Citrus peel wastes are valuable sources of phenolic compounds (PCs); however, the industrial essential oil extraction exposes the peels to heat, which may significantly affect their concentration, stability, composition, and biological activities. This study aimed to evaluate the impact of thermal processing associated with prior essential oil recovery on the phenolic metabolome and bioactivity of lemon, mandarin, and orange peels. Aqueous-ethanol extracts were prepared from crude peels (CPEs) and heat-treated residues remaining after the essential oil extraction (HPEs) of citrus peel wastes. The molecular-network-assisted LC-MS/MS analysis identified a total of 120 annotated metabolites, predominantly 98 phenolics, and showed a marked reduction in phenolic diversity after heat treatment. In CPEs 59, 55, and 44, phenolics were detected in lemon, mandarin, and orange, whereas phenolics in 43, 49, and 43 were noticed in their corresponding HPEs, respectively. CPEs showed stronger antioxidant activity, with lower IC50 values in the DPPH assay (38.72, 60.38, and 70.77 mu g/mL) and NO inhibition assay (61.54, 90.32, and 117.50 mu g/mL) compared with HPEs. The extracts also exhibited significant anti-inflammatory activity through selective inhibitory effects against the COX-2 enzymes. Overall, thermal treatment during essential oil extraction markedly altered the phenolic composition and reduced the biological activity of citrus peel extracts. These findings demonstrate that processing conditions critically influence the functional value of citrus peel wastes.

Microalgae are increasingly recognized as sustainable bioresources with applications in food, feed, nutraceuticals, and aquaculture. Among them, Spirulina (Limnospira platensis) and Microchloropsis gaditana (formerly Nannochloropsis gaditana) stand out due to their rich biochemical composition with industrial applications and high market value. This study separately investigates the seasonal variability in the biochemical composition of L. platensis cultivated in open raceway ponds and M. gaditana grown in closed photobioreactors. The results suggest a response to seasonal changes. L. platensis exhibited higher protein content in spring (47.82%) and summer (45.48%), while carbohydrate accumulation peaked in winter (50.85%). M. gaditana showed increased lipid (21.67%) and carbohydrate (14.56%) content in autumn compared to winter and summer, but with lower variations in response to seasonal environmental changes. This microalga exhibited higher FA 20:5 n-3 (EPA) levels in winter compared to summer. The dominant carbohydrate residue in both microalgae, present throughout all seasons, was glucose, which could be associated with the presence of beta-glucans, in addition to glycogen, reinforcing their bioactive potential. Pigment composition was also seasonally influenced in L. platensis, which accumulated more chlorophyll and phycocyanin in spring and summer, whereas M. gaditana showed elevated levels of neoxanthin in autumn and winter, and violaxanthin in spring. Although limited by a single sampling per season and the different cultivation systems size, this study provides preliminary observations of seasonal variations in biomass composition, highlighting that biochemical composition varies with season in a species-specific manner. Understanding these variations improves our knowledge of microalgal biochemical plasticity under outdoor conditions.

Porcine blood is a major slaughterhouse by-product and a sustainable source of high-quality proteins with potential food and nutraceutical applications. This study valorized porcine whole blood (WB, 6.7 ± 0.1% protein) and red cell fraction (CF, 50.4 ± 0.2% protein) through alcalase hydrolysis, generating hydrolysates (WBH and CFH) with bioactive and techno-functional properties. Optimal hydrolysis conditions, defined as enzyme-to-substrate (E/S) and incubation time yielding the highest degree of hydrolysis (DH) with cost-effective enzyme usage, were 1% E/S for 4 h (WBH) and 2.5% E/S for 4 h (CFH). WBH showed a higher DH (59.5 ± 2.6%) than CFH (30.8 ± 3.3%). Antioxidant assays revealed higher ABTS activity in CFH (14.1 vs. 11.1 mg ascorbic acid equivalents/g, p < 0.05), while both exhibited similar ORAC values (166.8–180.2 mg Trolox equivalents/g, p > 0.05). After simulated gastrointestinal digestion, ABTS activity was preserved, whereas ORAC decreased (~40%). ACE inhibitory activity was also pronounced, particularly in CFH (IC50 = 59.5 µg protein/mL), but digestion converged values between hydrolysates (118–135 µg protein/mL). Techno-functional tests showed moderate emulsifying activity (~40%), with CFH displaying markedly higher oil absorption (4.79 vs. 1.31 g oil/g). Considering the limited information on porcine blood hydrolysates under gastrointestinal conditions, these findings provide new insights into their stability and support their potential as multifunctional ingredients for health-promoting foods and functional formulations.

