Halophytes are salinity–sensitive plants that tolerate extremely high salt concentrations, being mainly found in saltmarshes and coastal areas worldwide, including Portugal [1]. Halophytes, such as Salicornia spp., are used by human due to commercial, ecological and gastronomic values [2]. Inside this genus, Salicornia ramosissima is of particular interest, being considered an alternative to salt [3]. The valorization of S. ramosissima may be a sustainable and environmental-friendly way to obtain extracts with interest for nutraceutical industry. Nevertheless, extraction is a key step to acquire bioactive compounds from plants, with emphasis on the extraction technique and solvent employed, which should maximize the extraction yield [13]. The aim of this study was to recover bioactive compounds from S. ramosissima by conventional extraction (CE) and Microwave-assisted extraction (MAE) techniques using water as extractor solvent.

In this study, the total phenolic and flavonoid contents (TPC and TFC, respectively), antioxidant/antiradical activities (through FRAP and ABTS assays) as well as the phenolic profile and the in-vitro effects on intestinal cells were screened. Briefly, CE was performed as a decoction preparation while MAE was executed at selected temperatures (72 – 94 ºC) during 1 to 5 min with constant medium stirring. After extractions, samples were filtrated and lyophilized for further assays.

The values of TPC ranged between 8.34 and 15.02 mg of gallic acid equivalents (GAE)/g for CE and MAE extracts, respectively. In same line, CE extract exhibited the highest TFC value (8.44 mg of catechin equivalents (CAE)/g). Regarding ABTS and FRAP assays, the MAE extract showed the highest values (60.61 μmol FSE/g dw and 16.06 % inhibition for FRAP and ABTS assays, respectively). On the other hand, the total amount of phenolic compounds identified and quantified in both extracts were similar, being the phenolic acids and flavonols the principal constituents. For both extracts, myricetin was the compound present in highest amounts (0.4250 and 0.4655 mg myricetin/g dw for CE and MAE extracts, respectively) and gallic acid was the major phenolic acid present in extracts (0.2105 and 0.1553 mg gallic acid/g dw for CE and MAE extracts, respectively). The cell effects demonstrated that both extracts did not lead to a decrease of HT29-MTX viability. In Caco-2 cell line, only the highest concentration of MAE (1000 µg/mL) led to a decrease of viability (86.55%). In this sense, S. ramosissima extracted by CE or MAE can be classified as non-toxic and rich in bioactive compounds. However further studies, such as in-vitro intestinal permeation assays and in-vivo studies, are needed to underline this potential.

References:

[1] Flowers, T.J.; Colmer, T.D. Salinity Tolerance in Halophytes. New Phytol. 2008, 179, 945–963.

[2] Lima, A.R.; Castañeda-Loaiza, V.; Salazar, M.; Nunes, C.; Quintas, C.; Gama, F.; Pestana, M.; Correia, P.J.; Santos, T.; Varela, J.; et al. Influence of Cultivation Salinity in the Nutritional Composition, Antioxidant Capacity and Microbial Quality of Salicornia Ramosissima Commercially Produced in Soilless Systems. Food Chem. 2020, 333, 127525.

[3] Ferreira, D.; Pinto, D.C.G.A.; Silva, H.; Girol, A.P.; de Lourdes Pereira, M. Salicornia ramosissima J. Woods Seeds Affected the Normal Regenerative Function on Carbon Tetrachloride-induced Liver and Kidney Injury. Biomed. Pharmacother. 2018, 107, 283–291.

[4] Panja, P. Green Extraction Methods of Food Polyphenols from Vegetable Materials. Curr. Opin. Food Sci. 2018, 23, 173–182.

