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BACKGROUND:Lower risk of digestive tract cancer development has been associated with polyphenol intake. Bactris guineensis is an edible endemic palm that grows in Central and South America. OBJECTIVE:This study performs a phenolic characterization of Bactris guineensis and evaluates the bioactivity of this fruit. METHODS:The phenolic compounds of B. guineensis were characterized by HPLC-UV-HRMS analyses and the antioxidant activity was measured by chemical and cellular methods. Additionally, cytotoxicity of B. guineensis polyphenols was performed on 4 cancer cell lines and the pro-apoptotic effect was evaluated by flow cytometry using annexin staining. RESULTS:The major phenolic compounds of B. guineensis were proanthocyanidins. The extract IC50 for DPPH was 3.3±0.2 μg/mL and for induced intracellular ROS was 153±13 μg/mL. MTT cytotoxic assays demonstrate IC50 values between 16.6 and 24.9 μg/mL for the colon and hepatic adenocarcinomas, with high selectivity effects towards cancer cells compared to non-tumor cells. A 20 to 50% early apoptotic effect was observed in cancer cells lines by Annexin/PI staining. CONCLUSIONS:B. guineensis evidenced an important radical scavenging activity and a strong cytotoxic activity against hepatic and colorectal carcinoma cells, showing better values than procyanidin extracts from other fruits previously described. Universidad de Costa Rica/[422-B7-099]/UCR/Costa Rica UCR::Vicerrectoría de Investigación::Unidades de Investigación::Ciencias Agroalimentarias::Centro Nacional de Ciencia y Tecnología de Alimentos (CITA) UCR::Vicerrectoría de Investigación::Unidades de Investigación::Ciencias Agroalimentarias::Centro de Investigación en Nutrición Animal (CINA) UCR::Vicerrectoría de Docencia::Salud::Facultad de Farmacia UCR::Vicerrectoría de Docencia::Salud::Facultad de Medicina::Escuela de Medicina

Honey is a food produced from honey bee widely used for the sweetening power and for its biological properties. In order to prevent the infection of the hive, different xenobiotics (antibiotics, pesticide) were frequently employed. One of these substances is the chloramphenicol, that given its chemical stability could often found in food. Chloramphenicol have several side effects in humans after their ingestion and for this reason their intake must be avoid. The aim of this study, was developed an ultrasound-assisted dispersive liquid-liquid microextraction method coupled with UHPLC MS/MS determination, for fast and accurate analysis of chloramphenicol in honey. The parameters affecting on extraction efficiency were carefully optimized using an experimental design in order to maximized the recovery reducing matrix effects. After the optimization the method was validated and successfully applied to 66 honey samples.

Psoriasis is an inflammatory disease of the epidermis based on an immunological mechanism involving Langerhans cells and T lymphocytes that produce pro-inflammatory cytokines. Genetic factors, environmental factors, and improper nutrition are considered triggers of the disease. Numerous studies have reported that in a high number of patients, psoriasis is associated with obesity. Excess adipose tissue, typical of obesity, causes a systemic inflammatory status coming from the inflammatory active adipose tissue; therefore, weight reduction is a strategy to fight this pro-inflammatory state. This study aimed to evaluate how a nutritional regimen based on a ketogenic diet influenced the clinical parameters, metabolic profile, and inflammatory state of psoriasis patients. To this end, 30 psoriasis patients were subjected to a ketogenic nutritional regimen and monitored for 4 weeks by evaluating the clinical data, biochemical and clinical parameters, NMR metabolomic profile, and IL-2, IL-1β, TNF-α, IFN-γ, and IL-4 concentrations before and after the nutritional regimen. Our data show that a low-calorie ketogenic diet can be considered a successful strategy and therapeutic option to gain an improvement in psoriasis-related dysmetabolism, with significant correction of the full metabolic and inflammatory status.