The focus on by-product valorization in the food industry, particularly from the carob pod, underscores a commitment to sustainability and resource efficiency. This fruit, sourced from the leguminous evergreen carob tree (Ceratonia siliqua L.), is renowned for its adaptable flavour and nutritional value, in Mediterranean regions such as Portugal. Its production yields significant by-products, presenting environmental challenges when not managed efficiently. Innovative approaches, including integral carob flour production, aim to optimize utilization while minimizing waste and energy consumption. This study repurposed carob waste to produce novel, value-added ingredients like carob syrup, by thermal hydrolysis of integral carob flour using water at 1:3 solid-to-liquid ratio - obtaining up to 50 % solubility yield. The resulting syrup exhibited 72 % degrees Brix, a melting temperature (Tm) of approximately 130 degrees C and predominantly viscous behavior with minimal elastic (solid-like) response. Lastly, the syrup was incorporated into a carob-based brigadeiro, replacing conventional glucose-fructose syrup. Simulated gastrointestinal digestion revealed enhanced bioaccessibility of sugars and phenolics, and increased antioxidant activity during the intestinal phase. Despite sugar availability, the prebiotic activity of the syrup decreased when embedded in the brigadeiro matrix, potentially due to interactions with polyphenols or organic acids. Cytotoxicity and permeability assays confirmed safety at <= 0.5 % (w/v) and supported intestinal barrier integrity. These findings support the use of integral carob flour for producing multifunctional ingredients, contributing to circular economy models while meeting consumer demands for healthier, sustainable food products.

Recently, several studies have investigated strategies to reduce the antagonistic ripening effect of 1-methylcyclopropene (1-MCP). In this study, we examined how abscisic acid (ABA) influences ethylene production and fruit ripening in 'Rocha' pears treated with 1-MCP. To explore potential mechanisms for reactivating ripening, 'Rocha' pears treated with 1-MCP were exposed to 50 mg mL-1 of ABA and stored at 20 +/- 2 degrees C for 15 days. Typical ripening indicators, such as firmness, skin colour, ethylene and aroma volatile production, sugar content, and the expression of ethylene-related enzymes and receptors, were measured throughout the 15 days of storage. Multivariate and clustering analyses showed that the treatment countered the effects of 1-MCP, with ABA significantly increasing ethylene production, fruit softening, yellowing, simple sugar accumulation, and the emission of specific ripening volatile organic compounds, especially acetates, compared to pears exposed only to 1-MCP. Furthermore, ABA boosted the expression of ethylene biosynthesis genes (PcACS, PcACO) and the receptor gene (PcETR2), confirming its role in enhancing ethylene metabolism. Overall, the findings indicate that ABA influences metabolic pathways closely associated with ripening. This research provides new insights into the ripening mechanisms of pears and highlights the regulatory interaction between ABA and ethylene.

