Harinas de leguminosas deshidratadas: caracterización nutricional y valoración de sus propiedades tecnofuncionales

Tesis doctoral inédita. Universidad Autónoma de Madrid, Facultad de Ciencias, Departamento de Química Agrícola. Fecha de lectura: 14-01-201

Biorefinery and Stepwise Strategies for Valorizing Coffee By-Products as Bioactive Food Ingredients and Nutraceuticals

Featured Application: This study presents a comprehensive and stepwise strategy for utilizing coffee by-products as a sustainable source to develop bioactive food ingredients and nutraceuticals. Coffee by-products can be transformed into versatile functional food ingredients and nutraceuticals by implementing green extraction techniques and employing a biorefinery approach. These products offer a wide range of benefits, including antioxidant, anti-inflammatory, and anti-obesity effects, as well as the potential to regulate energy metabolism and blood sugar levels and serve as adjunct therapies for conditions such as cardiovascular disease, diabetes, and neurodegenerative disorders. This research not only contributes to the creation of functional food products and supplements but also promotes sustainability and addresses the prevention and management of chronic diseases. Coffee production generates significant amounts of by-products, posing challenges for waste management in the industry. Recent research has revealed that coffee by-products are rich in bioactive compounds suitable to produce functional food ingredients and nutraceuticals. In this review, we explore biorefinery strategies for extracting and utilizing bioactive compounds from coffee by-products, including the production of bio-based chemicals and materials, as well as the extraction of phenolic compounds, antioxidants, and dietary fiber for food applications. We propose a stepwise approach for the development of functional food ingredients and nutraceuticals from coffee by-products, covering the identification of needs, comprehensive characterization, in vitro and in vivo research, unraveling the mechanism of action, food and nutraceutical formulation, sensory analysis, shelf-life stability, scale-up, randomized control trials, and biostatistics and bioinformatic integration. Additionally, we discuss the market potential, regulatory issues, and technological innovation surrounding the commercialization of coffee by-product-based products. Emphasizing the importance of regulatory compliance and sustainability in the coffee industry, this review highlights the potential of coffee by-products to be transformed from waste into valuable functional food ingredients and nutraceuticals, offering a promising avenue for waste reduction and promoting sustainability in the coffee industryThis research was funded by the COCARDIOLAC (RTI 2018-097504-B-I00) and ECOPULP (TED2021-129262A-I00) projects from the Spanish Ministry of Science and Innovation and the Excellence Line for University Teaching Staff within the Multiannual Agreement between the Community of Madrid and the UAM (2019–2023). M. Rebollo-Hernanz received funding from the FPU program of the Ministry of Universities for his predoctoral fellowship (FPU15/04238) and a grant for the requalification of the Spanish university system (CA1/RSUE/2021-00656)

Bioaccessibility of phenolic compounds from cocoa shell subjected to in vitro digestion and its antioxidant activity in intestinal and hepatic cells

The cocoa shell is a by-product generated by the cocoa processing industry that could be used as a nutraceutical owing to the significant amounts of bioactive compounds it contains. This work aimed to study the bioaccessibility of phenolic compounds present in the flour (CSF) and an aqueous extract (CSE) from cocoa shells through an in vitro simulated digestion and to assess their antioxidant capacity in vitro by using intestinal and hepatic cell culture models (IEC-6 and HepG2 cells). The bioaccessibility of phenolic compounds was determined using a simulated in vitro digestion model (INFOGEST). Total phenolic compounds (TPC) and antioxidant activity were measured using in vitro techniques. Reactive oxygen species (ROS) were evaluated in IEC-6 and HepG2 cells after t-BOOH stimulation. TPC present in CSE were more bioaccessible than phenolic compounds present in CSF. During digestion, the bioaccessibility of phenolic compounds from CSF fluctuated in the gastric (2.8 mg/g), intestinal (7.6 mg/g), and colonic (5.7 mg/g) phases. Similarly, for the phenolics of CSE, the bioaccessibility increased from 50.6 mg/g in the gastric phase to 53.4 mg/g in the intestinal phase and decreased in the colonic phase to 37.2 mg/g. The in vitro antioxidant capacity followed a similar behavior, increasing throughout the digestion in CSF (8.8- to 10.6-fold) and CSE (6.0- to 7.4-fold). Digested CSF and CSE were not cytotoxic for IEC-6 and HepG2 cells and protected their viability under oxidative stress conditions (93–100%). t-BOOH-induced ROS were prevented by CSF (72–88%) and CSE (81–94%) bioaccessible fractions in both intestinal and hepatic cells. In conclusion, cocoa shells are a source of potentially bioavailable antioxidant phenolic compounds that may protect cells from oxidative stress

