Search Results (1,353)

Black garlic peels (BGPs) are a largely underutilized by-product despite representing an untapped source of bioactive compounds. This study presents a sustainable upcycling protocol for black garlic peels, evaluating natural deep eutectic solvents (NaDES) to develop multifunctional extracts for green cosmetics. Following the screening of four eutectic mixtures, the choline chloride–lactic acid (ChCl:LA) system demonstrated the highest extraction efficiency. The optimized extract yielded a remarkable total phenolic content (5216.61 µg GAE/mL) and strong antioxidant capacity, confirmed by DPPH and FRAP assays, associated with recovering both free and bound phenolic fractions. Subsequent HPLC profiling characterized the extract, and the comparative analysis explicitly demonstrated that antimicrobial activity is entirely driven by and identical to the pure eutectic solvent vehicle rather than the extracted garlic biomass, with broad-spectrum efficacy against C. albicans, P. aeruginosa, and S. aureus. To evaluate its cosmeceutical potential, the extract was incorporated into emulsions (5%, 10%, 15% w/w) with inorganic or organic UV filters. Although the direct Sun Protection Factor (SPF) and the UVA Protection Factor (UVA-PF) did not show enhancing results, a photochemiluminescence (PCL) analysis revealed a synergistic behaviour with organic filters, successfully boosting the formulation’s biological antioxidant shield. This pioneering work highlights BGP’s upcycling potential, proposing NaDES extracts as highly promising multifunctional, antioxidant, and antimicrobial ingredients for next-generation cosmeceuticals. Full article

A normal-phase high-performance liquid chromatography (HPLC) method with UV spectrophotometric detector (SPD) detection was established for the simultaneous analysis of four tocopherol (TP) homologues and their corresponding tocopherol quinones (TQs). Separation was performed on a Sepax Technologies silica column (4.6 mm × 250 mm, 5 μm) using a mobile phase consisting of n-hexane, isopropanol, and tetrahydrofuran, allowing the eight target compounds to be effectively separated within 15 min. Method validation was conducted using standard solutions prepared in n-hexane, providing a direct evaluation of the chromatographic performance under a relatively clean solvent system. The method showed satisfactory linearity, with R² values of 0.9850–0.9996. The limit of detection (LOD) and limit of quantification (LOQ) ranges were 0.140–0.371 and 0.467–1.235 μg/mL for TP homologues and 0.0564–0.0856 and 0.1708–0.2595 μg/mL for TQ homologues, respectively. Precision was acceptable, with intra-day and inter-day relative standard deviations (RSD) below 2.69% and 3.78%, respectively. To further evaluate its applicability in oil matrices, recovery experiments were performed in peanut oil and camellia oil, yielding recoveries of 91.7–105.8% and 90.9–97.7% for TP homologues and 82.8–98.7% and 79.5–101.8% for TQ homologues, respectively. The method was further applied to edible oil samples. The results showed that the major TP homologues and detectable TQ homologues were determined, while some homologues were below the detection limits. Overall, the proposed method offers a simple and practical approach for the simultaneous analysis of TP and TQ homologues, thereby supporting further investigation of tocopherol oxidation in edible oils. Full article

Yellow onion (Allium cepa L.) outer skins are a high-volume agricultural waste that can be converted into commercially valuable bioproducts using various extraction techniques. This research focused on optimizing a green ultrasound-assisted extraction (UAE) method which allows for the isolation of several phytochemicals valued for their health benefits, such as polyphenols and flavonoids. HPLC/UV analysis of the extracts showed that the main component was quercetin. A one-factor-at-a-time (OFAT) design was used to identify the extraction parameters needed in order to maximize the amount of extracted target phytochemicals. The polyphenols, flavonoids and quercetin contents, along with the antioxidant activity of the extracts, were optimized by response surface methodology using a Box–Behnken design. Ultrasound amplitude, ethanol concentration, and time were selected as the most appropriate variables. The final results showed that TPC ranged from 78.16 to 97.16 mg GAE/g DM, TFC ranged from 22.77 to 26.46 mg QE/g DM, while CUPRAC values varied between 145.24 and 163.75 mg TE/g DM. The optimal extraction conditions were determined using a Box–Behnken model as 30% ultrasound amplitude, 53% ethanol concentration, and an extraction time of 13 min. The use of these conditions allowed the TPC, TFC and CUPRAC to show predicted values of 97.8 mg GAE/g DM, 27.2 mg QE/g DM, and 159.8 mg TE/g DM, respectively. These findings indicate that onion skin extracts could represent a green and promising source of antioxidant phytochemicals. Full article

