Search Results (283)

Oak (Quercus spp.), traditionally used for aging alcoholic beverages, is not native in many countries, which increases production costs and environmental impact. During the aging process of alcoholic beverages, complex physical and chemical transformations occur that determine their chemical composition and sensory quality, many of which are unique depending on the type of wood used in the process. In this context, Maclura tinctoria (Tajuva), a native Brazilian species rich in phenolic compounds, was evaluated based on its phenolic composition and extraction behavior as a sustainable alternative for beverage aging. Wood chips were subjected to three toasting levels (untoasted, medium, and high) and aged for up to 360 days in two hydroethanolic model systems (10% and 14% v/v ethanol). The total and individual phenolic compounds were determined using the Folin–Ciocalteu method and HPLC–DAD/LC–MS/MS analysis. Results showed that toasting level, ethanol concentration, and aging time significantly influenced phenolic extraction. Untoasted Tajuva released the highest amounts of phenolic acids and flavonoids, particularly gallic and caffeic acids, and quercetin, respectively; while medium toasting favored the formation of thermally derived aromatic compounds, such as vanillic acid. The 14% ethanol system enhanced extraction efficiency for most analytes. Overall, Tajuva wood exhibited higher phenolic yields than French oak under comparable conditions, highlighting its chemical richness and extraction reactivity. These findings support the use of M. tinctoria as an eco-efficient and functional alternative to oak for the maturation of alcoholic beverages. Full article

This multidisciplinary study investigates Piper rivinoides, a Brazilian medicinal species, focusing on its chemical composition and antimycobacterial potential. UHPLC-HRMS/MS of leaves, stems, branches, and roots revealed 58 compounds, including neolignans, lignanamides, triterpenes, flavonoids, and carotenoids. Fourteen metabolites, notably benzofuran neolignans and pentacyclic triterpenes are annotated here for the first time. Quantitative analyses by HPLC-DAD-UV showed that eupomatenoid-5, eupomatenoid-6, and conocarpan were most abundant in leaves, reaching amounts approximately twice those found in branches and stems and about ten times higher than in roots, supporting the optimal defense theory and organ-specific accumulation of bioactive metabolites. Biological assays against Mycobacterium tuberculosis strains H37Rv and M299 revealed strong inhibitory activity for the leaf extract and isolated neolignans. Eupomatenoid-5 and eupomatenoid-6 achieved inhibition comparable to rifampicin, with low MIC₅₀ values, while conocarpan exhibited moderate activity. Antimycobacterial effects were more pronounced against the H37Rv strain, although relevant inhibition was also observed for the hypervirulent M299 strain. These findings highlight P. rivinoides as a rich source of benzofuran neolignans and promising antimycobacterial properties. The integration of advanced mass spectrometric analyses with bioassays demonstrates the value of combining chemical and biological approaches to uncover novel natural products. The putative identification of new neolignans and triterpenes, along with the confirmation of potent antimycobacterial activity, provides a robust foundation for further studies on biosynthesis, structure–activity relationships, and potential biotechnological applications. Full article

Skin sensitization is a critical endpoint in cosmetic safety assessment, necessitating reliable animal-free testing alternatives. Current established in chemico assays, such as the Direct Peptide Reactivity Assay and Amino acid Derivative Reactivity Assay, are limited by prolonged 24 h incubation periods and their inability to distinguish between direct electrophilic sensitizers and pro-electrophiles requiring metabolic activation or spontaneous oxidation. This study presents the design, synthesis, and validation of NNDNAC (N,N-dimethyl N-(2-(1-naphthyl)acetyl)-l-cysteine), a novel nucleophilic reactivity probe synthesized via a seven-step pathway. A modified naphthalene structure featuring N,N-dimethylamino substituent enhances nucleophilicity of the cysteine sulfur atom, enabling rapid reactivity assessment within an hour incubation using LC-DAD quantification. Comparative validation studies demonstrated that NNDNAC rapidly identified strong electrophilic sensitizers, achieving 100% and 98% depletion rates for p-benzoquinone and 2-methyl-4-isothiazolin-3-one, respectively, within 1 h. Critically, the NNDNAC assay successfully differentiated pro-electrophiles like p-phenylenediamine and 4-aminophenol, which showed negligible depletion at 1 h but significant depletion after 24 h due to auto-oxidation. Furthermore, NNDNAC classified farnesal as a weak sensitizer, aligning with established KeratinoSens™ and LLNA data. The NNDNAC probe represents a significant advancement in skin sensitization assessment, offering a time-efficient, high-throughput platform that not only accelerates screening processes but also provides crucial mechanistic insights through electrophile/pro-electrophile differentiation, significantly improving animal-free toxicological evaluations. Full article