Pectin is a naturally occurring biopolymer extensively used for applications in the pharmaceutical, biotechnology, and food industries, and is abundantly present in lemon by-products. Although lemon peels represent a highly promising raw material, the structure of pectin is strongly influenced by extraction and drying processes, and the resulting attributes remain insufficiently understood. The present study systematically investigates the impact of conventional hot acid extraction using three different acidifying agents (citric, sulphuric, and hydrochloric) in combination with two drying techniques (oven-drying and freeze-drying) on the physicochemical, structural, thermal, and viscosity-shear rate properties of pectin obtained from lemon by-products (Citrus limon, Portuguese Eureka variety) following the prior recovery of bioactive compounds (essential oils and phenolic compounds). The results demonstrated that citric acid extraction followed by freeze-drying yielded the highest pectin recovery, at approximately 26.7%, highlighting the suitability of this approach for efficient by-product valorisation. Oven-dried pectins exhibited higher moisture contents (8.5-10%) and lower lightness values (L* = 57.02-65.67), indicating darker colouration compared to freeze-dried pectins (L* = 78.82-83.75). All extracted pectins presented a degree of esterification (DE ≥ 50%), classifying them as high-methoxyl pectins. The galacturonic acid (GalA) content ranged from 37.6 to 48.9% for oven-dried samples and increased to 44.1-58.6% for freeze-dried samples. Furthermore, pectin obtained from lemon by-products exhibited a well-defined structural organisation and enhanced thermal stability, especially for freeze-dried pectin samples, and suitable rheological properties, with no statistically significant variations observed between different acids or drying conditions, supporting its technological suitability for applications in the food, cosmetic, and pharmaceutical industries.

Protein hydrolysates derived from insects have been explored in recent years for their bioactive functionalities, namely antioxidant and antihypertensive activities, although the investigation of in vivo effects remains scarce. The present study aimed to assess the effects of dietary inclusion of black soldier fly (Hermetia illucens) larvae (BSFL) hydrolysates on nutritional status and cardiac biomarkers of adult Beagle dogs. Three complete diets were used: a control diet containing 3% shrimp hydrolysate (D_Shrimp), and two experimental diets in which the shrimp hydrolysate was replaced by BSFL hydrolysates obtained through the enzymatic hydrolysis with either ALCALASE 2.5L (D_BSFL-A) or Corolase PP (D_BSFL-C). Two-bowl tests were performed to determine palatability by pairwise comparison between all diets. A feeding trial designed according to four 3 x 3 Latin squares with twelve adult Beagle dogs, three periods of 28 days each, and the three complete diets was performed. A cardiac evaluation was performed to all dogs before the trial. Diet digestibility, fecal characteristics and metabolites, skin and coat quality, and cardiac biomarkers were assessed at the end of each period. No significant differences were observed on first approach, first-taste, and intake ratio. Experimental diets increased organic matter (P = 0.038) and crude protein (P = 0.048) digestibility compared to control, and dry matter digestibility was greater (P = 0.048) for D_BSFL-A than D_BSFL-C. No significant differences were observed in serum levels of C-reactive protein, cardiac troponin I, and NT-proBNP among diets, but dogs fed D_BSFL-A tended to have a lower (P = 0.086) serum insulin-like growth factor 1 level compared to dogs fed D_BSFL-C. Although all echocardiographic measurements remained within normal reference ranges, experimental diets promoted a lower (P = 0.030) interventricular septum diameter in diastole compared to the control, and a lower pulmonary vein diameter was noted for D_BSFL-C compared to D_BSFL-A (P = 0.049). Also, dogs fed D_BSFL-C showed a tendency to lower blood pressure (P = 0.102), serum angiotensin converting enzyme (P = 0.092), and left atrial-to-aortic root ratio (P = 0.062) compared to dogs fed D_BSFL-A. In conclusion, diets containing BSFL hydrolysates are well accepted and digested by dogs and might influence mechanisms involved in blood pressure regulation and potentially modulate cardiac function, although further studies are warranted.

Underutilized by-products from the walnut-oil industry, namely walnut oilcake (WOC) and walnut oil dregs (WOD), were evaluated for their nutritional composition, phenolic compound profile and digestive behaviour, as well as bioactive properties (antioxidant, antimicrobial, anti-inflammatory, cytotoxic and prebiotic activities). WOC was rich in protein (38.1 g/100 g) and dietary fiber (32.6 g/100 g), while WOD presented high fat (46.8 g/100 g) and carbohydrate content (20.9 g/100 g). Glansreginin A was the predominant phenolic compound in both matrices. Following in vitro digestion using the INFOGEST protocol, higher overall polyphenol bioaccessibility was noticed in WOD (78%) compared to WOC (15%). Bioaccessible fractions exhibited higher antioxidant activity than the undigested samples. Glansreginin A was detected only on the cellular apical compartment suggesting the absence of transport across Caco-2 cells. After in vitro digestion, the non-bioaccessible fractions enhanced the growth of Lactobacillus and Bifidobacterium strains, in some cases surpassing fructooligosaccharides, a standard prebiotic. These findings support the valorisation of walnut by-products as functional ingredients, also contributing to sustainable food systems.

