Search Results (148)

Fish are continuously exposed to stress factors throughout their life cycle, making the use of anaesthetics essential for a wide range of experimental procedures. Currently, the most commonly used and FDA approved anaesthetic for fish research is Tricaine Methanesulfonate (MS-222). However, its use has been associated with several undesirable effects, including hypoxemia, hypercapnia and hypoglycaemia, as well as environmental concerns due to its release through aquaculture effluents. These limitations highlight the need for alternative anaesthetic strategies. Natural compounds such as clove oil, menthol and thymol have been investigated as potential alternatives, demonstrating effective anaesthetic properties. However, their low aqueous solubility, represents a significant challenge, which may be overcome through nanoencapsulation. This approach can enhance solubility, enable controlled release, and reduce the effective dose required. Accordingly, the present study aims to provide an overview of the recent advances in nanoparticle-based encapsulation strategies for anaesthetic delivery in fish, with a focus on their efficacy, safety and environmental impact. Some studies have demonstrated the benefits of nanoencapsulation. In adult zebrafish (Danio rerio), lower concentrations of benzocaine were required when encapsulated in chitosan-PLGA nanoparticles, while lidocaine-loaded lipid NPs reduced bradycardia. In Nile Tilapia (Oreochromis niloticus), clove oil encapsulated in lipid-based nanocapsules enabled effective anaesthesia and prolonged release of the active compound eugenol. Similarly, mucoadhesive zein NPs, reduced the effective concentration of Eugenol by up to 50%. Monoterpenes such as menthol and thymol also show promise for zebrafish anaesthesia, demonstrating efficacy at 50 mg/L. These findings suggest that nanoparticle-based delivery systems can improve the efficacy and safety of fish anaesthetics while reducing required doses and potential environmental impact. Future research should focus on optimizing nanoparticle-anaesthetic systems by combining natural compounds with biocompatible and biodegradable nanocarriers (e.g., zein, chitosan or PLGA) to achieve controlled release, targeted delivery and minimization of side effects. Full article

Essential oils (EOs) from wild Calamintha nepeta (Portugal) populations collected in Portugal (Évora) were investigated in order to evaluate the impact of Mediterranean seasonal conditions on their phytochemical composition and biological activity. Essential oil GC-FID and GC-MS analyses revealed distinct seasonal chemotypes, with spring samples dominated by isopulegone/pulegone, whereas autumn samples contained higher proportions of isomenthone and menthol. Antioxidant activity was assessed through lipid peroxidation inhibition, DPPH radical scavenging and ferric reducing power assays, while antibacterial activity was evaluated against multidrug-resistant (MDR) clinical isolates. Seasonal differences were reflected in both EO chemical composition and bioactivity. Autumn samples displayed greater antioxidant potential, with Y1A showing the highest inhibition of lipid peroxidation (IC₅₀ = 0.85 mg/mL) and Y2A exhibiting the highest ferric reducing power. Conversely, spring samples were more active against MDR bacteria. Among them, Y1S showed the broadest antimicrobial spectrum, with MIC values ranging from 465 to 1767 μg/mL. The unusually wet spring season coincided with marked isopulegone accumulation (≈50%), while warmer autumn conditions favoured higher levels of isomenthone and menthol in the EOs. These findings highlight the importance of seasonal environmental conditions in determining the phytochemical profile and bioactive potential of C. nepeta EOs, providing valuable insights for their standardisation and valorisation in pharmaceutical, food and conservation-related applications. Full article