Evaluation of parameters which influence voluntary ingestion of supplements in rats

Drug safety and efficacy studies frequently use oral gavage, but repetitive usage may cause problems. Administration through voluntary ingestion represents an opportunity for refinement. We aimed to develop a protocol for voluntary ingestion of gelatin-based supplements in rats, assessing the influence of age, sex, fasting (4 h), and additives (vanilla, VF; sucralose, S), and to test it in lactating dams. Three-week-old and 5-month-old Sprague-Dawley rats were placed individually in an empty cage containing a gelatin cube and trained daily (5 days/week), recording the day the whole cube was consumed (latency). Rats trained prior to gestation were offered a gelatin containing 250 mg/kg cocoa shell extract (CSE) during lactation. Rats that did not eat the cube after 8 training days were considered non-habituated, with a proportion similar in young males (7.1%), young females (11.1%), and adult females (10.3%), but significantly higher in adult males (39.3%). Excluding non-habituated rats, latency was 2–3 days, without differences between young and adult rats (p = 0.657) or between males and females (p = 0.189). VF or VF + S in the gelatin did not modify latency, while fasting significantly reduced it in females (p = 0.007) but not in males (p = 0.501). During lactation, trained females ate the CSE-gelatin within 1–5 min without litter problems. Conclusions: Acceptance of a gelatin-based supplement is negatively influenced by male sex, facilitated by fasting, and not modified by additives. Training is remembered after 2 months and does not interfere with lactation. Gelatin-based voluntary ingestion is suitable to administer drugs that need to pass through the digestive system, ensuring adequate dosage, and is important to detect non-habituated rats prior to the study. The current protocol may be implemented by training the rats in their own cageRTI 2018-097504-B-I0

Understanding the gastrointestinal behavior of the coffee pulp phenolic compounds under simulated conditions

Numerous residues, such as the coffee pulp, are generated throughout coffee processing. This by-product is a source of antioxidant phytochemicals, including phenolic compounds and caffeine. However, the antioxidant properties of the phenolic compounds from the coffee pulp are physiologically limited to their bioaccessibility, bioavailability, and biotransformation occurring during gastrointestinal digestion. Hence, this study explored the phenolic and caffeine profile in the coffee pulp flour (CPF) and extract (CPE), their intestinal bioaccessibility through in vitro digestion, and their potential bioavailability and colonic metabolism using in silico models. The CPE exhibited a higher concentration of phenolic compounds than the CPF, mainly phenolic acids (protocatechuic, chlorogenic, and gallic acids), followed by flavonoids, particularly quercetin derivatives. Caffeine was found in higher concentrations than phenolic compounds. The antioxidant capacity was increased throughout the digestive process. The coffee pulp matrix influenced phytochemicals’ behavior during gastrointestinal digestion. Whereas individual phenolic compounds generally decreased during digestion, caffeine remained stable. Then, phenolic acids and caffeine were highly bioaccessible, while flavonoids were mainly degraded. As a result, caffeine and protocatechuic acid were the main compounds absorbed in the intestine after digestion. Non-absorbed phenolic compounds might undergo colonic biotransformation yielding small and potentially more adsorbable phenolic metabolites. These results contribute to establishing the coffee pulp as an antioxidant food ingredient since it contains bioaccessible and potentially bioavailable phytochemicals with potential health-promoting propertie

Exploring the potential of phenolic compounds from the coffee pulp in preventing cellular oxidative stress after in vitro digestion