This study reports a fast and automated analytical procedure based on an on-line SPE-HPLC–MS/MS method for the automatic pre-concentration, clean up and sensitive determination of OTA in wine. The amount of OTA contained in 100 μL of sample (pH ≅ 5.5) was retained and concentrated on an Oasis MAX SPE cartridge. After a washing step to remove matrix interferents, the analyte was eluted in back-flush mode and the eluent from the SPE column was diluted through a mixing Tee, using an aqueous solution before the chromatographic separation achieved on a monolithic column. The developed method has been validated according to EU regulation N. 519/2014 and applied for the analysis of 41 red and 17 white wines. The developed method features minimal sample handling, low solvent consumption, high sample throughput, low analysis cost and provides an accurate and highly selective results

Food processing industry is accompanied with the generation of a great production of wastes and by-products exceptionally rich in bioactive compounds (especially phenolics), with antioxidant activity. The recovery of these health molecules constitutes a key point for the valorization of by-products, with the possibility of creating new ingredients to be used for the formulation of food and cosmetic products. One of the main limitations to reuse by-products is linked to the high cost to obtain bioactive compounds, consequently in order to exploit these resources commercially valuable it is necessary to develop innovative, economic and environmentally friendly extraction strategies. These extraction methods should be able to reduce petroleum solvents, energy consumption and chemical wastes, protecting both environment and consumers and ensuring safe and high-quality final products. The purpose of this review is to summarize current knowledge and applications of the new extraction techniques such as supercritical fluid extraction, pressurized liquid extraction, ultrasound assisted extraction applied to polyphenols extraction from agricultural food by-products. Particular attention has been paid to theoretical background, highlighting mechanisms and safety precautions. Authors concluded that relevant results of these techniques represent an opportunity to industrial scale-up, improving the extraction yields, minimizing time, costs and environmental impact.

Distillation wastewaters (DWWs) are generated during the essential oil steam distillation from aromatic herbs. Despite of growing interest on novel source of natural antioxidant compounds as food additives, studies on DWWs are scarse. Herein, the potential of DWWs produced by the distillation of packaged fresh basil, rosemary and sage wastes was evaluated by chemical and antioxidant characterization. HPLC-DAD-HRMS profiling revealed that DWWs contain water-soluble phenolic compounds, mainly caffeic acid derivatives and flavonoid glycosides, with rosmarinic acid (RA) as predominant components (29–135 mg/100 mL). DWWs demonstrated high levels of total phenolic compounds (TPC, 152–443 mg GAE/100 mL) and strong antioxidant capacities, in ORAC, DPPH and ABTS assays (1101–4720, 635–4244 and 571–3145 μmol TE/100 mL, respectively). Highly significant correlations of TEAC values with TPC and RA contents revealed that phenolic compounds and high RA content were responsible of DWWs antioxidant properties.Thus, DWWs are proposed as a new promising source of natural food additives and/or functional ingredients for cosmetic, nutraceutical and food applications

Consumers are aware of diet causing health problems and therefore there is an increased demand for natural ingredients that are expected to be safe and health-promoting. Many of these compounds belong to the class of flavonoids and can be divided into these five groups: flavanones, flavones, flavonols, flavanols, isoflavones and anthocyanidins. Extracts from citrus fruits are usually used as functional ingredients for several products. The aim of this paper was to develop an UHPLC-UV-ESI-HRMS method to define the metabolite profile of different parts of citrus fruit, of a particular cultivar called 'Ovale Calabrese', and in its main by-products. The high resolution mass spectrometry analysis allowed the identification of 27 compounds belonging to the classes of flavonoids and terpenoids. The high contents of phytochemical compounds, reveal the potential use of the 'Ovale Calabrese' as a rich source of nutraceutical compounds.

Propolis is an attractive natural ingredient to design health products due to its pharmacological effects. Our chemical investigation of a polar extract of Nigerian propolis (NP) led the isolation and identification of five isoflavonoids (1-4, 6), one diarylpropane (5) and one prenylated flavanone (7) by the combination of chromatographic and spectroscopic techniques. Compounds 1, 4 and 7 were found to be the main markers in NP (8.0, 5.0 and 4.0 mg/g of dry extract, respectively). Moreover, NP and its phenolic constituents exhibited in vitro free radical scavenging activity together with a promising antidiabetic effect against α-amylase and α-glucosidase enzymes. Finally, NP showed also a moderate inhibition of Helicobacter pylori growth. These results suggested that NP could be a good candidate in nutraceuticals and food products.

Roasted hazelnut skins (RHS) represent a byproduct of kernel industrial processing. In this research, a RHS extract (RHS-M) and its fraction RHS-M-F3 enriched in proanthocyanidins (PAs), with antioxidant activity, were characterized in terms of total phenolic compound and PA contents. RHS-M and RHS-M-F3 showed antifungal properties against Candida albicans SC5314 (MIC2 = 3.00 and 0.10 μg/mL and MIC0 = 5.00 and 0.50 μg/mL, respectively), determined by the microbroth dilution method and Candida albicans morphological analysis. No cytotoxic effect on HEKa and HDFa cell lines was exhibited by RHS-M and RHS-M-F3. The metabolite profiling of RHS-M and RHS-M-F3 was performed by thiolysis followed by HPLC-UV-HRMS analysis and a combination of HRMS-FIA and HPLC-HRMS(n). Extract and fraction contain oligomeric PAs (mDP of 7.3 and 6.0, respectively, and DP up to 10) mainly constituted by B-type oligomers of (epi)-catechin. Also, (epi)-gallocatechin and gallate derivatives were identified as monomer units, and A-type PAs were detected as minor compounds.