Deep eutectic solvent (DES) extraction is a green and efficient technology. As a substitute for organic reagents, DESs are widely used to extract active ingredients from traditional Chinese medicine. This study established an environmentally friendly and efficient method for extracting astaxanthin (AST) from Phaffia rhodozyma (PR) using ultrasound-assisted deep eutectic solvents (DESs-UAE). The astaxanthin content was determined by high-performance liquid chromatography (HPLC). Six types of deep eutectic solvents composed of DL-menthol and selected hydrogen bond donors were prepared and evaluated, among which the DL-menthol–acetic acid system showed superior extraction performance. Response surface methodology (RSM) was employed to optimize extraction parameters (ultrasonic power, time, and temperature), and the optimal conditions were determined as follows: ultrasonic power 420 W, ultrasonic time 20 min, and ultrasonic temperature 60 °C, achieving an AST extraction rate of 62% (2.49 mg/g). Compared with conventional organic solvent extraction, DESs exhibited a significantly higher AST extraction rate from PR, except for dimethyl sulfoxide (DMSO). Scanning electron microscopy (SEM) analysis demonstrated that DES-UAE treatment disrupted the cellular structure of PR, resulting in numerous surface pores; this facilitated the release of intracellular bioactive components and significantly improved AST extraction efficiency. The PR extract showed no significant cytotoxicity and could effectively promote L929 cell proliferation. It concentration-dependently increased superoxide dismutase (SOD) activity and decreased malondialdehyde (MDA) content in H₂O₂-induced oxidative stress L929 cells, thereby alleviating oxidative damage. Additionally, it concentration-dependently upregulated type I collagen expression in these cells, ameliorated the decline in collagen synthesis function, and exerted a protective effect against cellular oxidative damage. This study provides a green alternative to toxic solvents and offers important theoretical and chemical support for the extraction of natural products and the high-value utilization of Phaffia rhodozyma (PR). Deep eutectic solvents have emerged as promising green alternatives to hazardous organic solvents, yet hydrophobic DESs tailored for lipophilic astaxanthin extraction from Phaffia rhodozyma and the linkage between extraction performance and anti-aging bioactivity remain insufficiently explored. Here, an ultrasound-assisted hydrophobic deep eutectic solvent extraction strategy was constructed to acquire astaxanthin, aiming to overcome low efficiency and environmental risks of conventional organic extraction techniques. Six DL-menthol-based DESs were prepared and screened, and DL-menthol–acetic acid possessed the optimal extraction capacity. Key extraction parameters were optimized via response surface methodology, and the maximum astaxanthin extraction recovery reached 62% (2.49 mg/g) under 420 W ultrasonic power, 20 min treatment and 60 °C. This yield was markedly higher than that of most common organic solvents; though comparable extraction effect was obtained with DMSO, the adopted DES possessed outstanding low-toxic and biodegradable superiorities that DMSO cannot match. SEM characterization verified that the combined treatment destroyed yeast cell structure and formed porous morphology, which accelerated intracellular astaxanthin release and accounted for improved extraction efficiency. Biological assays proved the extract possessed good biosafety and proliferation-promoting effect on L929 cells. It effectively relieved cellular oxidative injury by elevating the SOD level and reducing MDA accumulation in oxidative damaged cells, and upregulated type I collagen expression to mitigate aging-related collagen loss. This work develops an eco-friendly and high-efficiency extraction route for lipophilic active substance, confirms the practical value of hydrophobic DES, and provides experimental basis for high-value utilization of Phaffia rhodozyma resources. Full article

Essential oils (EOs) are widely investigated as multifunctional agents for cosmetic applications due to antimicrobial and antioxidant properties; however, their efficacy and safety depend on chemical composition and microbiological selectivity. This study characterized ten commercially available EOs (basil, lavender, melissa, mint, oregano, rosemary, sage, thyme, clove, tea tree) and evaluated their antistaphylococcal and antioxidant activities. Chemical composition was analyzed using GC-MS and ATR-FTIR spectroscopy, while antistaphylococcal activity was assessed against Staphylococcus spp. isolated from skin microbiota using the microbroth dilution method. Antioxidant capacity was determined by the DPPH assay. Multivariate statistical analyses were applied to explore chemical composition–activity relationships. The EOs showed distinct chemical profiles dominated by phenolic compounds (eugenol, carvacrol, thymol), oxygenated monoterpenes (linalool, menthol, 1,8-cineole), and terpenoids. Clove, oregano, and thyme EOs exhibited the strongest antistaphylococcal activity (MIC 0.5–2 mg/mL) and highest antioxidant capacity (>80%), whereas lavender, mint, and sage EOs showed weak effects. All EOs demonstrated bactericidal activity with consistent susceptibility patterns among Staphylococcus spp. PCA and FTIR confirmed an association between phenolic content and bioactivity. In conclusion, antimicrobial efficacy is primarily driven by phenolic composition, supporting targeted cosmetic use while acknowledging possible effects on skin Staphylococcus spp. populations. Full article