This study provides the first comprehensive chemical and biological profiling of Citrus lumia Risso & Poit. var. ‘Pyriformis’, a rare Mediterranean Citrus variety with unexplored nutraceutical potential. The fresh juice (CLPJ) showed a distinctive phytochemical composition, with 38.8 ± 0.99 mg gallic acid equivalents/100 mL of total phenols and 25.96 ± 2.37 mg rutin equivalents/100 mL of flavonoids. High-performance liquid chromatography coupled with diode-array detection (HPLC-DAD) quantification revealed high levels of organic acids, including ascorbic acid (0.34 g/L) and citric acid (34.6 g/L). Liquid chromatography coupled with diode-array detection and electrospray ionization tandem mass spectrometry (LC-DAD-ESI-MS/MS) enabled the annotation of 28 polyphenolic constituents, featuring glycosylated flavanones and several uncommon flavonols and acylglycosidic derivatives whose structural patterns are typical of primitive Citrus lineages and largely absent in commercial cultivars. Functionally, CLPJ exhibited multi-target antioxidant and anti-inflammatory activities and promoted epithelial repair in Caco-2 cells without cytotoxic effects. Overall, the juice displays a distinctive chemotaxonomic fingerprint and promising multifunctional properties, supporting its potential as a functional food ingredient and contributing to the valorization of minor Mediterranean Citrus biodiversity. Full article

Plant materials rich in proanthocyanidins are fractionated to determine the structure of these compounds and relate it to bioactivity. The aim of this study was to fractionate a procyanidin-rich hawthorn bark extract using low-pressure liquid chromatography and to determine the compound profile and antioxidant activity of the obtained fractions. We identified and quantified the phenolics of four fractions (I–IV) separated on a Toyopearl HW-40S column with methanol as the mobile phase, using HPLC-DAD and LC-ESI-MS techniques. The antioxidant activity was determined to comprise ABTS^•+ and DPPH^• scavenging activity, ferric-reducing antioxidant power (FRAP), and inhibition of β-carotene-linoleic acid emulsion oxidation. Characteristic data were subjected to principal component analysis (PCA). Fraction I contained mainly (−)-epicatechin (741.3 mg/g) and a lower amount of flavones and quercetin derivatives (100.7 mg/g). Fraction II was almost pure procyanidin B2, which accounted for 88.8% of the total phenolics. The subsequent fractions were rich in B-type procyanidin dimers, trimers, and tetramers. FRAP and antiradical activity against ABTS^•+ and DPPH^• of the fraction containing low-molecular weight phenolics was lower than those of the fractions with procyanidin oligomers. The antioxidant activity of fractions II–IV ranged from 8.95 to 9.28 and from 6.45 to 6.71 mmol TE/g in the ABTS and DPPH assays, respectively. Their FRAP was in the range of 17.67–21.06 mmol Fe²⁺/g. According to PCA, the procyanidin dimers of fractions II and III were associated with antioxidant activity in these assays. In turn, the procyanidins with the highest degree of polymerization (trimers and tetramers) present in fraction IV were related to the antioxidant activity measured in the β-carotene-linoleic acid emulsion system. Overall, the separation of purified hawthorn bark extract using low-pressure Toyopearl HW-40S column chromatography resulted in a fraction rich in procyanidin B2, as well as fractions containing procyanidins with an increasing degree of polymerization, all with high levels of antioxidant activity under various conditions and the potential for future applications in food, pharmaceuticals, and cosmetics products. Full article