Background: Predictive modeling of vaccine stability is an essential tool for accelerating development and ensuring product quality, particularly when long-term data are limited. To ensure high-quality input for accurate stability predictions, it is often necessary to allocate substantial analytical resources. Methods: This study demonstrates that integrating high-throughput screening (HTS) techniques such as ultraviolet–visible spectrophotometry (UV-VIS), intrinsic fluorescence and dynamic light scattering (DLS) with Advanced Kinetics Modeling (AKM) results in a synergistic approach, facilitating the development of robust predictive stability models. Results: Here, we applied AKM to a glycoconjugate vaccine targeting extra-intestinal pathogenic Escherichia coli (ExPEC). The aggregation processes observed via size-exclusion high-performance liquid chromatography (SE-HPLC), DLS, turbidity by UV-Vis absorbance and local changes in tryptophan microenvironment captured by intrinsic fluorescence were effectively described by the developed kinetic models. Conclusions: Using accelerated stability data across multiple temperatures, AKM successfully described key degradation pathways. Furthermore, the HTS assay results showed strong correlation with SE-HPLC data, indicating that these assays provide an efficient alternative requiring minimal analytical resources, material, and time. Full article

Background/Objectives: Coffea canephora (robusta) and Coffea liberica var. dewevrei (excelsa) cultivated in the Comoros islands represent understudied coffee varieties grown in a unique volcanic terroir. Despite their agricultural significance and potential bioactive value, no comprehensive biochemical or nutritional characterization of these Comorian coffees had previously been conducted. This study therefore aimed to provide the first integrated biochemical and nutritional characterization of both varieties and to evaluate the influence of the islands’ specific edaphoclimatic conditions on their chemical composition. Methods: An integrated analytical approach was employed, combining UV-Vis spectrophotometry, HPLC, ionomics, and FTIR-ATR spectroscopy to quantify polyphenols, flavonoids, caffeine, soluble sugars, antioxidant activity, mineral profiles, and macromolecular composition of green coffee beans from both species. Results: Robusta exhibited significantly higher levels of total polyphenols (121.79 ± 2.73 mg GAE/g), total flavonoids (29.43 ± 2.20 mg QE/g), caffeine (1.52% w/w), total soluble sugars (60.47 ± 3.37 mg GE/g), and antioxidant activity (64.97 ± 6.25 mM Trolox eq/g). Conversely, excelsa demonstrated a distinct mineral profile, with significantly higher concentrations of magnesium, calcium, sodium, zinc, and copper. FTIR spectroscopy confirmed distinct vibrational fingerprints between the two species, particularly in lipid and carbohydrate signatures. Conclusions: These findings position Comorian robusta as a potent source of antioxidants and stimulants, while excelsa offers a nutritionally balanced profile with nutraceutical potential, providing a scientific basis for valorizing both varieties as high-value niche products and contributing to the preservation of coffee agro-biodiversity. Full article

