The composition and regiospecific distribution of fatty acids in TAG molecules is species-specific and varies within species. These are very important in determining milk fat's nutritional properties, impact on tissue metabolism, and functional properties affecting human health. TAG was hydrolysed by pancreatic lipase, and fatty acids were isolated by TLC separation and finally analysed using GC. Different trends were observed for the saturated and unsaturated fatty acid profiles among different species and within species, and the nutritional indices also showed significant differences between cow and buffalo milk fat. Strong species-wise variation in the sn-2 and sn-1(3) fatty acid distribution was often observed by analyzing data from across the whole year (intra-positional distribution). The mol % of saturated fatty acids at sn-2 positions was significantly (p≤0.05) higher in cow milk fat (73.49±0.53), while it was significantly (p≤0.05) higher in the sn-1(3) position in buffalo milk fat (71.23±1.61). On the contrary, the mol% of unsaturated fatty acids (UFAs) was significantly (p≤0.05) higher in the sn-2 position in buffalo milk fat (33.17±1.16) than in cow milk fat (26.51±0.84); the mol% of UFAs was significantly (p≤0.05) higher in the sn-1(3) position in cow milk fat (31.14±0.1) than in buffalo milk fat (25.33±0.59). A PCA analysis indicated that the sn-2-positional fatty acids were unique to each species. The MUFA and conjugated linoleic acid content were highest in the sn-2 position in buffalo milk. In buffalo milk fat, a higher amount of SFAs were converted to MUFA and PUFA, representing a higher Δ-9 desaturase index at the sn-2 position in TAG. PCA analysis also revealed differences in the sn-2- and sn-1(3)-positional distribution of TAG fatty acids in milk fat among different species. The Euclidean distance matrix of the quantitative data was also employed to group and categorize the fatty acids using heatmaps. The observed variations among various species revealed the significance of positional analysis in the characterization of milk fat from multiple species.

Grape seed extract (GSE), rich in polyphenolic compounds such as catechins and proanthocyanidins, exhibits strong antioxidant potential. This study explored the protective effects of GSE on oxidative stress resistance and longevity in Caenorhabditis elegans. Worms were treated with 100 and 500 μg/mL GSE from L1 to adulthood. In vitro assays confirmed enhanced DPPH and ABTS radical scavenging. In vivo, 500 μg/mL of GSE significantly increased median lifespan by 16.67%, improved thermotolerance by 17.65%, and promoted body growth and motor activity. Fertility remained unaffected, suggesting non-toxic longevity benefits. Biochemical assays showed elevated SOD, CAT, and GSH-Px activities, with reduced MDA levels, indicating enhanced antioxidant defenses. RT-qPCR revealed upregulation of skn-1, daf-16, sod-3, gst-4, and hsp-16.2, pointing to the activation of stress response pathways. Notably, GSE also increased the expression of set-2, suggesting epigenetic regulation via H3K4 methylation. GC-MS-based metabolomics revealed that GSE-treated worms exhibited altered amino acid metabolism, including elevated levels of glutamine, glutamate, and glycine—precursors for glutathione synthesis—alongside reduced pyruvate and succinate, indicating improved mitochondrial function and reduced metabolic stress. Increased norvaline and phenylalanine levels further suggested modulation of nitrogen metabolism and antioxidant pathways.

Collectively, these findings demonstrate that GSE confers significant anti-aging effects in C. elegans through coordinated activation of antioxidant enzyme systems, transcriptional upregulation of stress-responsive genes, epigenetic modulation, and metabolic reprogramming. This study provides mechanistic insights into GSE’s bioactivity and supports its potential as a dietary polyphenol for managing oxidative stress-related aging and metabolic decline.

