Fernando Benavente

β-Casein A1 and A2 (β-CN-A1 and β-CN-A2) are the two predominant β-CN proteoforms in bovine milk. β-CN-A1 has been associated with a greater propensity to release opioid peptides, such as β-casomorphin-7 (β-CM-7) and β-casomorphin-5 (β-CM-5), during gastrointestinal (GI) digestion, which may have adverse biological effects. This has stimulated growing interest in milk from cows carrying the β-CN A2A2 genotype (A2 milk), which requires reliable characterization methods. In this work, we developed a rapid, selective, and sensitive capillary electrophoresis–mass spectrometry (CE-MS) method for the accurate identification and quantification of β-CM-7 and β-CM-5 in milk hydrolysates from in vitro GI digestion of bovine milk. The method showed good linearity (R² > 0.99, over 0.5–100 mg/L for β-CM-7 and 0.25–100 mg/L for β-CM-5), limits of detection (0.25 and 0.10 mg/L), and repeatability (<0.2% for times and <1.4% for areas), and tandem mass spectrometry (MS/MS) allowed confirmation. The method was applied to A1A1 and A2A2 milk digested using the standardized INFOGEST protocol, followed by solid-phase extraction. β-CM-7 was detected and quantified only in A1A1 digests (0.98 mg/L), whereas β-CM-5 was not detected (<0.10 mg/L). These results indicate a differential release of β-CMs from A1 and A2 milk and support the method’s suitability for β-CM profiling, which may help assess A2 milk quality control and β-CM health impact.

Skin aging is a complex and irreversible natural process impacted by genetics, lifestyle, and environmental variables, leading to wrinkles, loss of elasticity, and uneven skin tone. There is a growing demand for natural skincare products, which are perceived as safer, more sustainable, and effective in combating the signs of aging. Clerodendrum infortunatum ( C. infortunatum ) Linn. (Family Lamiaceae) has traditional medicinal uses, including hepatoprotection, antimicrobial, and vermifuge activity, as well as benefits in alleviating inflammation, arthritis, and diabetes. This study aims to investigate the metabolites present in different solvent-extracted fractions of the aerial parts of C. infortunatum through liquid chromatography-tandem mass spectrometry (LC-MS/MS) profiling, and to evaluate their anti-aging potential using in vitro anti-collagenase and anti-elastase assays. The correlation between the identified metabolites and bioactivity was investigated using a partial least squares (PLS) chemometric approach. To confirm these findings, molecular docking studies were performed to assess the inhibitory potential of the metabolites most strongly correlated with bioactivity against elastase and collagenase target enzymes. This study is the first to correlate the secondary metabolites of C. infortunatum solvent-extracted fractions with their newly discovered anti-aging properties, representing a significant contribution to the field. Additionally, it presents the first molecular docking analysis of salsaside A, jionoside C, and 6′-caffeoyl-12-glucosyloxy-jasmonic acid against collagenase and elastase enzymes, revealing significant binding affinities and promising inhibitory potential. These findings underscore the potential of these metabolites as novel anti-aging compounds through their strong interactions with key target enzymes.

In the present study, we investigated the potential biological effects of proteins and peptides extracted from four different commercial quinoa grain varieties: Black (B), Red (R), White (W), and Royal White (RO), using Caco‐2 cells as a proliferation model. Concentration–response curves were constructed to evaluate cytotoxicity and antiproliferative effects. Interestingly, peptides exhibited higher cytotoxicity than proteins, particularly in R and RO quinoa varieties. Based on these findings, we used a data mining approach to identify and compare the cytotoxic proteins in the four quinoa varieties. Using their relative abundance, we successfully classified R and RO quinoa as a cohesive group using classification models. To gain deeper insights into the biological effects on cells, we comprehensively analyzed the differential expression of apoptosis biomarkers using liquid chromatography‐tandem mass spectrometry (LC‐MS/MS) targeted proteomics. Finally, we correlated the apoptosis profile with the relative abundance of cytotoxic proteins. All these studies were supported by the application of multivariate data analysis. The results demonstrated the potential anticancer of quinoa grain proteins and peptides and provided the basis for more focused mechanistic studies aimed at developing functional foods and nutraceuticals.

Jojoba (Simmondsia chinensis) seeds are broadly used worldwide for their valuable oil. The chemical composition of the other parts of the jojoba shrub remains largely unexplored. In this work, the metabolite profiles of aqueous and methanolic extracts from different jojoba plant organs were analyzed by liquid chromatography-quadrupole time-of-flight tandem mass spectrometry (LC-QTOF-MS/MS) combined with molecular networking (MN), resulting in the assignment of 104 metabolites from different classes. All methanolic extracts evinced significant antibacterial activity against the examined bacterial strains. Subminimum inhibitory concentrations (sub-MICs) of the hull and leaf extracts successfully inhibited biofilm formation of Staphylococcus aureus with inhibition rates above 90%. The seed extract at 0.5 MIC effectively eradicated S. aureus and Enterococcus faecalis biofilms (>80%). Multivariate data analysis revealed the positively correlated biomarkers with bioactivity, primarily flavonoids, phenolic compounds, and simmondsins. These results present a ground for future research to valorize jojoba seed-derived products and byproducts from other organs.

Quinoa is widely recognized for its exceptional nutritional properties, particularly its complete protein content. This study, for the first time, investigates the effects of processing methods (boiling and extrusion) and farming conditions (conventional and organic) on the proteomic profile. Following a label-free shotgun proteomics approach, a total of 1796 proteins were identified and quantified across all quinoa samples. Regarding processing, both boiling and extrusion produced protein extracts with lower total protein content, with the number of identified proteins decreasing from 1695 in raw quinoa to 957 in processed quinoa. Boiling led to a reduction in protein diversity and expression, while extrusion, which involves high temperatures and pressures, specifically decreased the abundance of high molecular mass proteins. Concerning cultivation practices, organic farming was associated with a broader protein diversity, especially proteins related to translation (28 vs 5%), while conventional farming showed a higher abundance of catalytic and enzymatic proteins (67 vs 46%). These findings highlight the distinct proteomic changes induced by different processing methods and farming conditions, offering valuable insights to manage quinoa's nutritional, bioactive, and functional properties across various production practices.