Water soluble synthetic food colorants are widely used in the production of beverages for improving their appearance, being highly appreciated by producers especially due to their stability, compatibility with the food matrix, tinctorial power and price. However, previous studies on the toxicity of these food additives highlighted several worrying health-related issues on consumers: an excessive consumption of certain synthetic food colorants can cause allergic reactions, attention deficit hyperactivity disorder (ADHD) in sensitive children, or even cancer. Because soft drinks are highly-consumed beverages amongst all age groups, especially teenagers and children, they can be considered as a health risk for consumers. The aim of the present work is provide data on the content of eight synthetic food colorants from several soft drinks available on the Romanian market: Allura Red (E 129), Azorubine (E 122), Brilliant Blue (E 133), Patent Blue (E 131), Ponceau 4R (E 124), Ponceau 6R (E 126), Tartrazine (E 102) and Sunset Yellow (E 110). Soft drinks were purchased from hypermarkets; representative samples were subjected to solid phase extraction (carbonated soft drinks were degassed on an ultrasonic bath before extraction), then filtered through 0.45 μm membrane filters. Analytical determinations were performed by high performance liquid chromatography (HPLC) on an Agilent 1100 system equipped with an diode array detector (DAD), separations being accomplished on a C18 column, using gradient elution with ammonium acetate buffer and acetonitrile as the mobile phase The HPLC method was optimized and validated, using multiple-specific wavelengths in the visible range to monitor the analytes to provide the highest sensitivity and specificity. The HPLC-DAD developed method proved to be fast (less than 20 minutes run-time), sensitive and accurate and suitable for the routine analysis of soft drinks containing a broad range of colorants. The obtained results showed that: 1) in certain products, besides the declared colorants of the label, several undeclared ones were found; 2) food colorants were generally added in low and rather safe amounts, all samples revealing colorants’ levels in conformity with Romanian and EU regulations.

The popularity of dark chocolate has increased greatly in recent years not only because it is considered a delicatessen, but also due to its high polyphenolic content that provides some interesting healthy properties, such as antioxidant and anti-carcinogenic activities.

This work aims at characterizing and classifying dark chocolate samples based on their geographical origin, cocoa variety and cocoa content using their alkaloid and polyphenolic composition as the data. In the study, 42 chocolate samples from 4 different bean varieties (Criollo, Forastero, Nacional and Trinitario) and 3 different geographical origins (Africa, America and Asia) were analysed in triplicate. Samples were first defatted with chloroform and the resulting residues, containing the polyphenolic species, were further extracted with a methanol/water 60:40 (v:v) solution. The resulting extracts were analysed by high-performance liquid chromatography with UV and fluorescence detection (HPLC-UV-FLD). Compounds were separated on a C18 reversed phase column with an elution gradient using 0.1 % aqueous formic acid solution and acetonitrile as the mobile phase components. Chromatograms were acquired at 280 nm, 325 nm and 370 nm in UV, and at 280 / 330 nm as the excitation/emission wavelengths in FLD.

Principal component analysis (PCA) and partial least squares – discriminant analysis (PLS-DA) were used as exploratory and classification methods, respectively, relying on compositional profiles as the potential descriptors of chocolate features. Classification and discrimination of the chocolates were achieved based on their variety, origin and cocoa content. Results suggested that African samples are characterized by the abundance of flavanols while American samples are richer in alkaloids. Regarding cocoa varieties, (-)-epicatechin, (+)-catechin and procyanidins B2 and C1 are more abundant in Forastero and Nacional derived chocolates, caffeine and theobromine in Criollo, and Trinitario is characterized by the high levels of flavonols and poor (+)-catechin content.

The pandemic caused by the COVID-19 disease has generated different types of social confinement around the world as a strategy to prevent the accelerated spread of the SARS-cov-2 virus. In Mexico, confinement was implemented at the federal level from March 15, 2020 to June 1 of the same year, during that period of time, the key social routines in the establishment of eating habits were modified for most of the population. In the state of Jalisco as in the rest of the country, foods such as eggs, dairy, coffee / tea, sugar, sugary drinks, red meat, sausages, saturated fatty acids (SFA), alcohol, refined cereals and snacks are easily accessible as they are for sale in abundant local distribution centers in most cities and towns of the state.

In this study, residents of the Los Altos area in Jalisco were surveyed regarding changes in the consumption of the aforementioned products during social confinement, as well as data regarding their weight. Our results suggest that in general, there is a significant association between the increase in weight and the modifications to the diet suffered during confinement, also in both women and men, certain foods seem significantly related to weight gain in this period (sugary drinks, refined cereals and SFA). In addition, for the most part, many of the products evaluated were consumed to a greater extent than what is estimated to be consumed per year according to the bibliography; the consequences of this phenomenon have not been studied to date either.