Introduction: Milk-based oil is a bioactive and nutrient-rich food item, containing fat-soluble vitamins, essential fatty acids, and Rumenic Acid (RA). However, high cholesterol and saturated fatty acid levels make it challenging to consume. Some indices based on fatty acid composition have been defined to address this issue:
HPI=(∑(USFAs))/(∑C12:0+C16:0+(4×C14:0))
AI=(∑C12:0+C16:0+(4×C14:0))/(∑(MUFAs+PUFAs))
TI=(∑C14:0+C16:0+C18:0)/(0.5∑MUFAs+0.5∑PUFAs(n6)+3∑PUFAs(n3)+n3/n6)
Method and materials: In this cross-sectional study, the nutritional indices of 12 sheep and 12 cows’ butter oil were assessed and compared. The fatty acid composition of the oils was determined using gas chromatography after direct trans-esterification. The calculations of the HPI, AI, and TI were carried out using statistical tests in SPSS software.
Results: The study revealed that cow's butter oil had a higher HPI mean of 0.40±0.03 than sheep's butter oil, which had a mean of 0.35±0.28 (P <0.001). Additionally, the cow's butter oil had a lower AI mean of 1.96±0.26 compared to the sheep's butter oil, which had a mean of 2.42±0.33 (P <0.001). Lastly, the TI was 2.78±0.26 in cow's butter oil vs. 2.96±0.25 in sheep's butter oil (P <0.064).
Conclusion: It appears that, based on the indices for assessing the nutritional value and consumer health benefits of animal fat, cow's butter oil may be better than sheep's butter oil for human nutrition. However, further clinical trial studies are recommended to investigate the effects of butter oil on the Partial Thromboplastin Time (PTT) test and Atherogenic Indices of Plasma in both animal and human models.

In lipidomic research, the accurate quantification and identification of fatty acids in biological samples present methodological and analytical difficulties. From extraction to analysis, the choice of reagents, precision of derivatization, and analytical parameters all influence researchers’ proper understanding of the lipid profile. This is particularly relevant in studies of modern Western diets, where robust analytical methods are critical for assessing human nutritional health. This study aims to optimize key steps in fatty acid extraction, derivatization, and chromatographic analysis. Initial testing was conducted on chicken liver samples, with the intention of applying the optimized method to human samples in subsequent work. By testing different reagent concentrations, we demonstrated that a 46.5% solution of H₂SO₄ in methanol resulted in the most efficient methylation of fatty acids into fatty acid methyl esters (FAMEs), which was measured as higher peak intensities. In contrast, chromatographic separation was primarily influenced by column temperature gradients and injection parameters. The results indicate that a GC oven program starting at 100 °C, with a ramp of 7 °C/min to 240 °C, most effectively separated FAME peaks. Additionally, the use of a phosphate buffer at pH 5.8 during extraction was found to marginally improve peak resolution and amplitude compared to pH 7.0. All results were validated by assessing repeatability through repeated injections, inter-rater reliability, and variation in injection volumes. The novelty of our method lies in its application to soft tissue samples and in the systematic optimization and integration of multiple parameters, such as buffer composition, acid concentration for methylation, and GC/MS settings, which result in a robust, reproducible protocol suitable for high-quality analysis of fatty acids in complex biological matrices.

The increasing scientific interest in Verbascum sinuatum L. flowers (VSFs) is due to their potential applications across multiple sectors, including the pharmaceutical, cosmeceutical, and food industries, particularly in the development of functional foods and nutraceuticals. The phytochemical composition of VSFs is predominantly characterized by phenolic compounds, iridoid glycosides, and alkaloids such as plantagonine. This bioactive alkaloid identified in VSFs has been reported to have anticancer, antioxidant, and antimicrobial activities. In the literature, no studies have addressed the optimization of plantagonine extraction from VSFs. Thus, this study aimed to maximize the extraction of plantagonine from VSFs through microwave-assisted extraction (MAE) and pressurized liquid extraction (PLE). For MAE, 2g of dried VSF powder was placed in a closed vessel and dissolved in 40 mL of a solvent at various concentrations (S) and temperatures (T) and for various periods of time (t). Regarding PLE, the procedure involved placing 1g of dried VSFs in a 66 mL extraction cell containing diatomaceous earth under 10 bar pressure and applying various S, T and t conditions. Efficient recovery, meaning the maximization of the yield using the lowest possible inputs, was determined using the Response Surface Methodology (RSM) with three independent variables (t/X₁, T/X₂, and the proportion of ethanol in the hydroethanolic solution, S/X₃), employing a five-level circumscribed central composite design (CCCD) comprising 28 experimental runs. The plantagonine content in the VSF extracts obtained under the operational conditions for each point of the experimental design was evaluated through HPLC-ESI-QqQ-MS/MS. The results revealed that under the optimal extraction conditions, the yield obtained using PLE (2312.86 ± 39.69 µg/g extract; 25 min, 200°C, 59% ethanol) was 1.7-fold higher than that obtained using MAE (1356.20±32.10 µg/g extract; 25 min, 180°C, 55% ethanol). Therefore, this study proposes, for the first time, the optimal extraction conditions for maximizing the recovery of plantagonine from VSFs using two green extraction techniques, demonstrates the superiority of PLE, and provides a quantitative framework for further industrial extraction and application in the food, cosmeceutical, and pharmaceutical industries.