Beer is one the most consumed alcoholic beverage in the world and its contamination with mycotoxins is of public health concern. This study reports a fast and automated analytical procedure based on a multi-heart-cutting two-dimensional liquid chromatography tandem mass spectrometry method using electrospray ionization for the determination of seven mycotoxins (aflatoxins B1, B2, G2 and G1, ochratoxin A, fumonisins B1 and B2) in beers. The developed method was based on the heart-cutting 2D- HPLC technique in which only the specific portions of the first dimension, in the retention time of analytes, were transferred into the second dimension for the further separation and successive determination. The method uses two different chromatographic columns; in the first dimension, 50 μL of sample was injected on first column, and mycotoxins elution regions were collected in a loop and transferred into the second column for the separation of analytes. Each column operated in gradient elution mode in order to eliminate interfering compounds and improve separation and peak shape. After the optimization, the method has been validated according to EU regulation and finally applied for the analysis of forty beer samples collected from Italian supermarkets. Among all mycotoxins studied, fumonisins B1 was the most widely distributed in analysed beers (>21%) in the range from 0.6 to 12.3 ng mL−1. The automated methodology developed was able to determine accurately and simultaneously seven mycotoxins in beer. This provided a significant reduction of sample handle and, consequently of analysis time.

Food industry produces a large amount of onion wastes. Due to the high amount of bioactive compounds in onion by-products an idea for their reuse, could be use them as source of high-value functional and health ingredients. In this study, outer dry layers of coppery onion “Ramata di Montoro” were used as source of bioactive compounds. Firstly, the chemical profile of secondary metabolites of exhaustive extract, obtained by ultrasound assisted extraction was established by UHPLC-UV-HRMS/MS analysis. Subsequently, the supercritical fluid extraction was used as alternative and green method to recover flavonoids from onion skin. Main parameters such as pressure, temperature and composition of solvent modifier were optimized in order to improve the extraction efficiency of SFE technique, by using a response surface Box–Behnken design.

Artichoke by-products are a suitable source of health-promoting ingredients for the production of dietary supplements and food additives. A pressurized hot water extraction (PHWE) was developed to recover caffeoylquinic acids (CQAs) and flavone glycosides (FLs) from agro-industrial artichoke by-products. The main factors influencing PHWE efficiency and CQA isomerization (temperature, numbers of cycles, modifier, and extraction time) were carefully studied and optimized by response surface design. The proposed PHWE procedure provides an exhaustive extraction of CQAs and FLs (recoveries: 93–105% and 90–105%) from artichoke external bracts and leaves of different cultivars (p>0.05), without significant formation of artefacts generated by high temperatures. PHWE extracts showed CQA and FL levels (14–37 mg/g and 3–19mg/g, respectively) comparable to commercial products and marked antioxidative effects (EC50 11–83μg/mL) by cellular antioxidant activity assay in human hepatocarcinoma HepG2 cells. These results proved that PHWE is an excellent green technique to recover bioactive compounds from artichoke agro-industrial residues

The occurrence of aflatoxin M1 in 69 milk samples collected in a south region of Italy in 2016 was evaluated. The samples were analysed using an automated method based on online SPE coupled with UHPLC tandem mass spectrometry. After a salt induced liquid–liquid extraction with acetonitrile to remove protein from milk, the extract was diluted with water and analysed using an automated online SPE MS/MS method. Among the analysed samples no one had AFM1 higher than the legally allowable limits whereas 71.4% of the other analysed samples were above the LOD of the method. The highest contamination level of AFM1 was found in pasteurised milk (44.39 ng kg−1). The results show the worrying and widespread of AFM1 contamination, highlighting the necessity of monitoring studies in order to evaluate the reduction of the maximum legal limit.