Pterospartum tridentatum (L.) Willk. (carqueja) is widely used in traditional medicine and culinary practices in the Iberian Peninsula; however, most studies have focused on its flowers, while its stems remain largely unexplored, despite representing a significant proportion of the plant biomass. This study aimed to evaluate the potential of P. tridentatum stems as a source of bioactive compounds using different extraction methodologies. Aqueous, hydroethanolic, ultrasound-assisted extraction (UAE) and pressurized liquid extraction (PLE) were applied, and the resulting extracts were characterized in terms of their extraction yield, protein and carbohydrate content, phenolic composition, antioxidant capacity, antimicrobial activity, and cytotoxicity in HaCaT and Caco-2 cell lines. Phenolic profiling by LC-ESI-QqTOF-HRMS tentatively identified 37 compounds, mainly corresponding to flavonoid and isoflavonoid glycosides, with genistein derivatives representing the dominant constituents across all extracts. Although extraction yields differed among methods, phenolic profiles were broadly similar. UAE and PLE extracts showed slightly higher antioxidant activity, while antimicrobial activity was limited, with only moderate inhibition observed against Staphylococcus epidermidis and Malassezia furfur. Additionally, cytotoxicity assays indicated low toxicity. Overall, the results demonstrate that P. tridentatum stems represent a promising yet underutilized biomass source of phenolic compounds with antioxidant potential and low cytotoxicity under the tested in vitro conditions.

The leaves of Quercus ilex, rich in phenolic compounds and dietary fibre, represent a promising source of bioactive substances with potential health benefits. In vitro human colonic fermentation was performed to evaluate the prebiotic effect of ground Q. ilex leaves. Prior to fermentation, (-) epicatechin, catechin, and rutin were identified as the predominant phenolic compounds that reached the colon after gastrointestinal digestion, with (-) epicatechin present at the highest concentration (74 mg/100 g DM). Among then, rutin undergoing the most extensive metabolization during fermentation, decreasing from 22 mg/100 g DM at baseline to below the detection limit after 48 h of fermentation. Compared with negative control, Q. ilex leaves increased the abundance of Bifidobacterium spp. and Prevotella spp. by 46% and 40%, respectively, after 24 h of fermentation, whereas fructooligosaccharides (FOS) increased these bacterial groups by 31% and 16%, respectively. In addition, ground Q. ilex leaves enhanced the production of short-chain fatty acids (SCFAs), particularly acetate and propionate, which reached their highest concentrations after 12 h of fermentation. Moreover, Q. ilex leaves tend to maintain a Firmicutes:Bacteroidetes ratio closer to 1 than FOS throughout fermentation (1,31 vs. 1,82), suggesting a more balanced gut microbiota composition. Overall, the observed effects of Q. ilex leaves support their potential use as a natural and currently undervalued source of bioactive compounds with prebiotic properties. Considering the growing interest in sustainable, low-impact and locally sourced food ingredients, Q. ilex by-products may represent a promising alternative for the development of new functional foods and nutraceutical formulations designed to promote gut health.