Corn silk (CS) (Stigma maydis) has been used for centuries in traditional medicine (TM) worldwide to treat various disorders owing to its antioxidant, antidiabetic, diuretic, antimicrobial, and anti-inflammatory properties. The health benefits mentioned in traditional medicine and reliable sources originate from the chemical makeup of corn silk, which includes total phenols, polysaccharides, protein, fiber, alkaloids, minerals, and vitamins. Large quantities of CS, a major by-product of corn processing, are often discarded, dissipating valuable bio-resources. By utilizing CS to develop functional and health-promoting value-added products, a part of agricultural waste would be redirected toward zero-waste production while conserving the environment and increasing the agro-economy. Our present study focused on investigating the bioactive compounds and antioxidant capacities of CS from five corn genotypes harvested at three maturity stages, or three different days of sillking (DAS) in order to single out the most promising raw material that could be used to develop dietary supplements. The prevailing bioactive compounds were total phenols, which ranged from 2498.09 GAE/g (genotype ZP 6119k, 15 DAS) to 13580.62 GAE/g (genotype ZP 5550, 15 DAS). Assessment of antioxidant capacity using different methods will show which corn silk genotypes are most suitable for health development. The antioxidant capacities determined after ABTS⁺ scavenging activity ranged from 41.82 mmol Trolox/kg to 83.48 mmol Trolox/kg in CS extracts of two genotypes, ZP 6119k (15 DAS) and ZP 6263 (21 DAS), respectively. The results obtained in this study could be of exceptional importance for the maize breeding programs and selection of potentially most suitable hybrids for functional food and dietary supplement production.

The global demand for plant-based dairy milk alternatives (PBDMAs) has risen steadily in recent years, driven by increasing consumer interest in health, sustainability, and dietary diversity. Simultaneously, the smoothie market has experienced consistent growth, with fruit-based smoothies incorporating PBDMAs becoming more prominent in contemporary diets. Despite their popularity, limited comparative research exists on the physicochemical characteristics of smoothies made with PBDMAs versus traditional dairy milk. This study investigates the physicochemical properties of strawberry smoothies prepared with dairy milk and PBDMAs. Thirteen formulations were analyzed: three with dairy milk (full cream, skim, and light), nine with different PBDMAs, and one with water as a dairy milk-free and PBDMA-free control. The pH values ranged from 3.81 (coconut milk smoothie) to 5.08 (soy milk smoothie), while total soluble solids (TSS) varied from 3.13 °Bx (control) to 10.13 °Bx (oat milk smoothie). Titratable acidity (TA) was close to zero, indicating an almost complete neutralisation of the characteristic acidity of strawberries. Colour analysis revealed a general decrease in lightness (L*) and an increase in redness (a*) across all samples. These findings highlight significant variations in the physicochemical properties of strawberry smoothies depending on whether dairy milk or PBDMAs were used as the base. Further research is needed to assess how these differences influence nutritional quality and consumer acceptance.

[Introduction] Honey's effects on maintaining beautiful skin, hematopoiesis, etc. have been reported. Meanwhile, the unique HP-SPR (high-precision surface Plasmon resonance) method enables the transcription measurement of polarization (waves) and the activity measurement of intracellular mitochondria. In this study, the effects of honey contact were examined from the transcription of polarization, and the effects of oral intake were examined from the activation of mitochondria in skin fibroblast (dermis) cells. The results were compared between hyperpolarized honey and a commercial product.

[Methods] Blends of monofloral honey from Yamaguchi, Japan, with maximum polarization transfer were used as hyperpolarized samples, and commercial products served as controls. PBS was placed inside the glass ring, an insulator on the HP-SPR sensor, and a 120-fold diluted sample was placed outside the ring and polarization (wave) transfer was measured. For oral intake, honey was digested in the stomach and via a duodenal step with enzymes, and samples with a molecular weight ≤ 10,000 were used. Mitochondrial polarization changes were measured by the HP-SPR-3D assay using the human skin fibroblast HFb16d.