Acorn-fed or bellota dry-cured Iberian ham is a product derived from Iberian pigs fattened up by grazing acorns and pasture in a free-range or montanera regime. It is the most valued animal product from the dehesa agri-ecosystem. Its high commercial value, which reach very different prices depending on the commercial category, has led to an increase in labelling fraud, both in terms of the pig feeding regime and racial purity, causing great damage to the food sector. In this work, a method based on Raman spectroscopy was explored as a rapid in situ screening tool for Iberian ham samples. A total of 110 samples (48 samples of 100% Iberian acorn-fed, 32 samples of 50–75% Iberian acorn-fed and 30 samples of 50–75% Iberian feed-fed dry-cured hams) were analyzed to assess the potential of this technique to differentiate dry-cured Iberian ham four commercial categories: acorn-fed purebred Iberian, acorn-fed crossbred Iberian, free-range feed-fed crossbred Iberian and feed-fed crossbred Iberian. A continuous signal probably due to sample fluorescence was obtained, which hid the Raman scattering signal. Therefore, chemometric treatment was applied in order to extract non-apparent information. High validated classification rates were obtained for feeding regime (83.3%) and breed (86.7%). In addition, an interlaboratory study was carried out to confirm the applicability and robustness of the method, with 52 samples (22 samples of 100% Iberian acorn-fed, 8 samples of 75% Iberian feed-fed and 22 samples of 50% Iberian feed-fed dry-cured hams), obtaining a validated rate above 80%.

Acknowledgments: The authors acknowledge the financial support of the Spanish “Ministerio de Ciencia, Innovación y Universidades” (MICINN) (PGC2018-098363-B-I00), the University of Murcia (R-1328/2018), Fundacion Seneca 19888/GERM/15 and the UCO-FEDER (“Proyectos de I+D+i en el marco del Programa Operativo FEDER Andalucía 2014-2020. Convocatoria 2018”). María García-Nicolás acknowledges a fellowship 21464/FPI/20 from Fundación Séneca, Región de Murcia (Spain).

The amount of plastic waste is continuously increasing being a threat for the environment. Food packaging is the largest source of single-use plastic materials and the majority ends up in landfill/recycling facilities, with some ending up in the environment and the oceans. Plastic recycling is growing significantly to mitigate this problem being key for the sustainability of packaging. However, plastic containers made of recycled materials may contain contaminants from their previous life, collecting system and/or recycling process which may cause a potential threat to consumers’ health, being very important the authentication/characterization of recycled material. A novel non-targeted approach based on static-headspace gas chromatography with mass spectrometry (SHS-GC-MS) has been developed to determine volatile organic fingerprint of plastic packages both virgin and containing different percentages of recycled polyethylene terephthalate (PET). A MS database including 160 volatile organic compounds present in plastic materials according to the literature was created. Among those compounds, seventeen substances were detected and identified by comparing their mass spectra with those of MS database libraries. The combination of the dataset obtained by SHS-GC-MS and the application of chemometrics demonstrated to be a useful tool for the discrimination between virgin and different recycled percentage PET samples as well as for the quantification of recycled plastic content in the PET samples. This analytical tool helps to avoid potential frauds in the recycling plastic field.

Nitrites and nitrates are added to meat products to develop colour, stop the growth of harmful bacteria and improve flavour. However, these additives have some toxicity and could give rise to potentially carcinogenic compounds, such as N-nitrosamines. Traditionally, it was believed that the only responsible for the attractive red colour of meat was myoglobin formed from the addition of nitrite, but new recent trends have shown that the responsible for the red colour of Parma ham is zinc(II) protoporphyrin IX (Zn(II)PPIX). Protoporphyrins are organic compounds consisting of four pyrrole rings linked by methane bridges. The formation of Zn(II)PPIX in ham is unknown and it appears together with other protoporphins biochemically interrelated to it, such as hemin and protoporphyrin XI (PPIX).

The objective of this work is the development of an analytical method for the determination of four protoporphyrins (hemin, heme, PPIX and Zn(II)PPIX) in different type of meat products. The proposed method is carried out by weighing 2 g of ground sample and 1.5 mL of ethyl acetate:acetic acid (3:1) as extractant solvent. The mixture is centrifuged and the extract is filtered and injected (20 µL) in an UHPLC-Q-TOF-MS using electrospray ionization in positive mode. Other detectors such as diode-array and fluorescence were also used. The method was validated by obtaining parameters of linearity, limits of detection and quantification, selectivity, precision and accuracy, and different type of meat products were analysed. Zn(II)PPIX was detected in higher concentration in the meat samples processed in the absence of nitrites, thus demonstrating the inhibition of the formation of this protoporphyrin in the presence of these compounds.

