employed to analyze the sample and compare its fingerprint to reference standards to verify plant extracts’ quality and formulations’ quality. Using GC-MS, the phytochemical ingredients in the formulation and the extract have been determined. Methods: This research assessed a mixture of herb extract and formulation using analytical approaches. The combined hydroalcoholic extracts of the herbs were used to produce the polyherb extract of seeds of Azadirachta indica, Carica papaya, Trigonella foenum-graecum, Gossipium herbaceum, fruit of Piper nigrum. The required excipients were added before compressing the herbal mixture extract into tablets. Results: The extract and formulation’s HPTLC chromatograms displayed Rf values of 0.14, 0.34, 0.48, 0.63, and 0.76.The results of the GC-MS, N-Hexadecanoic, 1,19-Eicosadiene, 9,12-Octadecadienoic Acid, Z, Z-4,16-Octadecadien-1-Ol Acetate, Piperine that were analyzed provide different peaks to determining the presence of phytochemicals found in the extract and formulation were confirmed. Conclusion: HPTLC and GC-MS aid in the identification of individual chemical constituents within herbal formulations by coupling chromatographic and mass spectral data of the sample with reference standards or databases, the identity of key compounds can be confirmed, ensuring authenticity, and has shown an intense correlation between the mixture of herb extract and formulation. Major Findings: In this Research, GC-MS and high-performance thin-layer chromatography were used to analyze the formulation and polyherbal extract of key compounds can be confirmed, ensuring authenticity, and has shown an intense correlation between the mixture of herb extract and formulation.

Fixed Oils of Medicinal Palms of Arecaceae from Chapada do Araripe: Chemical Composition and Antibacterial Potential

This study aimed to analyze the chemical composition of fixed oils extracted from the fruits of Acrocomia aculeata, Syagrus cearensis and Attalea speciosa, in addition to evaluating their efficacy in combating resistant microorganisms, such as Escherichia coli and Staphylococcus aureus. The ripe fruits were collected in the region of Barbalha, Ceara , and the extracted oils were analyzed by gas chromatography and mass spectrometry (GC/MS) to identify the compounds present. The antibacterial activity was tested using the microdilution method in 96-well plates, evaluating the inhibition of bacterial growth at different concentrations of the oils. The chromatographic analysis of the fixed oils of Acrocomia aculeata, Attalea speciosa and Syagrus cearensis revealed the predominance of saturated fatty acids, with lauric acid being the most abundant (41.71% to 47.21%). Oleic and myristic acids were also significant, while stearic and linoleic acids appeared in smaller amounts. Attalea speciosa showed inhibition of 40.17% against Escherichia coli and 40.77% against Staphylococcus aureus (1000 μg/mL). Acrocomia aculeata inhibited 44.76% of S. aureus (1000 μg/mL), and Syagrus cearensis had moderate activity against E. coli.

In-depth study of the nutritional composition, phytochemicals, antioxidant activity, molecular docking interactions, and toxicological evaluation of Abies marocana Trab. woody biomass

IntroductionMorocco is renowned for its rich plant biodiversity, although many plants are underutilized. Consequently, the present study was conducted to assess the nutritional composition, bioactive constituents, antioxidant properties, and acute oral toxicity of Abies marocana's woody biomass.MethodsThe nutritional value of the twigs and cones was determined using the AOAC method, while mineral components were examined through ICP-OES. To search for phytochemicals in the methanolic extracts, a number of established techniques were applied, and evaluate their antioxidant activity, while the volatile content was determined using the GC-MS method. The acute oral toxicity test was carried out following the guidelines of OECD 423.Results and discussionProximate analysis revealed a variety of components in different parts, including moisture, ash, fiber, protein, carbohydrates, and fat. Cones and twigs were found to be rich in mineral elements, as well as phenols, flavonoids, tannins, and phytosterols. Chromatographic analysis identified linoleic acid as the main component in twig extract and 2-Bornanone as the primary constituent in cone extract. The methanolic extracts of A. marocana displayed strong antioxidant properties through notable DPPH scavenging activity, with no mortality observed in rats even at doses exceeding 1,000 mg.kg−1, indicating potential for medicinal, cosmetic, or dietary uses. A molecular docking study of the five main compounds of both plants showed that they aligned and interacted with the binding sites of selected proteins, demonstrating significant antioxidant activity.