Fucoidan (FPS), a sulfated polysaccharide isolated from brown algae with a molecular weight ranging approximately from 5 to 200 kDa, exhibits diverse bioactivities, yet its high molecular weight (HMW) restricts topical bioavailability. This study explored the molecular-weight-dependent transdermal behavior of FPS and its underlying interaction mechanisms with the skin barrier. To address this, FPS fractions (6 to 103 kDa) were prepared via controlled oxidative degradation. In vitro permeation studies combined with Confocal Laser Scanning Microscopy (CLSM) visualization revealed a critical molecular weight threshold of approximately 11 kDa. HMW-FPS were mainly retained on the skin surface, whereas low molecular weight FPS (LMW-FPS, ≤11 kDa) penetrated into the viable epidermis and dermis. ATR-FTIR spectroscopy was employed to elucidate the underlying mechanism, which revealed that LMW-FPS overcomes the skin barrier through synergistic structural modulations: (1) it enhances intercellular lipid fluidity, accompanied by a reduction in CH2 stretching vibration intensity; (2) it induces conformational changes in keratin via direct electrostatic interactions, promoting the transition from α-helices to β-sheets. Furthermore, histological evaluation confirmed that FPS treatment caused no obvious skin irritation. These findings demonstrate that LMW-FPS acts as a safe, reversible modulator of the stratum corneum (SC) barrier, providing a promising strategy for the design of polysaccharide-based transdermal delivery systems.

Phenolic compounds are recognized for antioxidant, anti-inflammatory, and antidiabetic properties. Lentils are abundant in these compounds, yet comparative studies across varieties and intestinal cells effects remain limited. This study evaluated the nutritional composition, phenolic profile, antioxidant activity, and α-glucosidase inhibition of four lentil types in raw and cooked forms. L01 had the highest bioactive potential and was the only one selected for further evaluation in Caco-2 cells under basal and IL-1β-stimulated conditions. Cooking altered composition, leading to higher protein, fiber, and carbohydrates, reducing tannins, and maintaining high phytic acid in some varieties. L01 consistently showed the highest phenolic content, antioxidant activity, and enzyme inhibition. Its phenolic profile was dominated by kaempferol derivatives, (epi)catechin, and procyanidins, with cooking increasing monomeric catechins but reducing procyanidin oligomers. At noncytotoxic levels, L01 extracts reduced IL-6/8 secretion in stimulated cells, with stronger effects from raw extracts. Additionally, they decreased expression of inflammatory markers (IL-6/8/1β) while increasing expression of metabolic regulation- and barrier-related genes (Peroxisome proliferator-activated receptor-γ, Sirtuin 1, Occludin, and Cadherin-1). Only raw extracts significantly enhanced Heme oxygenase-1 expression under stimulation. These findings highlight compositional differences beyond tegument color and support further investigation of phenolic-rich lentils as potential functional food ingredients targeting gut and metabolic health.

Background/Objectives: Carotenoids are bioactive pigments with well-established antioxidant and immunomodulatory properties, yet their impact on gut microbiota remains poorly understood from a chemical standpoint. This study explores how carotenoid structure and gastrointestinal stability shape microbial responses combining in vitro fermentation with bioinformatic analyses. Methods: Individual carotenoids (beta (β)-carotene, lutein, lycopene) and combined carotenoids, as well as algal-derived extracts were subjected to 48 h in vitro fermentation, and microbial composition and activity were assessed through sequencing and computational analysis. Results: β-carotene and lycopene promoted acid-tolerant taxa such as Escherichia-Shigella, whereas lutein, due to its higher polarity, supported more transient fluctuations. Mixtures and algal carotenoids exhibited synergistic effects, sustaining beneficial genera including Bifidobacterium and Bacteroides and promoting structured ecological trajectories. Conclusions: These findings provide a chemistry-driven perspective on how carotenoids act as modulators of microbial ecosystems, with direct implications for the formulation of carotenoid-enriched functional foods and dietary interventions.