Antioxidants have received much attention in modern society and progressive research have shown that these molecules can help prevent diet-related non-communicable diseases and neurodegenerative diseases by scavenging free radicals, mainly reactive oxygen species, produced during cell metabolism [1][2]. Fruits and vegetables abound in antioxidants and these bioactive molecules are also present in varying quantities in nuts, grains, spices and some meats [3]. Studies have shown that fruits and vegetables increase blood levels of antioxidants significantly and reduce concentrations of markers that are elevated during oxidative stress, such as malondialdehyde and 8-hydroxydeoxyguanosine [4]. Numerous investigations have also reported that daily intake of fruits and vegetables increased the oxygen radical absorption capacity (ORAC) of blood, representing higher antioxidant defence of the body [3]. Though malnutrition has been a challenge in Ghana and Africa as a whole, the potential of many indigenous fruits and vegetables to combat it has not yet been exploited [5]. There is little documentation on their high phytochemical composition and bioactivities hence data on their antioxidant activities is scant [6][7]. In our study, we aimed to investigate the antioxidative potency of 7 wild edible fruits, fruit parts and vegetables of Ghana, namely: Chrysophyllum albidum G. Don, Chrysophyllum perpulchrum Mildbr. ex Hutch. & Dalziel, Chrysophyllum subnudum Baker, Delpydora gracilis A. Chev., Landolphia dulcis var. barteri (Stapf) Pichon, Morinda morindoides (Baker) Milne-Redh.and Sterculia tragacantha Lindl. Samples were collected from the Eastern, Western and Western North regions of Ghana in December 2020. Methanolic extracts were obtained from the dried plant material and the antioxidative activity of each extract was evaluated in vitro using 2,2-diphenyl-1-picrylhydrazyl (DPPH) radical scavenging [8] and oxygen radical absorbance capacity (ORAC) [9] methods. The fruit pulp (mesocarp) of C. subnudum which exhibited the best scavenging activity against the DPPH radical with a remarkable IC50 ± SD value of 45.18 ± 8.61 µg/ml showed an activity of 75.53 ± 8.12 µg/ml in the ORAC assay. Contrastingly, the fruit pulp (mesocarp) of C. albidum demonstrated the best antioxidative activity of 18.54 ± 0.27 µg/ml among the samples in the ORAC assay but recorded a value of 95.63 ± 13.24 µg/ml in the DPPH assay. The seed pulp and epicarp of C. albidum also recorded significant activities of 37.34 ± 6.6 µg/ml and 58.13 ± 3.42 µg/ml respectively in the ORAC assay. Moderate to weak antioxidative effects were observed in the DPPH and ORAC assays for the remaining samples. This study suggests some degree of radical specificity in the scavenging abilities of potent antioxidative fruit parts and provides evidence that the fruit parts of C. albidum and the fruit pulp C. subnudum exhibit significant antioxidant properties when evaluated using both electron and hydrogen atom transfer methods hence they may serve as effective antioxidants which can be used in the food and pharmaceutical industry.

Acknowledgements: Czech University of Life Sciences Prague (project IGA IGA.20213109) supported this research.

CSIR-Plant Genetic Resources Research Institute, Box 7, Bunso, E/R, Ghana

References:

[1] Zafra-Stone S, Yasmin T, Bagchi M, Chatterjee A, Vinson JA, Bagchi D. 2007. Berry anthocyanins as novel antioxidants in human health and disease prevention. Molecular Nutrition & Food Research 51: 675–683.

[2] Bvenura C, Sivakumar D. 2017. The role of wild fruits and vegetables in delivering a balanced and healthy diet. Food Research International 99: 15–30.

[3] Dasgupta A, Klein K. 2014. Fruits, vegetables and nuts: good sources of antioxidants. 209–235. Antioxidants in food, vitamins and supplements. Elsevier, Amsterdam.

[4] Esfahani A, Wong JM, Truan J, Villa CR, Mirrahimi A, Srichaikul K, Kendall CWC. 2011. Health effects of mixed fruits and vegetables concentrates: a systematic review of clinical interventions. Journal of the American College of Nutrition 30: 285-294.

[5] Achaglinkame M, Aderibigbe R, Hensel O, Sturm B, Korese J. 2019. Nutritional characteristics of four underutilized edible wild fruits of dietary interest in Ghana. Foods 8: 1-12.