This study explores the use of microwave-assisted extraction of bioactive compounds from peppermint (Mentha piperita L.) using aqueous ethanolic and β-cyclodextrin solvents. The experiments were performed according to Response Surface Methodology, and the central composite design was employed, with the independent variables being treatment time, microwave power and solvent concentration. During the research, the total polyphenol content (TPC, expressed as gallic acid equivalents, GAE), total flavonoid content (TFC, expressed as quercetin equivalents, QUE), antioxidant capacity (AC) using the FRAP (Ferric Reducing Antioxidant Power, expressed as ascorbic acid equivalents, ASE) and DPPH (2,2-diphenyl-1-picrylhydrazyl) methods, and total menthol content (TMC, expressed as menthol equivalents, MEE) were examined. For the ethanolic extraction, the optimum conditions were 571 W, 7.3 min, and 10% v/v. The following results were obtained for these parameters: TPC, 1025 µg GAE/mL; TFC, 66.69 µg QUE/mL; FRAP, 3786.24 µg ASE/mL; DPPH, 80.24%; and TMC, 62.63 µg MEE/mL. In the case of β-cyclodextrin, the optimum conditions are 800 W, 11.81 min, and 1.70 mM. In this case, the results are as follows: TPC, 949.29 µg GAE/mL; TFC, 90.75 µg QUE/mL; FRAP, 4275.54 µg ASE/mL; DPPH, 89.81%; and TMC, 84.99 µg MEE/mL. β-cyclodextrin increased the flavonoid, antioxidant and menthol content, although there were no significant differences between the concentrations. The use of ethanol had a greater effect on the polyphenol content. β-cyclodextrin proved to be an effective green alternative solvent for peppermint. Full article

This study investigates the volatile composition of twelve medicinal plant species belonging to the Lamiaceae family, which are widely recognized for their diverse biological activities, including antioxidant, antibacterial, and antifungal properties. Despite extensive studies on essential oils, comparative analyses using solvent-free techniques under different microclimatic conditions remain limited. This study investigates the volatile compounds in twelve medicinal plants from the Lamiaceae family using headspace solid-phase microextraction coupled with gas chromatography–mass spectrometry (HS-SPME/GC-MS). Lamiaceae plants are recognized for their diverse medicinal properties, including antioxidative, antibacterial, and antifungal effects. A total of 74 volatile compounds were identified, encompassing terpenes, alcohols, esters, aldehydes, and ketones. Notably, Lavandula spica L. exhibited the highest number of unique volatiles (28), while Melissa officinalis L. had the fewest (16). Key compounds included Citral (65.48%) in Melissa officinalis L., Menthol (33.37%) and Menthyl acetate (30.53%) in Mentha piperita L., Carvone (45.86%) in Mentha spicata L., and Eucalyptol (54.71%) in Origanum syriacum L. Plants from Adana Botanic Park were rich in terpenes and ketones, whereas those from Osmaniye contained higher levels of alcohols, aldehydes, and esters. The findings emphasize the impact of geographic location on volatile profiles and suggest avenues for further research into medicinal efficacy and optimal dosage. This study supports the sustainable use of plant biodiversity (SDG 15) and highlights the importance of bioactive compounds for human health and well-being (SDG 3). Full article

This paper describes a simple and sensitive method for determining the content of five bisphenols at the µg L⁻¹ level in tea infusion samples. The procedure uses a polypropylene hollow fiber filled with a deep eutectic solvent as the acceptor phase and 1-octanol as the supported liquid membrane, and the optimal conditions for the three-phase stir bar microextraction method were established as follows: a menthol–octanoic acid mixture (1:1 molar ratio) as the hollow-fiber filling, an extraction time of 1 h, and 80 µL of methanol for analyte desorption. The method demonstrated good linearity over the ranges of 1.5–30.0 µg L⁻¹ (BPF, BPA, BPAF, and BADGE) and 6.0–120.0 µg L⁻¹ (BPZ), with limits of detection between 0.28 and 1.01 µg L⁻¹, and the relative recovery values were satisfactory (99–120%) with acceptable precision (RSD < 17%). Thus, the method was successfully applied to quantitatively analyze twenty commercial tea infusions, in which BPF was detected at concentrations above the LOQ, and the greenness and overall applicability of the approach were confirmed using the AGREEprep and BAGI assessment tools. Full article