The coffee pulp, a by-product of the coffee industry, contains a high concentration of phenolic compounds and caffeine. Simulated gastrointestinal digestion may influence these active compounds’ bioaccessibility, bioavailability, and bioactivity. Understanding the impact of the digestive metabolism on the coffee pulp's phenolic composition and its effect on cellular oxidative stress biomarkers is essential. In this study, we evaluated the influence of in vitro gastrointestinal digestion of the coffee pulp flour (CPF) and extract (CPE) on their phenolic profile, radical scavenging capacity, cellular antioxidant activity, and cytoprotective properties in intestinal epithelial (IEC-6) and hepatic (HepG2) cells. The CPF and the CPE contained a high amount of caffeine and phenolic compounds, predominantly phenolic acids (3′,4′-dihydroxycinnamoylquinic and 3,4-dihydroxybenzoic acids) and flavonoids (3,3′,4′,5,7-pentahydroxyflavone derivatives). Simulated digestion resulted in increased antioxidant capacity, and both the CPF and the CPE demonstrated free radical scavenging abilities even after in vitro digestion. The CPF and the CPE did not induce cytotoxicity in intestinal and hepatic cells, and both matrices exhibited the ability to scavenge intracellular reactive oxygen species. The coffee pulp treatments prevented the decrease of glutathione, thiol groups, and superoxide dismutase and catalase enzymatic activities evoked by tert-butyl hydroperoxide elicitation in IEC-6 and HepG2 cells. Our findings suggest that the coffee pulp could be used as a potent food ingredient for preventing cellular oxidative stress due to its high content of antioxidant compoundsThis research was funded by the COCARDIOLAC project from the Spanish Ministry of Science and Innovation (RTI 2018-097504-B-I00) and the Excellence Line for University Teaching Staff within the Multiannual Agreement between the Community of Madrid and the UAM (2019-2023). M. Rebollo-Hernanz received funding from the FPU program of the Ministry of Universities for his predoctoral fellowship (FPU15/04238) and his grant for the requalification of the Spanish university system (CA1/RSUE/2021-00656

Improvement of the Seminal Characteristics in Rams Using Agri-Food By-Products Rich in Phytomelatonin

The aim of this study was to evaluate the effect of a phytomelatonin-rich diet, including by-products from the food industry, on ram sperm quality and seminal plasma composition. Melatonin content in several by-products before and after in vitro ruminal and abomasal digestion was determined by HPLC-ESI-MS/MS. Finally, 20% of a mix of grape pulp with pomegranate and tomato pomaces was included in the rams’ diet, constituting the phytomelatonin-rich diet. Feeding the rams with this diet resulted in an increase in seminal plasma melatonin levels compared with the control group (commercial diet) in the third month of the study. In addition, percentages higher than those in the control group of morphologically normal viable spermatozoa with a low content of reactive oxygen species were observed from the second month onwards. However, the antioxidant effect does not seem to be exerted through the modulation of the antioxidant enzymes since the analysis of the activities of catalase, glutathione reductase and glutathione peroxidase in seminal plasma revealed no significant differences between the two experimental groups. In conclusion, this study reveals, for the first time, that a phytomelatonin-rich diet can improve seminal characteristics in ram

Radical scavenging and cellular antioxidant activity of the cocoa shell phenolic compounds after simulated digestion

The cocoa industry generates a considerable quantity of cocoa shell, a by-product with high levels of methylxanthines and phenolic compounds. Nevertheless, the digestion process can extensively modify these compounds’ bioaccessibility, bioavailability, and bioactivity as a consequence of their transformation. Hence, this work’s objective was to assess the influence of simulated gastrointestinal digestion on the concentration of phenolic compounds found in the cocoa shell flour (CSF) and the cocoa shell extract (CSE), as well as to investigate their radical scavenging capacity and antioxidant activity in both intestinal epithelial (IEC-6) and hepatic (HepG2) cells. The CSF and the CSE exhibited a high amount of methylxanthines (theobromine and caffeine) and phenolic compounds, mainly gallic acid and (+)-catechin, which persisted through the course of the simulated digestion. Gastrointestinal digestion increased the antioxidant capacity of the CSF and the CSE, which also displayed free radical scavenging capacity during the simulated digestion. Neither the CSF nor the CSE exhibited cytotoxicity in intestinal epithelial (IEC-6) or hepatic (HepG2) cells. Moreover, they effectively counteracted oxidative stress triggered by tert-butyl hydroperoxide (t-BHP) while preventing the decline of glutathione, thiol groups, superoxide dismutase, and catalase activities in both cell lines. Our study suggests that the cocoa shell may serve as a functional food ingredient for promoting health, owing to its rich concentration of antioxidant compounds that could support combating the cellular oxidative stress associated with chronic disease developmentThis research was funded by the COCARDIOLAC project from the Spanish Ministry of Science and Innovation (RTI 2018-097504-B-I00) and the Excellence Line for University Teaching Staff within the Multiannual Agreement between the Community of Madrid and the UAM (2019–2023). M. Rebollo-Hernanz received funding from the FPU program of the Ministry of Universities for his predoctoral fellowship (FPU15/04238) and a grant for the requalification of the Spanish university system (CA1/RSUE/2021-00656