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- Fish processing industry residues: a review of valuable products extraction and characterization methodsPublication . Ideia, Pedro; Pinto, Joana; Ferreira, Rui; Figueiredo, Leandro; Spínola, Vítor; Castilho, Paula C.Fish processing industry has experienced significant growth, playing an important role in the world economy. The increased exploration of marine resources contributes to the generation of considerable amounts of biowaste, which ends up as discards. In the face of the resultant disposal and environmental problems, many efforts have been made to deal with the fishery waste in more efficient ways. Nowadays, these by-products are regarded as important sources of high added value compounds, such as hydroxyapatite, collagen, gelatin, lipids, enzymes, hydrolysates and bioactive peptides, with great potential for human health applications. The present paper aims to review the current methods of extraction and characterization of added value products from fish by-products, as well as their actual and potential applications.
- Hypoglycemic, anti-glycation and antioxidant in vitro properties of two Vaccinium species from Macaronesia: A relation to their phenolic compositionPublication . Spínola, Vítor; Pinto, Joana; Castilho, Paula C.Vaccinium cylindraceaeum (Azores blueberry) and V. padifolium (Madeira blueberry) are two Portuguese endemic species, which phytochemical composition and beneficial properties lack scientific validation. In this work, their potential in vitro inhibitory effects against key enzymes linked to type 2 diabetes and obesity and glycation of proteins were evaluated. Anthocyanins were predominant in fruits, while hydroxycinnamic acids (in particular 5-O-caffeoylquinic acid) and flavonoids were the main compounds in leaves. Vaccinium extracts showed effective inhibitions of glucosidases and glycation of proteins and mild activity towards α-amylase and pancreatic lipase (by comparison with reference compounds). Phenolic composition was strongly correlated with reported bioactivities. In fact, 5-O-caffeoylquinic can be considered one of the main hypoglycemic and anti-glycation agents of analysed extracts. Consumption of both Vaccinium species should be encouraged since berries could offer a dietary option in the prevention and control of diabetes and obesity, while leaves are good candidates for development of nutraceuticals.
- Release of health-related compounds during in vitro gastro-intestinal digestion of okara and okara fermented with Lactobacillus plantarumPublication . Quintana, Gabriel; Spínola, Vítor; Martins, Gonçalo N.; Gerbino, Esteban; Gómez-Zavaglia, Andrea; Castilho, Paula C.Okara is a highly perishable by-product remaining after filtration of the smashed soybeans seeds in the production of soymilk. Due to its nutritional value, different approaches have been developed to use it as functional ingredient. Fermentation of okara appears as an interesting strategy to preclude spoilage, providing a more stable matrix to be incorporated in the formulation of functional foods. Okara has antioxidant compounds but the effect of fermentation, and their bioaccessibility still need to be investigated. To achieve this aim, the phenolic compounds (as determined by TPC and TFC assays) and the antioxidant properties (as determined by ABTS ·+, DPPH · , O2·- assays) of okara and okara fermented with Lactobacillus plantarum CIDCA 83114 were assessed both before and after exposure to simulated gastro-intestinal conditions. Before digestion, okara showed higher values of TPC and TFC than the fermented counterpart. Although a decrease of TPC and TFC was observed after exposing okara to gastric conditions, no significant differences between okara and fermented okara were detected. No further decrease of TPC were observed in intestinal conditions. Okara showed higher antioxidant activity than fermented okara. There was a considerable decrease in the antioxidant activity for both samples when exposed to gastric and intestinal conditions. A good correlation between TFC and antioxidant activities was detected, suggesting that flavonoids play an important role as antioxidants. As a whole, this work provides a solid support for the stability of phytochemicals along the digestive process of both okara and fermented okara.
- Acid-modified clays as green catalysts for the hydrolysis of hemicellulosic oligosaccharidesPublication . Vilcocq, Léa; Spínola, Vítor; Moniz, Patrícia; Duarte, Luís C.; Carvalheiro, Florbela; Fernandes, César; Castilho, PaulaThe hydrolysis of hemicellulosic oligosaccharides (OS) was investigated using acid-activated clays (prepared from natural Porto Santo montmorillonite clay) as catalysts. Acid activation was performed in HCl solution or with aluminium exchange. The clay catalysts were characterized by XRD, N2 adsorption isotherms, CEC, FTIR, titration of acid sites in water and adsorption of sugars and disaccharides. They were tested for the hydrolysis of a model compound, maltose, and of OS-rich liquor from rice straw fractionation. The HCl-activated clays were the most efficient catalysts for maltose hydrolysis. It was demonstrated that the hydrolysis of OS into monomer sugars over a clay catalyst is technically feasible and that this reaction leads to the selective removal of glucose, arabinose and acetic acid side groups from the OS structure, thus yielding simpler xylo-oligosaccharide chains. Furthermore, no significant conversion of monomer sugars into furans was observed.