P1341 Probing the interaction of probiotic bacterial extracellular vesicles with primary colonic epithelium using an ex-vivo organoid platform

Abstract Background Bacterial extracellular vesicles (BEV) are increasingly recognized as a relevant mode of contactless host-microbiota communication. BEV are nanoparticles pinched off from the membrane of multiple bacteria and carry nucleic acid snippets and functional proteins inherited from their parental bacteria. While evidence suggests that BEV act as transmitters to modulate the immune response, only scarce data exist as to whether and how efficiently BEV are internalized by colonic epithelium as the host‘s front line in the gut. This study aims to quantify BEV internalization into primary colonic epithelium and explore structural and physicochemical properties of BEV potentially influencing uptake. Methods BEV were isolated from Limosilactobacillus reuteri (LR) and Lactiplantibacillus plantarum (LP) and incubated with either colonoids or colonic epithelial monolayers from C57BL/6 mice. BEV were characterized by transmission electron microscopy (TEM), nanoparticle tracking analysis (NTA), dynamic light scattering (DLS), zeta potential (ZP) analysis, as well as lipidomic and proteomic analysis. Uptake of BEV into colonic epithelium was assessed by confocal microscopy and flow cytometry. Results BEV of either species displayed a uniform appearance with a lipid bilayer on TEM. NTA showed similar spectra of sizes with a dominant peak around 100 nm whereas with DLS particle size readings were higher and showed a bimodal distribution. The ZP of BEV measured in buffer with physiological salinity did not differ between LR and LP. Gas chromatographic mass spectrometry analysis revealed neither qualitative nor quantitative differences in the composition of long chain fatty acid in BEV membranes. As opposed to LP, LR pellets showed proteomic enrichment of pathways linked to glycerolipid metabolism, fatty acid biosynthesis, and terpenoid backbone synthesis, being precursor steps of lipoteichoic acid synthesis (LTA). LTA could be detected in BEV of both species, but appeared to more abundant in LR-BEV. A dose-dependent increment in BEV uptake rates was noted in both species, with LR-BEV being more readily internalized by colonic epithelium. Epithelial cells exposed to LR-BEV were characterized by irregular nuclei and partial detachment from the surface whilst cells incubated with LP-BEV appeared morphologically normal. The quantity of viable cells was reduced in colonoids treated with LR-BEV, but not LP-BEV. In the presence of LR-BEV, colonoids were less likely to undergo apoptosis. Conclusion LR- and LP-BEV differ in their ability to enter colonic epithelium and interfere with cellular homeostasis. A variable degree of LTA decoration of BEV membranes might account for differential uptake into the host cell.

Chromatographic and Spectroscopic Analyses of Cannabinoids: A Narrative Review Focused on Cannabis Herbs and Oily Products

Cannabis sativa L. is cultivated nowadays for agricultural, industrial, and medicinal applications and also for recreational use. The latter is due to the presence of delta-9-tetrahydrocannabinol, a psychoactive substance. Recreational cannabis policies vary between different countries, which has led to the lack of a clearly defined legal context for cannabis and also a diversity of products derived from or containing cannabis on the (il)legal market. These cannabis-derived products have regained attention, notably because of their cannabinoid content. This review aims to assess and present analytical methods developed to analyze phytocannabinoids with spectroscopic and chromatographic techniques in specific cannabis matrices: herbs and oily products. Published papers from 2018–November 2024 were searched for with precise criteria, analyzed, and summarized. In the studies, liquid and gas chromatographic techniques (&gt;70% reviewed papers) were the most used and have been widely applied using similar methods, and most papers were focused on cannabis herbs (&gt;75%). Techniques were also compared and future challenges were identified. A comparison of different specificities of chromatographic and spectroscopic techniques discussed in this current review has also been established and summarized.

Biotransformation of Ganoderma lucidum and Hericium erinaceus for ex vivo gut-brain axis modulation and mood-related outcomes in humans: CREB/BDNF signaling and microbiota-driven synergies.