Three-dimensional (3D) printing has emerged as a versatile platform in pharmaceutical sciences, enabling fabrication of personalized dosage forms with controlled drug release and tailored properties using printable hydrogel bioinks. This study aimed to develop mucoadhesive hydrogel buccal films for cannabinoid delivery using extrusion-based 3D bioprinting. The films incorporated cannabidiol (CBD) and tetrahydrocannabinol (THC) as cyclodextrin inclusion complexes with HPMC or Carbopol as mucoadhesive hydrogel-forming polymers, while terpenes were evaluated as permeation enhancers. Terpenes including 1,8-cineole, d-limonene, α-pinene, and L-menthol were investigated individually and in combinations to assess their ability to enhance buccal cannabinoid permeation. Hydrogel bioinks were prepared and characterized for viscosity, pH, and drug content prior to printing under optimized conditions. The printed films were evaluated for mechanical properties, swelling behaviour, mucoadhesion, in vitro drug release, and ex vivo buccal mucosal penetration. Ex vivo penetration studies demonstrated that combinations of natural terpenes significantly improved CBD penetration compared with individual terpenes and the synthetic enhancer Azone. HPMC-based hydrogel films exhibited superior mechanical strength, cohesive gel matrices, and sustained non-Fickian cannabinoid release, while enhancing transmucosal penetration compared with unformulated drugs. Carbopol-based films showed higher mucoadhesion but weaker mechanical properties and faster erosion-driven release. These findings demonstrate the potential of 3D-printed mucoadhesive hydrogel films as gel-based systems for transmucosal cannabinoid delivery. Full article

This study evaluated the use of HDES for omega-3 recovery from by-products of Octopus maya, an endemic species of the Yucatán Peninsula, Mexico. A 2 × 3 × 2 factorial design was applied to assess the effect of: (1) the hydrogen bond acceptor (HBA, menthol or eucalyptol) of hydrophobic deep eutectic solvents (HDES) with oleic acid as the hydrogen bond donor; (2) the molar ratio (MR) (1:1, 1:2, or 2:1); and (3) ultrasound-assisted extraction time (ET) (30 or 60 min) in omega-3 equivalents (EO3, mg/mL), determined by UV–Vis spectrophotometry and viscosity characterization to the Octopus maya extracted samples (n = 2), reported as $\overline{x}$ ± SD. The effects of the factors studied were analyzed by a DOE methodology with Minitab^® (version 18). Samples with the highest omega-3 were selected and their composition was confirmed by FTIR, Raman spectroscopy and gas chromatography. Eucalyptol at a molar ratio of 1:2 and an extraction time of 30 min yielded the highest OE3 (0.70 mg/mL). The statistical analysis revealed that the extraction of omega-3 determined by UV–Vis spectrophotometry was significantly influenced by the triple interaction of HBA × MR × ET (p < 0.05), indicating that extraction performance depends on the combined effect of solvent composition and processing conditions. All extracts showed Newtonian behavior with viscosities between 0.011 and 0.036 Pa·s, with eucalyptol formulations presenting the lowest values (0.011–0.023 Pa·s). Fatty acid profile allowed to quantify C16:0; C18:0; C18:1 n-9; C18:2 n-6; and C18:3 n-3, palmitic, stearic, oleic, linoleic and linolenic fatty acids, respectively, showing greater affinity for polyunsaturated fatty acids, mainly omega-6 (23.45–27.91%), and lower affinity for saturated fatty acids such as palmitic and stearic acids, indicating HDES as a sustainable alternative for selective extractions. Full article