(1) Background: Caffeine is one of the most widely consumed psychoactive substances. Its safety profile and short half-life make it an ideal drug model for studying the pharmacokinetics of caffeine. This study aimed to develop a method for determination of caffeine in a small volume of saliva (200 µL). (2) Methods: Solid-phase extraction was employed to isolate caffeine from saliva, followed by quantitative analysis using liquid chromatography coupled with diode-array detection. Chromatographic separation was achieved on a C18 column, using a gradient mobile phase of acetonitrile and 0.1% formic acid. (3) Results: The method was validated for selectivity, linearity, precision, and accuracy. Linearity was established over the range of 10–10,000 ng/mL (R² = 0.995). The coefficients of variation for intra- and inter-day precision for the three tested caffeine concentrations did not exceed 12.11%. Recovery from spiked saliva samples exceeded 90.53%. The developed method was applied to preliminary studies to follow the pharmacokinetics of caffeine in saliva. The concentration of the substance was studied in the saliva obtained from a volunteer after espresso consumption. (4) Conclusions: The developed method will offer a reliable approach for non-invasive caffeine monitoring in clinical and research applications. Full article

Merlot red wines rank among the most distinguished varietals globally. This study aimed to characterize the phenolic compound profiles of Merlot wines and assess the influence of geographical origin and vintage on samples from two Albanian wine regions. Using liquid chromatography coupled with tandem mass spectrometry, a total of 31 phenolic compounds were identified and quantified. These were classified into hydroxybenzoic acids and flavan-3-ols (13 compounds), phenolic acids (9), flavonols (5), and stilbenoids (4). The total phenolic content ranged from 294 mg L⁻¹ in wines from the Mati–Mirdita region to 480 mg L⁻¹ in those from the Durrës–Kavaja region, demonstrating significant regional variation. Notably, the hydroxybenzoic acids and flavan-3-ols exhibited the most pronounced differences, with gallic acid concentrations varying from 123 mg L⁻¹ (Mati–Mirdita) to 170 mg L⁻¹ (Durrës–Kavaja). Both regions’ wines were rich in catechin, epicatechin, procyanidin derivatives, trans-caftaric acid, and ethyl gallate. However, procyanidins were found in higher concentrations in the Mati–Mirdita wines, while other phenolics were more abundant in Durrës–Kavaja samples. These findings underscore the influence of geographical and climatic factors on phenolic composition, offering a robust chemical fingerprinting approach for assessing wine authenticity and quality. Full article

Rhododendron tomentosum Harmaja is a marsh plant known for its high content of bioactive components, including essential oil, flavonoids, and phenolic acids. In the current work, the effects of cultivation mode (agar, liquid stationary, shake flask, and temporary immersion) and experiment duration (30, 60, and 90 days) on the growth and contents of non-volatile phenolics in Rhododendron tomentosum microshoots were investigated. Agar and liquid stationary cultures provided the highest dry biomass yield per liter, but their dry weight productivities per day were the lowest among the tested systems. Agitated and temporary immersion cultures, on the other hand, were the most productive in terms of fresh and dry biomass yield per day. LC-DAD-ESI-MS analysis of extracts from microshoots and wild-grown plants revealed similarities in phenolic composition: in both cases, the presence of catechin, chlorogenic acid, and flavonoids of flavonol type (mainly glycosides of quercetin and myricetin) was confirmed. The qualitative composition of the phenolic fraction was not affected by experiment duration and cultivation mode. As determined by HPLC analysis, shake flask and temporary immersion cultures were characterized by the highest phenolic contents: up to 37.5 and 26 mg/g dry weight, respectively. The maximum productivities of the above systems were equal to 18 and 13.5 mg/L/d, respectively. Full article