This study evaluates the influence of harvesting methods and seasonal variability on the physicochemical and antioxidant properties of Tetragonisca fiebrigi honey produced in the tropical dry forest of Bolivia. Despite the growing interest in stingless bee honey, studies addressing the combined effects of seasonality and collection practices in this region remain scarce. Honey samples were collected during winter and spring using three approaches: conventional, optimized (based on good manufacturing practices), and direct racking from natural nests. Physicochemical parameters (pH 4.60-6.15; moisture 28-34%; water activity 0.69-0.75) and sugar composition (glucose 10.60-29.03 g/100 g; fructose 9.01-21.97 g/100 g; sucrose 0.70-3.23 g/100 g) showed variability primarily associated with season rather than harvesting method. Bioactive compounds exhibited a marked seasonal effect, with higher total phenolic content (up to 11.03 mg GAE/100 g), flavonoids (up to 23.08 mg QE/100 g), and antioxidant capacity (DPPH up to 1.33 mol TE/100 g; ORAC up to 25.93 mol TE/100 g) in spring samples. Multivariate analysis (PCA) revealed that honey variability is structured along bioactive and physicochemical axes, with samples obtained using the optimized method showing reduced dispersion and greater compositional consistency. These results indicate that while seasonality governs the compositional and functional properties of T. fiebrigi honey, improved harvesting practices contribute to reducing variability and enhancing product standardization. This study provides one of the first comprehensive datasets on Bolivian stingless bee honey and highlights its potential as a functional food, supporting the development of species-specific quality criteria and sustainable meliponiculture in tropical dry forest ecosystems.

Pulsed electric field (PEF) technology was used as a green extraction method for the simultaneous recovery of starch and polyphenols from acorns using water as extraction solvent, and the gelatinization properties of the extracted starch were compared with those of a starch obtained via alkaline extraction. Results showed that the electric field intensity had a significant negative quadratic effect on the antioxidant activity and hydrolyzable tannin content, whereas it had a positive quadratic effect on the starch yields. Time had no significant effect on the starch yields but had a negative quadratic effect on the phenolic and hydrolyzable tannin contents and antioxidant activity. The extraction condition that allowed the highest starch and polyphenol contents with the greatest antioxidant activity was 0.1 kV/cm for 63.3 µs, with a desirability of 87%. In comparison to alkaline extraction, PEF increased tannin extraction by 11-fold, highlighting its potential to produce tannin-rich extracts suitable for leather manufacture. Although the starch yield was 30% lower than that obtained via alkaline extraction, the PEF-isolated starch is a cleaner and safer ingredient for human consumption. The alkaline starch had a significantly higher solubility than the PEF-isolated starch, but both had similar swelling power.

Wheat processing generates large volumes of co-products, particularly wheat bran (WB) and wheat germ (WG), which remain underutilized despite their high content of dietary fiber, phenolic compounds, bioactive peptides, and lipophilic antioxidants. Although their composition and processing have been widely investigated, an integrated and application-oriented evaluation of these fractions remains limited. This review provides a structured and critical analysis of WB, raw and defatted WG, and wheat germ oil (WGO), linking composition, processing strategies, and functional performance within a unified framework. Conventional and emerging technologies, including enzymatic hydrolysis, fermentation, thermomechanical treatments, and supercritical CO2 extraction, are discussed in terms of selectivity, impact on techno-functional properties, and scalability. An evidence-grading approach is introduced to distinguish bioactivities supported by chemical assays, cell-based models, animal studies, or human data, enabling a more rigorous interpretation of health-related effects. Across applications, these co-products have been incorporated into food systems and related sectors, primarily showing improvements in nutritional composition, oxidative stability, and product performance under experimental conditions. However, translation to an industrial scale remains constrained by techno-economic limitations, regulatory requirements, and stability challenges. This work highlights the need for integrated processing strategies aligned with industrial feasibility to support the development of sustainable cereal biorefineries.