In this study, new types of eutectic solvents (ESs) are tested for their ability to remove the eight most common bisphenols (BPA, BPB, BPC, BPE, BPF, BPG, BPS, BPAP), which are environmentally monitored substances, from aqueous matrices. A total of 18 ESs based on hydrophobic organic acids, such as capric, caprylic, lauric, and myristic acids, and terpenes, such as DL-menthol, terpineol, linalool, and geraniol, are prepared and mixed in various molar ratios. The highest extraction yield for all types of BPs is achieved with a binary mixture of geraniol:caprylic acid prepared in a molar ratio of 1:1. This ES can be used repeatedly for five consecutive cycles achieving almost 100% recovery for BPB, BPC, BPG, and BPAP, while for BPA, BPE, and BPF, the yield drops to 97% and for BPS to 90%. The efficiency of ES extraction is verified using HPLC-MS/MS to determine the BPs in the aqueous phase. This is performed at a pentafluorophenylpropyl stationary phase with LOQs ranging from 0.24 to 29.1 ng/mL. The applicability of this HPLC-MS/MS method was demonstrated by monitoring the occurrence of BPs in thermal paper and other industrial samples. Full article

Fusarium oxysporum f. sp. lycopersici (FOL) is one of the most destructive soil-borne pathogens affecting tomato production worldwide, causing substantial yield losses and persisting in soil for extended periods. The increasing regulatory restrictions on chemical fungicides and the emergence of resistant pathogen strains have intensified the search for sustainable and environmentally friendly alternatives. This systematic review synthesizes studies published between 2000 and 2025 that evaluated the antifungal efficacy of essential oils (EOs), their bioactive constituents, and EO-based nanoformulations against FOL in tomato. A total of 40 studies were included, following the PRISMA 2020 guidelines, encompassing in vitro, greenhouse, and limited field evaluations. Many EOs rich in phenolic compounds and oxygenated monoterpenes, such as thymol, carvacrol, eugenol, citral, and menthol, consistently inhibited FOL growth and spore germination, with reported mycelial growth inhibition ranging from 60 to 100% and minimum inhibitory concentrations (MICs) between 0.05 and 1.5 µL ml⁻¹. However, the use of EOs is often limited because they evaporate quickly, do not mix well with water, can harm plants, and do not persist under field conditions. Nano-delivery systems, including nanoemulsions, polymeric nanoparticles, chitosan-based carriers, and lipid-based nanostructures, have been shown to enhance the stability, bioavailability, and antifungal efficacy of EOs. This has led to improved disease management and reduced pesticide application rates. In addition, several EO-based treatments have been reported to activate plant defense responses, including the induction of defense-related genes, antioxidant enzymes, and epigenetic modifications. Overall, EO-based nanoformulations show promise as next-generation biopesticides for the sustainable management of tomato Fusarium wilt. Nevertheless, large-scale field validation, standardized formulation protocols, and regulatory assessments are required before these technologies can be widely implemented in agriculture. Full article

Objective: This study investigated the effects of menthol (MEN) mouth rinsing (MR) on cycling performance, neuromuscular activation, and perceptual responses during high-intensity exercise in the heat. Methods: A total of 10 trained adolescent male cyclists (16.7 ± 1.3 yrs; VO_2peak: 62.3 ± 7.6 mL·kg⁻¹·min⁻¹) completed a familiarization and two randomized, single-blind trials using a modified variable cycling test (M-VCT) in the heat (31.45 ± 0.59 °C; 23.40 ± 2.55% RH). The participants rinsed with 0.01% L-menthol or a placebo every 6 min during exercise. Power output (PO), cadence (RPM), rating of fatigue (ROF), affective feeling (FS), and surface electromyography (sEMG) were recorded. Results: Menthol MR significantly increased mean PO by 1.67 ± 1.59% (MEN: 177.1 ± 33.0 W; PLA: 174.1 ± 32.1 W; p = 0.002; d = 1.42) and enhanced cadence (MEN: 87.4 ± 5.1 RPM; PLA: 84.5 ± 5.2 RPM; p = 0.027; d = 0.84), particularly during high-intensity intervals. No significant differences were observed in ROF or FS between conditions (p > 0.05). Five muscles were monitored for activation (RF, VM, VL, TA, Gast). A significant main effect of time demonstrated decreased activation in VM, TA, RF, and Gast. VL showed a trend toward a main effect of condition (p = 0.057), with lower activation in MEN. Both VL and RF exhibited significant condition × lap interactions (p = 0.007 and p = 0.017), with progressively lower activation in MEN as fatigue progressed. Conclusions: Menthol MR significantly improved cycling performance in the heat without altering perceptual or physiological strain. Some muscles demonstrated reduced activation with menthol MR, and further study is needed to confirm the magnitude of ergogenic effects and elucidate the physiological mechanism. Full article