Background: Crataegus almaatensis, an endemic hawthorn species from Kazakhstan, is known for its rich content of phenolic compounds and flavonoids with significant pharmacological potential. This study aimed to optimize and compare conventional solid–liquid extraction (SLE) and ultrasound-assisted extraction (UAE) processes for maximizing the extractability of bioactive compounds from hawthorn leaves powder. Methods: The effects of temperature, extraction time, ethanol concentration, and solid-to-liquid ratio (or ultrasound power in the case of UAE) on total phenolic content (TPC), total flavonoid content (TFC), and antioxidant activity (FRAP, DPPH, and ABTS assays) were systematically evaluated. Results: The UAE method yielded higher concentrations of TPC and TFC, with up to 16% improvement in TPC and reduced ethanol usage (40% (v/v)) compared to SLE (75% (v/v)), demonstrating its efficiency and sustainability. Optimal extraction conditions were identified as 70 °C, 75% ethanol, 34 min, and an S/L ratio of 0.05 g/mL for SLE, 70 °C, 40% ethanol, 44 min, and 100 W US power for UAE. High-resolution HPLC-DAD and LC-Q/TOF-MS analyses confirmed the presence of key phenolic acids and flavonoid glycosides, including chlorogenic acid and apigenin-8-C-glucoside-2′-rhamnoside as the most abundant compounds identified. Conclusions: These findings validate UAE as an innovative, eco-friendly method for extracting bioactive compounds from hawthorn leaves and highlight its potential for developing natural antioxidants for pharmaceutical and nutraceutical applications. Full article

(1) Background: The concentration of lamotrigine, an antiepileptic drug very often used in bipolar disorder, is most often determined in the blood, with many inconveniences. An alternative may be to use saliva as a diagnostic material for this purpose. The development of a method to determine lamotrigine in saliva as a biological material significantly improves patient comfort during sampling. The developed method uses solid-phase extraction for the isolation of the drug from saliva for the first time. (2) Methods: This study aimed to develop a method to determine lamotrigine in saliva using solid-phase extraction (SPE) for isolation and liquid chromatography with a diode array detector (LC-DAD) for quantitative analysis. (3) Results: The method was validated by determining its linearity in the concentration range 10–2000 ng/mL (R² > 0.99), and the intra- and inter-day precision expressed as coefficient of variation (CV%) did not exceed 15%. (4) Conclusions: The developed method was used to determine the salivary concentration of lamotrigine in patients treated with the studied compound, confirming its usefulness in bipolar disorder (BD). Full article

Colorectal cancer (CRC) is the third most diagnosed cancer worldwide, and p53 dysfunction plays a significant role in its pathogenesis by impairing cell cycle control and apoptosis. This study aimed to elucidate the phytochemical composition and anticancer potential of extract of residue from clove hydrodistillation (Syzygium aromaticum, SA) and seed extract from Syzygium nervosum (SN). LC-DAD-MS/MS analysis identified gallic acid (2.68%) and ellagic acid (6.70%) as major constituents in SA, while SN contained gallic acid (0.26%), ellagic acid (3.06%), and 2′,4′-dihydroxy-6′-methoxy-3′,5′-dimethylchalcone (DMC) as major constituents. Both extracts exhibited potent antioxidant effects as evidenced by DPPH and ABTS assays. In vitro assays showed that SA and SN significantly inhibited the proliferation of HCT116 (p53 wild-type) colorectal cancer cells, with minimal effects on HT-29 (p53 mutant) cells. Apoptosis was confirmed in HCT116 via Annexin V-FITC/PI staining and increased caspase-3/7 activity. Cell cycle analysis revealed sub-G1 accumulation, accompanied by upregulated p21 and concurrently downregulated cyclin D1 expression, both hallmarks of p53-mediated checkpoint activation. These molecular effects were not observed in HT-29 cells. In conclusion, SA and SN extracts selectively induce apoptosis and cell cycle arrest in p53-functional CRC cells, likely mediated by their phenolic constituents. These findings support their potential as promising plant-derived therapeutic agents for targeted colorectal cancer treatment. Full article

Among the craft recipes for artisans collected in the 4th-century Egyptian documents the Leyden and Stockholm papyri, there is one calling for adding animal milk to orchil for wool dyeing. To understand the rationale for this practice, wool yarns were dyed with and without goat milk added to orchil dyebaths, each made using lichens from three different sources. The results showed orchil containing milk dyed yarns a noticeably deeper red hue. The colorants extracted from the dyed yarns were analyzed by liquid chromatography-diode-array-detector-mass spectrometry to assess the relative amounts of nine identifiable orceins. The data showed that the yarns dyed with milk gave extracts exhibiting several fold more α-aminoorcein and α-hydroxyorcein, with only small differences in the other seven. Scanning electron microscopic analysis of a representative pair of dyed yarns showed that milk promoted surface changes in the fiber that may indicate increased cutaneous damage. Hypotheses for the milk’s effects on orchil dyeing were proposed that included the formation of milk–protein complexes with the two enriched orceins that possibly enhanced wool binding and/or better wool uptake of free and/or complexed orceins due to biodegradation of the wool’s surface cuticle caused by microbial growth promoted by the addition of milk. Full article