Mesenchymal stromal/stem cells (MSCs) are increasingly explored for immune-mediated diseases, yet standardized analytical readouts that capture coordinated immunomodulatory output across complementary secretory pathways remain limited. Here, we report the feasibility of an HPLC-based multi-analyte secretome characterization panel that quantifies two small-molecule outputs—adenosine and kynurenine—alongside two immunomodulatory proteins—interleukin-10 (IL-10) and transforming growth factor-beta (TGF-β)—in conditioned media from canine adipose-derived MSCs (cAD-MSCs). Canine immune-mediated hemolytic anemia (IMHA) was used as a disease context to motivate the selection of these analytes, given the pro-inflammatory cytokine environment characteristic of this condition. Three independent cAD-MSC lines were evaluated under baseline conditions and following cytokine stimulation with recombinant interferon-gamma (IFN-γ; 100 ng/mL) and tumor necrosis factor-alpha (TNF-α; 50 ng/mL), referred to herein as inflammatory priming or licensing. Conditioned media were collected at 72 h for metabolite analysis and 48 h for protein analysis, and quantified by HPLC using external calibration and peak integration. Across all three lines, licensing produced directionally consistent increases: mean adenosine increased 2.3-fold, mean kynurenine increased 3.1-fold, mean IL-10 increased 1.6-fold, and mean TGF-β increased 1.7-fold compared with unlicensed controls. Metabolite measurements for adenosine and kynurenine are reported with full chromatographic selectivity data; IL-10 and TGF-β measurements by reversed-phase HPLC with UV detection are presented as exploratory/semi-quantitative outputs and will require orthogonal confirmation (e.g., immunoassay) in future work. These findings are preliminary, derived from three independent donor lines with no comparator group, and are intended to support feasibility of the analytical framework rather than establish definitive performance specifications. Collectively, the data support the potential of a multi-analyte HPLC-based characterization panel to capture licensing-responsive secretory shifts across mechanistically complementary pathways, providing a foundation for expanded development and validation. Full article

Improving the dissolution and solubility of poorly water-soluble drugs remains a major challenge in drug development. Solid dispersion (SD) techniques offer an effective strategy by which to enhance the bioavailability of BCS Class II drugs such as ivermectin (IVM). This study aimed to develop and validate stability-indicating analytical methods for the quantification of IVM and to evaluate the performance of the formulated SDs. A novel RP-HPLC and a UV spectrophotometric method were developed and validated in accordance with ICH guidelines. IVM SDs were prepared via physical mixing (PM), solvent evaporation (SE), and melt fusion (MF) using Poloxamer 188, Kollicoat^® IR, and PEG 6000 at respective ratios of 1:1, 1:3, and 1:5. Dissolution studies showed a marked enhancement in drug release from SDs prepared by SE and MF methods compared with pure IVM. Among all formulations, the Poloxamer 188-based binary SD prepared by the SE method at a 1:5 ratio exhibited the highest dissolution (98.55% at 60 min), with release kinetics following anomalous (non-Fickian) transport (n = 0.681) according to the Korsmeyer–Peppas model. Solid-state characterization evidenced by FTIR, DSC, TGA, and SEM confirmed the transformation of IVM from its crystalline form to an amorphous state. Future studies will focus on the in vivo evaluation of the optimized IVM SD formulations. Full article

Ozone layer depletion exacerbates UV-induced skin damage, including oxidative stress and DNA lesions, thereby increasing the risk of photoaging and malignant transformation. Natural extracts have gained increasing attention as a photoprotective ingredient in cosmeceutical products. Kaempferia galanga, a species in the Zingiberaceae family traditionally used for skin-related treatment and listed in the CosIng database, exhibits multiple biologically relevant properties; however, its anti-photoaging and anti-photo-senescence effects in human dermal fibroblasts remain unexplored. This study investigated the in vitro photoprotective effects of K. galanga extract against UVB-induced photoaging and cellular senescence in human dermal fibroblasts. The ethanolic extract of K. galanga rhizomes (EKGRs) contained ethyl p-methoxycinnamate (EPMC) as a major constituent (33.7 ± 3.7% (w/w) of the crude extract), identified by HPLC-UV. Additionally, EKGR exhibited significant protective effects in UVB-irradiated fibroblasts. EKGR showed no cytotoxicity at concentrations up to 50.0 µg/mL, as determined by the MTT assay. EKGR pretreatment significantly reduced UVB-induced cellular senescence in human dermal fibroblasts compared with UVB-exposed cells (22.2 ± 2.7% vs. 36.7 ± 8.0%). Furthermore, pretreatment with EKGR prior to UVB exposure resulted in a significant increase in pro-collagen type I production (37,075.1 ± 7532.2 pg/mL) and a concomitant decrease in MMP-1 secretion (25,754.1 ± 4042.0 pg/mL) relative to UVB-exposed cells (26,845.8 ± 1454.6 and 39,910.8 ± 6035.1 pg/mL, respectively). To demonstrate formulation feasibility, EKGR was incorporated into an oil-in-water microemulsion, which exhibited concentration-dependent SPF enhancement. Collectively, these findings demonstrate the photoprotective efficacy of EPMC-rich EKGR and highlight its potential as a cosmeceutical ingredient for mitigating UVB-induced photo-senescence and skin aging, with an additional SPF boosting effect. To our knowledge, this study provides the first evidence of EKGR-mediated protection against UVB-induced cellular senescence in human dermal fibroblasts. Full article