The human gut microbiota plays a crucial role in various aspects of health, extending beyond digestion and nutrient absorption. Ganoderma lucidum (Reishi) and Hericium erinaceus (Lion's Mane), traditional medicinal mushrooms, have garnered interest due to their potential to exert positive health effects. The aim of our study was to investigate the molecular impact of Reishi and Lion's Mane on mood regulation through the gut-brain axis. We utilized a dynamic simulator of the human intestinal microbial ecosystem (SHIME), followed by HPLC-ESI-QTOF-MS/MS and a series of biochemical and molecular assays, including MTT for cell viability, fluorogenic probes for redox balance (ROS and GSH), and Western blot for protein analysis. Chromatographic analysis confirmed the presence of bioactive compounds in both mushrooms, including triterpenoids (ganoderic acids) and polysaccharides in G. lucidum, as well as hericenones and erinacines in H. erinaceus. We observed concentration-dependent changes in metabolic activity and redox balance due to microbiome cell-free supernatant treatment (M-CFSs). M-CFSs also influenced the Nrf2 pathway and activated heat shock proteins, which may confer neuroprotective effects. Notably, M-CFSs upregulated neurotrophic factors such as BDNF, CDNF, and MANF, crucial for neuronal function. Our study revealed alterations in intracellular signaling cascades, most notably the CREB/BDNF pathway. Moreover, the Akt/mTOR and ERK1/2 showed no significant changes, while Akt/GSK3α/β displayed only partial modifications. The overlapping effects of synaptic activity and activation of the gut-brain axis appear to contribute to mood enhancement. These pilot findings suggest a potential role for G. lucidum and H. erinaceus in mood disorder regulation through multifaceted mechanisms involving the gut microbiota. The study underscores the importance of understanding the synergistic interactions between medicinal fungi, gut microbiota, and neural processes to develop novel or preventive strategies for mental health disorders.

Catalytic transesterification of rapeseed oil under supercritical fluid conditions and physical properties of the reaction product

This research examines the transesterification reaction of rapeseed oil with ethanol in the presence of heterogeneous catalysts - Al2O3 and CaO/Al2O3 with 2% mass impregnation of CaO. The experiments were carried out at a molar ratio of ethanol: rapeseed oil of 18:1 (volume ratio ≈ 1:1) at temperatures of 608, 623 K, and a pressure of 20 MPa for up to 30 minutes in a flow-type unit. The state’s selected pressure and temperature parameters correspond to the supercritical fluid conditions. The results are presented for kinematic viscosity (T=313 K, P=0.1 MPa), density (T=288÷323 K, P=0.1 MPa), refractive index (T=298 K, P=0.1 MPa) of the transesterification reaction product. A quantitative assessment of the target product was carried out using chromatographic analysis. We have established the influence of temperature and duration of the reaction process on its efficiency. We present the strength characteristics of the catalysts used before and after the experiments. During attrition, the initial Al2O3 catalyst lost weight by 0.4 ± 0.1%. The mechanical crushing strength along the generatrix was 7.1 ± 0.3 N/mm. The strength of catalysts after carrying out the transesterification reaction under supercritical fluid conditions decreases, on average, by 24-28%. The use of heterogeneous catalysts Al2O3 and CaO/Al2O3 with 2%wt. impregnation of CaO showed an increase in the fatty acid ethyl esters concentration in the transesterification reaction product compared to the reaction carried out without the use of catalysts.

Insight into the Characteristic Components of Cigar Tobacco.

As an important agricultural product, cigar tobacco lacks reports on chemical compositions. The investigation on characteristic components of Yunnan local cigar tobacco revealed 15 Cembrane-type diterpenes including nine undescribed ones (1-9). Their structures were elucidated on the basis of extensive spectroscopic analysis, computational calculations, and X-ray diffraction. The skeleton of Cigarcembrane (1) with a new 6/12-ring system was previously undescribed. This is the first time to report eunicellane (2-7) and gersemiane (8-9) skeletons from plant resources. The transformation mechanism of cigarcembrane, eunicellane, and gersemiane from Cembrane during cigar fermentation was postulated. Chromatographic analysis revealed 10 as a marker for cigars. All the isolated compounds showed the promotion of hOMF cell proliferation, enlightening us to seek lead compounds in mouth care.

Application of Κ-Carrageenan for One-Pot Synthesis of Hybrids of Natural Curcumin with Iron and Copper: Stability Analysis and Application in Papilloscopy

In this study, hybrid materials were synthesized incorporating curcumin, Cu2+ or Fe3+, and Kappa-carrageenan as a reducing agent to improve stability, considering that curcumin has low thermal and solution stability, which limits its applications. Colorimetric analysis showed color changes in the hybrids, ultraviolet–visible spectroscopy revealed band shifts in the hybrids, and infrared analysis indicated shifts in wavenumbers, suggesting changes in the vibrational state of curcumin after bonding with metal ions. These techniques confirmed the formation of hybrid materials. Thermogravimetric and chromatographic analyses demonstrated greater thermal and solution stability for the hybrids compared to curcumin. Additionally, the hybrid composites effectively developed natural and sebaceous latent fingerprints with good clarity and contrast on glass surfaces. Both composites performed similarly to commercial Gold® powder. When applied to surfaces representative of forensic scenarios, the composites were versatile, revealing sufficient fingerprint details for human identification on both porous and non-porous surfaces. Scanning electron microscopy images showed greater clarity in sebaceous and natural fingerprints developed with the Fe composite compared to the Cu composite.