[Results] Hyperpolarized honey showed a very high polarization transfer capacity, about 13 times higher than the commercial product at 400 seconds. Water in the vicinity contacted by hyperpolarized honey is polarized through insulators, and is expected to activate the throat, intestines, skin, hair, scalp, and other parts of the body. On the other hand, oral intake of hyperpolarized honey showed a concentration-dependent polarization of mitochondria, reaching approximately 3.5 times higher at 1.0 mg/mL than at 0.5 mg/mL. In contrast, at 1.0 mg/mL of the commercial product, mitochondrial polarization was only about one-fifth that of hyperpolarized honey.

[Conclusions] Hyperpolarized honey is expected to prevent and improve wrinkles (anti-aging) by activating skin fibroblasts (dermis) cells, and may also contribute to the activation of various types of cells through contact and ingestion.

Whole grain wheat flour is valued for its nutritional and antioxidant properties due to its high levels of phenolic compounds. However, its high bran content can negatively impact dough handling, bread texture, and colour, posing a challenge for developing healthier baked products with desirable sensory qualities. While sieving is commonly used to separate bran from endosperm, little is known about how controlled flour fractionation at specific particle sizes (250 µm and 500 µm) affects the distribution of bioactive compounds and flour functionality. Hence, this study investigated the impact of flour fractionation by sieving (250 µm and 500 µm) on colour, water absorption capacity, total phenolic content (TPC), total flavonoid content (TFC), antioxidant activity (2,2-diphenyl-1-picrylhydrazyl [DPPH] and 2,2'-azino-bis (3-ethylbenzothiazoline-6-sulfonic acid) [ABTS]), and selected phenolic compound composition. Whole white wheat flour (WWF) and whole brown wheat flour (BWF) were included for comparison. BWF showed the highest water absorption (0.84 mL/g), while fractionated flours (0.65–0.69 mL/g), particularly the 250 µm fraction, demonstrated improved lightness and higher TPC (approx. 3.2 mg QE/g) compared to BWF (1.39 mg QE/g) and WWF (1.97 mg QE/g). Although BWF retained higher antioxidant activity, fractionation significantly influenced phenolic compound profiles. Trans-ferulic acid was the predominant compound, and quercetin levels remained consistent across all samples. Principal component analysis (PCA) of TPC, TFC, phenolic acids, and antioxidant activities (ABTS and DPPH) explained over 75% of the total variance in the first two components, revealing distinct clustering by flour type and fraction. These findings highlight the potential of targeted flour fractionation as a novel strategy to enhance both the functional and nutritional properties of whole grain flours, offering a balanced solution for improving whole grain bread making without the need for full bran removal. Future studies should apply the flour in baked goods, including bread, muffins, and cakes.

The global shift toward plant-based proteins is driven by the need for sustainable food systems, improved public health, and enhanced food security. However, traditional extraction methods, such as alkaline extraction–isoelectric precipitation (AE–IEP), often result in protein denaturation, limited functionality, and increased environmental impact. These limitations have prompted efforts to develop more efficient and sustainable technologies.

This study systematically examines peer-reviewed studies comparing conventional techniques (AE–IEP and salt and solvent extraction) with innovative approaches such as ultrasound-assisted extraction (UAE), pulsed electric field (PEF), microwave-assisted extraction (MAE), ionic liquid extraction, and membrane-based purification. Articles were searched in Web of Science and Scopus using the keywords “Plant Protein Extraction” and “Purification.” From 3,761 initial records, duplicates, non-English texts, and studies without methodological detail were excluded. After screening and eligibility checks, 124 full-text articles were assessed, and 72 studies were finally included in the synthesis.

The synthesis of the current literature shows that modern approaches are being applied across a wide range of crops and biological sources, from cereals and legumes to oilseeds, algae, and underutilized plants. Emerging methods consistently improve extraction efficiency, enhance solubility and emulsification properties, and reduce contaminants such as phenolics, anti-nutritional compounds, and toxins. Research also highlights the integration of nanotechnology, enzymatic modification, and microbial or biopolymer-based approaches, which expand functional and therapeutic applications. Studies on protein–polyphenol complexes, bioactive peptides, extracellular vesicles, and enzyme-assisted detoxification further demonstrate how extraction and purification are increasingly linked to health-related benefits, not just yield and purity.

Overall, modern technologies provide substantial improvements in the quality and functionality of plant protein isolates. Yet, challenges remain in balancing yield, purity, and cost, as well as scaling laboratory advances into industrial processes. These advances, when adapted to different plant sources and market needs, hold strong potential for building resilient protein systems that meet both nutritional demands and sustainability goals.