In this study, we compared the effects of microwave-assisted extraction (MAE) and ultrasound-assisted extraction (UAE) on the total phenolic content, antioxidant activity, morphological characteristics, and tentative identification of bioactive compounds by LC-ESI-MS/MS in pomegranate seeds. We conducted a phytochemical characterization of the extracts by determining the total phenolic content and total flavonoids. Antioxidant activity was evaluated using ferric-reducing antioxidant power (FRAP) and free radical inhibition methods (DPPH and ABTS). Morphological characteristics were analyzed via scanning electron microscopy, UV-Vis and FTIR of the extracts were recorded. Additionally, the main bioactive compounds were identified using HPLC-MS. Our results demonstrated that MAE was the most efficient technique, yielding a higher content of total phenols (35.47 mg GAE/g), total flavonoids (14.44 mg CAE/g) and antioxidant activity (0.19 and 0.41 mmol TEAC/g, as determined by FRAP and ABTS, respectively). In terms of morphological characteristics, UAE induced more changes in the structure of the plant material compared to MAE. According to HPLC-MS analysis, the extract obtained using MAE notably contained coumaric acid, cyanidin, and quercetin, whereas the UAE extract included coumaric acid, cyanidin, kaempferol, and epicatechin. In conclusion, this study demonstrated that MAE is a more efficient method than UAE for extracting bioactive compounds. Pomegranate seeds may represent a potential source of these compounds for application in various industrial areas. Full article

Quality by design (QbD) is a systematic approach focused on achieving consistent, predictable quality based on predefined objectives. Unlike traditional methods, QbD prioritizes risk assessment and management, which significantly enhances the robustness of the analytical method. In this study, we initiated factor screening using a three-factor, two-level design to evaluate three independent variables: flow rate, pH, and mobile phase composition. To further investigate the interaction of these variables, we employed Central Composite Design (CCD). This allows us to apply response surface methodology to the Critical Analytical Attributes (CAAs), specifically retention time, peak area, and symmetry factor, by conforming to the method’s robustness. The combination of pregabalin and duloxetine hydrochloride (HCl) dosage form was determined using a straightforward, exact, specific, and accurate reverse-phase HPLC approach. The results showed retention times of 3.265 min and 4.318 min for duloxetine HCl and pregabalin, respectively. Pregabalin demonstrated linearity from 100 to 200 μg/mL (R² = 0.998), whilst duloxetine HCl demonstrated linearity between 20 and 120 μg/mL (R² = 0.997). Lower LOD values of 0.925 µg/mL and 0.853 μg/mL and LOQ values of 2.809 μg/mL and 2.587 μg/mL of pregabalin and duloxetine HCl, respectively, suggest good sensitivity for the technique. The drug content of the commercial formulation may thus be determined using the recommended method. This technique can be used for standard quality control studies to simultaneously estimate pregabalin and duloxetine HCl. The novelty of the present studies lies in the development of a robust RP-HPLC method for simultaneous estimation of pregabalin and duloxetine HCl using a systematic AQbD approach, enhancing robustness, reproducibility, and reliability, making it highly suitable for routine quality control and regulatory applications. Full article