To address the need for improved nutritional value of organically grown wheat, this study investigated the impact of silicon treatments (AdeSil, ZumSil) on yield, health status, and bioactive compound content in spring wheat cultivars. The 2019–2020 research evaluated different application variants: seed dressing, foliar sprays, and their combinations. Comprehensive seed dressing combined with two foliar treatments, (variant B) and two foliar treatments (variant C), significantly increased yield (by an average of 8.9% and 7.6% vs. control, respectively). These variants beneficially affected fungal disease resistance mainly in the stressful 2019; in optimal 2020, they showed no clear advantage over the control, which performed similarly or better. Seed dressing (variant D) increased total phenolic acids (PAs) content and antioxidant activity, with the spelt cv. Wirtas exhibiting the highest levels. Silicon treatments modified alkylresorcinols (ARs) content, but effects depended on the year, cultivar, and application variant, not always exceeding the control. Silicon treatments, especially when applied in combination (seed dressing and foliar application), can improve spring wheat yield and favorably modify PAs content, enhancing grain nutritional value. However, the plant response regarding health status and ARs content is strongly conditioned by cultivar specificity and the prevailing environmental conditions of the growing year. Full article

Ethnographic sources provide information about several dye plants that produced reddish colors; however, there is no information on how this process is accomplished. Combining information from written sources with the results of dyeing experiments enables a deeper understanding of the dyeing methods employed in the past. This paper gives insight into the effect of using alkali on obtaining reddish tones in dyeing with Potentilla erecta and Rumex sp. In dyeing experiments, wool yarn was dyed with plant extracts, and the chemical compositions were studied both in plant extracts and in extracts obtained from wool after dyeing. As a result, the red/red-brown color is obtained only under the influence of alkali. Analytical studies of procyanidin (PC) extracts from rhizomes and yarns were performed using infrared spectroscopy (FTIR-ATR) and liquid chromatography coupled with mass spectrometric detection (LC-DAD-MS). Procyanidin extracts of P. erecta and R. acetosa contained monomers identified as (+)-catechin and (-)-epicatechin, as well as dimeric procyanidins of type-A (m/z 575 [M−H]⁻) and type-B (m/z 577 [M−H]⁻), along with various types of trimers (m/z 865 [M−H]⁻; m/z 863 [M−H]⁻), which were also isolated from dyed wool yarns with a similar composition. The conducted research on the use of alkali with tannin-containing plants contributes to deepening our understanding of the perception of color that existed in the ancient rural environment. Full article

We report the development of a procedure for ultrasound-assisted microscale extraction of metabolites from the flowers of Saint John’s wort (Hypericum perforatum L.), designed for comparative metabolite analysis of plants from genetic resource collections and natural and segregating populations. The procedure involves high-throughput methanol extraction of metabolites from ground-frozen flowers at a selected stage of flower development, which is carried out in a standard 2 mL Eppendorf tube. A total of 18 compounds, including chlorogenic acid, catechins, glycosylated flavonoids, hypericins, and hyperforin, were identified based on LC/DAD/QTOF analysis, of which 16 could be detected in the UV-Vis spectrum. Two alternative versions of the procedure were evaluated: the “single-flower” procedure, including repeated collection and analysis of single flowers from the tested plant, and the “bulk-flower” procedure, employing the collection of a bulk flower sample from the tested plant and analysis of a portion of the ground sample. The results showed excellent technical reproducibility of the “single-flower” procedure when used with the suggested combination of the peak areas for the proto- and stable forms of pseudohypericin and hypericin. Application of the developed “single-flower” procedure for comparison of the plants derived from seed progeny of the apomictic line Hp93 revealed significantly lower metabolite variation among the apomictic progeny plants compared to the variation observed among plants belonging to different genotypes. Full article