- An improved and fast UHPLC-PDA methodology for determination of L-ascorbic and dehydroascorbic acids in fruits and vegetables. Evaluation of degradation rate during storagePublication . Spínola, Vítor; Mendes, Berta; Câmara, José S.; Castilho, Paula C.This study provides a versatile validated method to determine the total vitamin C content, as the sum of the contents of L-ascorbic acid (L-AA) and dehydroascorbic acid (DHAA), in several fruits and vegetables and its degradability with storage time. Seven horticultural crops from two different origins were analyzed using an ultra-high-performance liquid chromatographic-photodiode array (UHPLC-PDA) system, equipped with a new trifunctional high strength silica (100% silica particle) analytical column (100 mm × 2.1 mm, 1.7 μm particle size) using 0.1% (v/v) formic acid as mobile phase, in isocratic mode. This new stationary phase, specially designed for polar compounds, overcomes the problems normally encountered in HPLC and is suitable for the analysis of large batches of samples without L-AA degradation. In addition, it proves to be an excellent alternative to conventional C18 columns for the determination of L-AA in fruits and vegetables. The method was fully validated in terms of linearity, detection (LOD) and quantification (LOQ) limits, accuracy, and inter/intra-day precision. Validation experiments revealed very good recovery rate of 96.6 ± 4.4% for L-AA and 103.1 ± 4.8 % for total vitamin C, good linearity with r(2)-values >0.999 within the established concentration range, excellent repeatability (0.5%), and reproducibility (1.6%) values. The LOD of the method was 22 ng/mL whereas the LOQ was 67 ng/mL. It was possible to demonstrate that L-AA and DHAA concentrations in the different horticulture products varied oppositely with time of storage not always affecting the total amount of vitamin C during shelf-life. Locally produced fruits have higher concentrations of vitamin C, compared with imported ones, but vegetables showed the opposite trend. Moreover, this UHPLC-PDA methodology proves to be an improved, simple, and fast approach for determining the total content of vitamin C in various food commodities, with high sensitivity, selectivity, and resolving power within 3 min of run analysis.
- Ulex europaeus: from noxious weed to source of valuable isoflavones and flavanonesPublication . Spínola, Vítor; Llorent-Martínez, Eulogio J.; Gouveia-Figueira, Sandra; Castilho, Paula C.The screening and quantification of the main phenolic compounds in leaves and flowers of Ulex europaeus (gorse) was carried out by high-performance liquid chromatography with electrospray ionization mass spectrometric detection (HPLC-ESI–MSn) after ultrasound-assisted extraction with methanol. About 98% of compounds corresponded to flavonoids, distributed as flavonols, flavones, isoflavones and flavanones. Flavonols were mainly quercetin glucosides; most of the found flavones were apigenin derivatives and the isoflavone group was dominated by glycitin. The flavanone group was composed mainly of liquiritigenin derivatives, substances usually found in liquorice (Glycyrrhiza ssp) and associated with high pharmacological relevance; in Ulex they represent about 25% of total polyphenols content. Phenolic acids and saponins were also detected, as minor components. In vitro antioxidant activity (nitric oxide, superoxide assays, ABTS and DPPH assays) of leaves and flowers, and their inhibitory effects towards digestive enzymes related to carbohydrate metabolism (-glucosidase and -amylase) were also studied.
- Polyphenols of Myrica faya inhibit key enzymes linked to type II diabetes and obesity and formation of advanced glycation end-products (in vitro): Potential role in the prevention of diabetic complicationsPublication . Spínola, Vítor; Llorent-Martínez, Eulogio J.; Castilho, Paula C.Myrica faya Aiton (fire tree, faya) is an underused species with a diverse flavonoid composition (anthocyanins, flavonols, ellagitannins) which can promote positive effects on human health. M. faya has been reported to possess high antioxidant activities, but its potential in the prevention of type II diabetes has not been evaluated so far. In the present study, eight M. faya samples from different areas of Madeira and Azores archipelagos (Portugal) were collected to determine their phytochemical profile and then tested for their in vitro anti-diabetic and antioxidant activities. The analyzed extracts showed strong inhibitory activities towards α -glucosidase, aldose reductase and glycation of bovine serum albumin (BSA) and moderate effects towards α-amylase and lipase (by comparison with reference compounds). Cyanidin-3-O-glucoside and ellagitannins were the main bioactive agents involved in the anti-diabetic effects of M. faya. Such results may provide important scientific evidence for further utilization of M. faya as dietary or nutraceutical products for the prevention and/or control of hyperglycaemia-associated complications.