Green synthesis is an eco-friendly, simple, and cost-effective process for the synthesis of metal nanoparticles from plant extracts that are rich in bioactive compounds. In the current study, the antioxidant potential and total soluble polyphenol content (TPC) of different parts of Ligusticum mutellina (L.) Crantz were evaluated using DPPH (2,2-diphenyl-1-picrylhydrazyl) and FRAP (ferric reducing antioxidant power) assays, and the results indicated that the seed extract was the most active plant part. HPLC analysis indicated the presence of phenolic compounds such as gallic acid, protocatechuic acid, and catechin, which may contribute to the reduction and stabilization of AgNPs. Silver nanoparticles (AgNPs) were synthesized from the aqueous seed extract of L. mutellina. The formation of nanoparticles was confirmed by UV–Vis spectroscopy, FT-IR analysis, powder X-ray diffraction (PXRD), and transmission electron microscopy (TEM). The UV–Vis spectrum indicated a surface plasmon resonance peak at around 411 nm, and PXRD analysis indicated an average crystallite size of around 13 nm. TEM analysis revealed predominantly spherical nanoparticles with an average size of 25.36 ± 10.76 nm. The synthesized AgNPs exhibited strong antibacterial activity against Gram-positive (Staphylococcus aureus and Bacillus subtilis) and Gram-negative (Escherichia coli and Pseudomonas aeruginosa) bacteria. Overall, the results demonstrate that L. mutellina seed extract represents an effective natural source of reducing and stabilizing agents for green nanoparticle synthesis and highlight the potential of the obtained AgNPs as environmentally friendly antimicrobial materials. Full article

Background/Objectives: Onion (Allium cepa) peems are an underutilized by-product rich in polyphenols. This study evaluated the physicochemical profile, and bioactive potential (antidiabetic, antimicrobial, antioxidant, and anticoagulant) of Moroccan red onion peels using integrated in vivo, in vitro, and in silico approaches. Methods: Moisture, pH, ash content, and mineral elements were determined, followed by phytochemical screening and three extractions: decoction E0, aqueous Soxhlet E1, and hydroethanolic Soxhlet E2 (70/30; ethanol/water, v/v). The measurement of polyphenols, flavonoids, and tannins was carried out using colorimetric methods, while the molecular profile was studied by high-performance liquid chromatography coupled to ultraviolet detection and electrospray ionization mass spectrometry (HPLC/UV-ESI-MS). Biological activities were determined using 2,2-diphenyl-1-picrylhydrazyl, ferric reducing antioxidant power, and total antioxidant capacity assays (in vitro antioxidant); microdilution (antimicrobial); prothrombin time and activated partial thromboplastin time (anticoagulant); and α-amylase/α-glucosidase enzymatic inhibition and oral glucose tolerance tests on normoglycemic rats. Also, acute toxicity was evaluated, and molecular interactions between these proteins and ligands (docking, molecular dynamics, and MM-PBSA) were analyzed. Results: Physicochemical analyses showed an acidic pH (3.06) and high ash content (15.21%), with the concentration of regulated elements remaining within FAO/WHO limits. The extractive content was between 6.90% E0 and 19.18% E2. The E1 extract had the maximum amount of total polyphenols (178.95 mg GAE/g); on the other hand, E2 was the richest in flavonoids by 121.43 mg QE/g. The HPLC/ESI-MS analysis of E0 revealed 20 compounds, among which flavonoids (84.93%) were predominant, with isorhamnetin (30.26%), followed by quercetin and its glycosylated forms. E1 showed the most potent antioxidant effects (IC₅₀ DPPH, 22.38 µg/mL, as that of ascorbic acid). The antibacterial activity of E0 was especially potent towards Enterobacter cloacae and Pseudomonas aeruginosa (MIC 75 µg/mL). A mild dose-dependent anticoagulant effect was seen. Antidiabetic activity was found to be outstanding: α-amylase (IC₅₀ 62.75 µg/mL) and α-glucosidase (IC₅₀ 8.49 µg/mL, stronger than acarbose) inhibitions were corroborated in vivo by a considerable decrease in the glycemic area under the curve. The molecular docking study in silico demonstrated strong molecular interactions, especially for quercetin 4′-O-glucoside with good binding energies. Conclusions: A. cepa peels from Morocco can be considered a safe plant matrix containing bioactive flavonoids with strong antioxidant and selective antimicrobial activities and promising antidiabetic effects, supported by molecular modeling. Full article