Cannabis sativa L. leaves (CSL) are a rich in bioactive compounds and known for their medicinal and recreational uses. In this study, a natural hydrophobic deep eutectic solvent (HDES) system composed of menthol and thymol (1:1) was employed for the efficient extraction of bioactive compounds from CSL. Extraction of bioactives was optimized at various conditions involving DES/ethanol ratio, temperature, and extraction time, as well as shaking speed through statistical models including response surface methodology (RSM) and artificial neural network (ANN). The maximum bioactive yield, equal to 70% (w/w) of powdered CSL, was achieved at optimized values of 5.5 mL DES, 4.5 mL ethanol, and 225 rpm shaking speed at 55 °C for 107.5 min. It was observed that slightly adjusting the shaking speed and temperatures customized the nature of bioactives with more antioxidant, antidiabetic, and antimicrobial properties. The extracts of CSL produced while applying natural HDES were found to be non-toxic during hemolytic assay. Overall, HDES when mixed with ethanol in 55:45 ratio produced CSL extracts with an ample level of phenolics (133.75 mg GAE/g) and flavonoids (120.05 mg QE/g). GC-MS analysis of CSL extracts produced by HDES revealed the presence of multiple bioactives like tetrahydrocannabivarin, cannabidiol, cannabinol, cannabidivarol, dl-menthol, levomenthol, and 4-hydroxy-3-methylacetophenone. Based on these findings, it can be concluded that HDES in combination with ethanol may work as an efficient extraction solvent to recover CSL bioactives without compromising their antioxidant features and safety for use in food and pharmaceutical applications. Full article

Peppermint tea is one of the most widely consumed herbal beverages worldwide, yet limited data are available on the chemical variability and quality of commercial products. This study assessed the essential oil (EO) yield and composition of 39 commercial peppermint (Mentha × piperita L.) teas and commercial EOs obtained from different countries. EO yield showed pronounced variability, ranging from 0.8 to 26.8 mL/kg. In total, 112 compounds were identified by GC-MS, accounting for 88.4–99.5% of the total EO composition. The major constituents were menthol (0.1–25.2%), menthone (0.1–21.8%), and carvone (0.6–30.0%), revealing marked chemical heterogeneity among marketed products. Several samples exhibited unusually high carvone levels accompanied by reduced menthol and menthone contents, reflecting substantial chemotypic diversity and inconsistency among marketed products. The concentrations of the regulated constituents, pulegone (0.02–2.56%) and menthofuran (0.02–6.58%), remained within the European Pharmacopoeia limits. Based on the observed levels of pulegone and menthofuran, the results do not indicate a safety concern for consumers under normal tea preparation and consumption conditions. Overall, the findings demonstrate substantial variability in the quality of commercial peppermint teas and highlight the relevance of essential oil profiling as a tool for assessing the quality and safety of herbal beverages. Full article

This paper reports the chiral separation of menthol enantiomers using the VARICOL process to improve productivity. Amylose 3,5-dimethylphenylcarbamate coated on silica gel was employed as the chiral stationary phase, and n-hexane/2-propanol (95/5, v/v) was used as the eluent. To design and optimize the VARICOL process, a linear driving-force model was developed to predict the separation performance. Separation regions of the conventional simulated moving bed (SMB) and VARICOL processes were evaluated and compared. It was found that, under an outlet purity requirement of 95.0%, the five-column VARICOL process has a separation region comparable to that of the six-column conventional SMB process. As an illustrative example, a five-column VARICOL unit and a six-column conventional SMB unit, both operating under the same conditions, were employed to resolve the menthol racemate. Purities for both the extract and raffinate were above 95.0%, and a productivity of 0.400 g_racemate/(L_CSP∙min) and a solvent consumption of 0.355 L/g_racemate were achieved in the VARICOL process. Productivity increased by 20% while solvent consumption maintained relative to the conventional SMB process, though product purities decreased slightly. Full article