- Evaluation of Asteraceae herbal extracts in the management of diabetes and obesity. Contribution of caffeoylquinic acids on the inhibition of digestive enzymes activity and formation of advanced glycation end-products (in vitro)Publication . Spínola, Vítor; Castilho, Paula C.The study was performed to assess, for the first time, the in vitro anti-diabetic potential of ten Asteraceae plant extracts to inhibit the activity of digestive enzymes (α-amylase, α-, β-glucosidases and lipase) responsible for hydrolysis/digestion of sugar and lipids. Prevention of advanced glycation end-products (AGEs) formation was evaluated in bovine serum albumin/ribose glycation reaction model. The phytochemical profiles and caffeoylquinic acids (CQAs) contents were determined for the methanolic extract of each plant. Analyzed plant extracts exhibited significant inhibitory activity against key digestive enzymes linked to type II diabetes and obesity. A strong inhibition was observed for glucosidases and mild activity towards amylase and lipase (compared to reference compounds). Moreover, some extracts exhibited potent ability to prevent formation of AGEs, implicated in some diabetic complications. Caffeoylquinic acids were dominant in all plant extracts and findings demonstrate that these compounds are the most relevant hypoglycemic and anti-glycation agents. From the obtained results, Argyranthemum pinnatifidum, Helichrysum melaleucum, and Phagnalon lowei are good candidates for further development of phyto-pharmaceutical preparations as complementary therapy for diabetes and obesity control.
- Effect of time and temperature on vitamin C stability in horticultural extracts. UHPLC-PDA vs iodometric titration as analytical methodsPublication . Spínola, Vítor; Mendes, Berta; Câmara, José S.; Castilho, Paula C.Several fruits and vegetables from Madeira Island (Portugal) were evaluated by two analytical methods for their total vitamin C content (l-ascorbic acid, l-AA and dehydroascorbic acid, DHAA). DHAA was determined indirectly with DL-1,4-dithiotreitol (DTT) applied as a pre-column reductant. Ultra high performance liquid chromatography coupled to photodiode array (UHPLC-PDA) determinations were compared with l-AA content obtained by a classic iodometric titration method. The stability of vitamin C in horticultural extracts stored at different temperatures was also investigated. Red peppers represented the better source of vitamin C followed by green peppers and papayas. Passion fruits and cherimoyas were the analyzed foodstuffs with lowest vitamin C content. Both analytical methods were suitable for l-AA analysis in various food commodities, the UHPLC-PDA technique being preferred due to its advantages of selectivity, speed and accuracy. The degradation study showed that horticultural extracts were stable at least 24 h at 4 °C and during 4 weeks when stored at −80 °C.
- Identification and quantification of phenolic compounds of selected fruits from Madeira Island by HPLC-DAD–ESI-MSn and screening for their antioxidant activityPublication . Spínola, Vítor; Pinto, Joana; Castilho, Paula C.Five fruits species commonly cultivated and consumed in Madeira Island (Portugal) were investigated for their phenolic profile by means of reversed phase high-performance liquid chromatography coupled to diode array detection and electrospray ionisation mass spectrometry (HPLC-DAD-ESI/MS(n)) and antioxidant potential. A large number of compounds were characterised, flavonoids and phenolic acids being the major components found in target samples, 39 compounds (flavonoids, phenolic acids, terpenoids, cyanogenic glycosides and organic acids) were identified in cherimoyas, lemons, papayas, passion-fruits and strawberries for the first time. Furthermore, all samples were systematically analysed for their total phenolic and flavonoid contents along with two radical scavenging methods (ABTS and ORAC) for antioxidant activity measurement. Target fruits presented high phenolic contents which is responsible for most of the antioxidant activity against radical reactive species (R(2)>0.80). Quantitative data showed that anthocyanins, in particular pelargonidin-3-O-hexoside (>300 mg/100 mL), present only in strawberries were the compounds in largest amounts but are the ones which contribute less to the antioxidant activity.
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