Transforming seafood by-products into valuable ingredients presents a sustainable approach to advancing food innovation. This study aimed to characterize sardine protein hydrolysates (SPH), obtained by enzymatic hydrolysis, and evaluate their nutritional profile and multifunctional bioactivities, including the effect of enrichment with rosemary extract. Sardine by-products were hydrolyzed with alcalase, and the resulting SPH were analyzed for proximate composition, amino acid profile, peptide size distribution, lipid composition, and mineral content. Bioactivity was assessed through in vitro cell-based assays in Caco-2, RAW 264.7, and 3T3-L1 cells, testing cytotoxicity, oxidative stress modulation, lipolysis inhibition, and TNF-α expression. SPH contained 57.1

Global plastic production is expected to continue rising in the next few decades, with packaging accounting for roughly one quarter of the total volume of plastic and intensifying interest in biodegradable alternatives such as polylactic acid (PLA). One approach towards bioplastic production is lactic acid fermentation. Lactic acid (LA) can be produced from waste and by-products, such as food and agricultural waste, and then polymerised into PLA. This review provides an overview of LA production from waste-derived feedstocks, covering substrate composition, pretreatment and hydrolysis strategies, fermentation modes, and downstream operations, and highlights key performance indicators (yield, final LA concentration, and volumetric productivity). This review examines technological bottlenecks associated with waste heterogeneity, mixed sugar utilisation, inhibitor formation, pH and temperature control, contamination risks, high cost, and complexity of LA recovery and purification. Recent advances in pre-treatment, robust or engineered microbial strains, mixed microbial cultures, process integration, and intensified downstream schemes are also discussed to increase productivity and purity while reducing energy and chemical inputs. Finally, techno-economic assessment studies on waste-based PLA are synthesised, showing that both economic and environmental performance depend on feedstock logistics, process configuration, and end-of-life options for PLA products.

Photoaging, driven by chronic ultraviolet (UV) exposure, accelerates skin aging through inflammation, collagen degradation, and oxidative stress. This study presents a novel and sustainable anti-photoaging formulation, Nano Citrus Cream (NCC), based on chitosan-pectin (CHI-PEC) nanoparticles loaded with a polyphenol-rich grapefruit extract and orange essential oil. The optimized nanoparticles (CH-PEC, 1:1) exhibited high encapsulation efficiency (up to 87.9%), nanoscale size (49-101 nm), and colloidal stability (+30 to +54 mV). When incorporated into a topical cream, NCC showed shear-thinning rheology, controlled polyphenols release, and was well-tolerated in vivo. In a UVB-induced photoaging rat model, NCC significantly reduced erythema, scaling, and histopathological alterations, outperforming non-encapsulated extract treatments. Biochemical analyses demonstrated downregulation of MMP3, TSP1, and LMN332, key markers associated with extracellular matrix degradation, inflammation, and basement membrane disruption. LC-ESI-HRMS profiling identified 71 bioactive compounds from phenolic, lipid-derived and terpenoid classes, several of which exhibited favorable predicted skin permeability. Molecular docking was applied as an exploratory, hypothesis-generating tool to assess potential interactions between selected skin-permeable metabolites and photoaging-realted enzymes, without assigning compound-specific dominance. Overall, the results support a nano-enable, multicomponent mode of action underlying the anti-photoaging efficacy of NCC. This work highlights NCC as a safe, biodegradable, and sustainable cosmetic formulation, while demonstrating the potential of nanotechnology-based delivery systems for valorizing citrus by-products in skin health applications.

Protein hydrolysates are increasingly used in feed, underscoring the need for analytical tools for the rapid, reliable determination of total protein. This study revisited a merging-zones flow-based spectrophotometric method for minimizing reagent consumption and employs the Biuret reaction for the quantification of total soluble protein in by-products hydrolysates. The analytical method was optimized across different physical and chemical parameters such as: flowrate, reactor length, sample injection volume and reagents concentration. The use of different matrices relevant in the hydrolysis processes (acetate, phosphate, and hydrochloric acid) showed no significant interference (<10%) on the method's performance. Under optimal conditions, the method quantified hydrolysate protein within a dynamic range of 0.100-2.00 mg mL(-1) , with LOD and LOQ of 0.069 and 0.290 mg mL(-1) , respectively. Analyses of hydrolysates showed no significant differences compared with the reference method (<10%) while reducing analysis time from 30 min to 3 min per sample (triplicate). The method's greenness, assessed by AGREE software, showed an improved score, from 0.58 to 0.76. The optimized protocol provides a fast, robust, and greener approach for monitoring protein hydrolysates in the food industry.