[6] Stadlmayr B, Charrondière UR, Eisenwagen S, Jamnadass R, Kehlenbeck K. 2013. Nutrient composition of selected indigenous fruits from sub-Saharan Africa. Journal of the Science of Food and Agriculture 93: 2627-2636.

[7] Gyingiri AD, Mills R, Otchere J, Seyram E, Achoribo E, Adu-Kobi Adu-Bobi N, Donkor S, Boatin R, Adom T, Mba Adaboro R, Gomda Y. 2012. Evaluation of the antioxidant potentials of ten leafy vegetable extracts commonly consumed by the Ghanaian population. Electronic Journal of Environmental, Agricultural and Food Chemistry 11: 85-95.

[8] Sharma OP, Bhat TK. 2009. DPPH antioxidant assay revisited. Food Chemistry 113: 1202–1205.

[9] Ou B, Hampsch-Woodhill M, Prior RL. 2001. Development and validation of an improved oxygen radical absorbance capacity assay using fluorescein as the fluorescent probe. Journal of Agricultural and Food Chemistry 49:4619–4626.

The impact of non-Saccharomyces yeasts in fermentation has been investigated during recent years in order to assess the effect of using specific strains individually or in combinations in wine aroma (quality and complexity), as well as in alcohol content. The main purpose of this study was to explore the use of non-Saccharomyces yeasts, individually or in mixed culture with Saccharomyces cerevisiae for the vinification of must originating from the native white wine grape variety Savvatiano. Savvatiano is the most planted grape variety in Greece, cultivated predominantly in Central Greece, and consists the main grape used for the production of flavored wine Retsina. It produces wines with a low aromatic character but well balanced in terms of structure and taste when cultivated in high altitude. Grapes of Savvatiano were harvested during October 2020, were pressed, and the must after cold settlement was inoculated with a. Saccharomyces cereviasiae, b. Metschnikowia pulcherrima and c. mixed culture, in sequential inoculation (M. pulcherrima, followed by S. cereviasiae after 7 days). The progress of fermentations was monitored and the finished wines were analyzed for main wine parameters, as well as sensory attributes by a panel of experts. Wines fermented with S. cerevisiae were characterized by low aromatic intensity on the nose and lack of aromas on the palate, while wines fermented with M. pulcherrima exhibited low scores in general acceptance. However, sequential inoculation with M. pulcherrima, followed by S. cereviasiae produced a full-mouthed wine, characterized by aromatic complexity, citrusy aromas, lasting palate aromas, which exhibited high scores in general acceptance. The results of this study provide useful data in order to further explore the effect of mixed cultures use on fermentation of musts originating from native grape varieties with low aromatic profile.

The hop is rich in tannins meanwhile used as a conventional additive in beer industry, but other application is limited. This study investigated the tyrosinase inhibition activity of extracted hop tannins and the associated structure-function activity. The tannins were extracted and subjected to a gel permeation chromatography (GPC), a nuclear magnetic resonance (NMR) and an acid-cleavage coupled HPLC-ESI-MS/MS analysis to obtain the structural information of the tannins. Then tyrosinase inhibition kinetic assays, inductively coupled plasma optical emission spectrometer and antioxidant (ICP-OES), circular dichroism (CD) as well as molecular docking analysis were applied to investigate the inhibition mechanism. Furthermore, the intracellular inhibition ability of hop tannins was assessed with B16-F10 cells. The results indicated that, hop tannins were composed of (epi)catechin as extensional units and (epi)gallocatechin as terminal units and can be classified as prodelphenidins. The tyrosinase inhibition assays showed the hop tannin was 6.4 times more effective than hydroquinone, meanwhile it inhibited the tyrosinase through competitive-noncompetitive mixed way. The tannins were found to bind on the surface of tyrosinase via forming hydrogen bonding and consequently changed the secondary structure of tyrosinase. The fluorescence and antioxidant assay indicated the tannin had both copper ion chelating and antioxidant ability which may also contribute to the inhibition. The intracellular inhibition analysis showed activity of tyrosinase were reduced by 66.67% and melanin production were found reduced by 34.50% while 10uM hop tannins were applied. These results indicated that, the hops are not only important in beer industry, the hop tannins can be also applied as whitening agents in cosmetic industry.