Background/Objectives: Atopic dermatitis (AD) is associated with gut dysbiosis linked to early-life antibiotic use and Staphylococcus aureus colonization. Gum Arabic (GA), a prebiotic, may modulate this dysbiosis and influence AD-related microbial balance. This study evaluated whether GA could support AD-associated probiotics-Lactobacillus casei, Bifidobacterium bifidum, and Bifidobacterium infantis-under amoxicillin- or azithromycin-containing conditions, examined the response of S. aureus under the same screening conditions, and developed GA-phospholipid-based semisolid carriers for topical application. Methods: Probiotic strains were cultured with 1–5% GA in the presence and absence of antibiotics, and viable cell counts were assessed. Sixteen topical formulations containing propylene glycol or isopropyl myristate in a hydrogenated phosphatidylcholine base were prepared and screened for rheological properties and galactose release using in vitro release testing (IVRT) and HPLC-UV. Results: GA at 1–2% concentrations promoted probiotic growth in antibiotic-free conditions. GA preserved B. infantis viability under azithromycin exposure in this in vitro screening model. For S. aureus, numerical CFU differences were observed between antibiotic-only and GA-containing conditions; however, the present screening design was not intended to determine antibiotic interaction outcomes. Formulations F14 (2% GA + 7% IPM) and F15 (3% GA + 7% IPM) exhibited optimal spreadability. IVRT showed that 6 h cumulative galactose release varied by formulation (F6 > F10 > F14 > F15). Conclusions: GA demonstrated dose-dependent prebiotic activity and preserved B. infantis viability under azithromycin exposure in this in vitro screening model. For S. aureus, the observed CFU differences between antibiotic-only and GA-containing conditions should be considered exploratory only and do not allow for conclusions regarding interference with antibiotic efficacy. Optimized GA-HPC systems with suitable rheological and release characteristics represent promising candidates for further preclinical investigation. Full article

Aim: This study investigates how Buyang Huanwu Decoction (BHD) protects against cerebral ischemic damage by targeting the NLRP3/Caspase-1 pathway. Methods: The fingerprint of BHD was analyzed by HPLC-UV. Migratory chemicals in BHD-containing cerebrospinal fluid (BHD-CCSF) were analyzed by ultra-performance liquid chromatography-quadrupole-time of flight-mass spectrometry (UPLC-Q-TOF-MS). The effects of BHD on the NLRP3/Caspase-1 pathway, IL-18 and IL-1β levels in oxygen and glucose deprivation/reperfusion (OGD/R) cells were assessed by Western blot and ELISA. Cerebral infarction severity in permanent middle cerebral artery occlusion (pMCAO) mice was assessed by mNSS scores and staining. Protein and mRNA levels of the NLRP3/Caspase-1 pathway and inflammatory factors (IL-18, IL-1β) were measured. Results: BHD-containing serum (BHD-CS), BHD-CCSF, and Calycosin (Cal) reduced NLRP3, Caspase-1, ASC, GSDMD proteins, IL-18 and IL-1β in OGD/R cells. In pMCAO mice, BHD decreased pathway-related proteins and mRNA and inflammatory factors and alleviated brain injury. Conclusions: BHD ameliorates cerebral ischemia by inhibiting the NLRP3/Caspase-1 pathway, thereby suppressing pyroptosis and inflammation. Full article

Removal of micropollutants from wastewater is currently drawing a lot of attention in the field of municipal wastewater treatment plants. Firstly, this is because of their unpredictable and potentially damaging fate in the environment, and secondly, due to newly established requirements in the relevant European Union Directive (EU) 2024/3019. This article assesses coagulation, oxidation with chlorine dioxide, sand biofilter, and their combinations as potentially cheaper and sustainable alternatives to well-established, but more expensive methods. For the experiments, citalopram, carbamazepine, and diclofenac were chosen as representatives of micropollutants. Removal efficiencies were evaluated using HPLC, COD, and absorbance UV/VIS at different wavelengths. The demand for chlorine dioxide was assessed using the chlorophenol red method. Owing to analytical limitations, the concentrations examined were in mg/L, which significantly exceeds actual concentrations found in wastewater. The application of stand-alone chlorine dioxide oxidation exhibited the best performance as it sufficiently removed citalopram and diclofenac. On the contrary, biodegradation was found to be the least efficient method, as none of the compounds tested were sufficiently removed in a short period of time. However, the results may be partially biased owing to high concentrations of the micropollutants assessed. In the following stage of the research, the evaluation of transformation products is desired to prevent such potentially harmful chemicals from entering the environment. Full article