During the essential oil steam distillation from aromatic herbs, huge amounts of distillation wastewaters (DWWs) are generated. These by‐products represent an exceptionally rich source of phenolic compounds such as rosmarinic acid (RA) and caffeic acid (CA). Herein, the alternative use of dried basil DWWs (dDWWs) to perform a selective extraction of RA and CA by pressurized liquid extraction (PLE) employing bio‐based solvent was studied. To select the most suitable solvent for PLE, the theoretical modelling of Hansen solubility parameters (HSP) was carried out. This approach allows reducing the list of candidate to two solvents: ethanol and ethyl lactate. Due to the composition of the sample, mixtures of water with those solvents were also tested. An enriched PLE extract in RA (23.90 ± 2.06 mg/g extract) with an extraction efficiency of 75.89 ± 16.03% employing a water‐ethanol mixture 25:75 (% v/v) at 50°C was obtained. In the case of CA, a PLE extract with 2.42 ± 0.04 mg/g extract, having an extraction efficiency of 13.86 ± 4.96% using ethanol absolute at 50°C was achieved. DWWs are proposed as new promising sources of natural additives and/or functional ingredients for cosmetic, nutraceutical, and food applications., The present work was supported by the projects AGL2014-53609-P (Ministerio de Economia y Competitividad, Spain), S2013/ABI-2728 (Comunidad de Madrid) and I-LINK1096 (CSIC).

A molecularly imprinted polymer was prepared using (E)-resveratrol as template and was evaluated for multicomponent multiclass analysis of polyphenolic compounds in complex matrices such as natural and alcoholic beverages. Chromatographic evaluation of the polymer exhibited high selectivity for (E)-resveratrol and its structural analogues, quercetin, and other flavonoids. An analytical procedure based on molecularly imprinted solid phase extraction (MISPE) and high-performance liquid chromatography coupled to UV detector was developed and validated for determination of (E)-resveratrol and quercetin in wine and fruit juice samples. The specific binding capacity of the MIP was estimated as 80 μg g(-1) polymer by the cartridge test. MISPE sample pretreatment allows an excellent sample cleanup, enormously decreasing the number of coextracted potentially interfering compounds. Under the described conditions, by extracting 2 mL samples a clean extract is obtained and (E)-resveratrol and quercetin could be easily identified at concentration levels of, respectively, 1.5 and 7.0 μg L(-1).

This study reports a fast and automated analytical procedure for the analysis of aflatoxin M1 (AFM1) in milk and dairy products. The method is based on the simultaneous protein precipitation and AFM1 extraction, by salt-induced liquid-liquid extraction (SI-LLE), followed by an online solid-phase extraction (online SPE) coupled to ultra-high-pressure-liquid chromatography-tandem mass spectrometry (UHPLC-MS/MS) analysis to the automatic pre-concentration, clean up and sensitive and selective determination of AFM1. The main parameters affecting the extraction efficiency and accuracy of the analytical method were studied in detail. In the optimal conditions, acetonitrile and NaCl were used as extraction/denaturant solvent and salting-out agent in SI-LLE, respectively. After centrifugation, the organic phase (acetonitrile) was diluted with water (1:9 v/v) and purified (1. mL) by online C18 cartridge coupled with an UHPLC column. Finally, selected reaction monitoring (SRM) acquisition mode was applied to the detection of AFM1.Validation studies were carried out on different dairy products (whole and skimmed cow milk, yogurt, goat milk, and powder infant formula), providing method quantification limits about 25 times lower than AFM1 maximum levels permitted by EU regulation 1881/2006 in milk and dairy products for direct human consumption. Recoveries (86-102%) and repeatability (RSD < 3, n=. 6) meet the performance criteria required by EU regulation N. 401/2006 for the determination of the levels of mycotoxins in foodstuffs. Moreover, no matrix effects were observed in the different milk and dairy products studied.The proposed method improves the performance of AFM1 analysis in milk samples as AFM1 determination is performed with a degree of accuracy higher than the conventional methods. Other advantages are the reduction of sample preparation procedure, time and cost of the analysis, enabling high sample throughput that meet the current concerns of food safety and the public health protection.

Artichoke by-products, produced from agricultural procedures and the processing industry, represent a huge amount of discarded material. In this research, the main artichoke by-products, bracts and leaves, were characterized in terms of their bioactive constituents (phenolic compounds and inulin) and cellular antioxidant potential to estimate their nutraceutical potential. The ultrahigh-performance-ultraviolet detection-high resolution mass spectroscopy (UHPLC-UV-HRMS) profiles of both by-products show that 5-caffeoylquinic acid and 1,5-dicaffeoylquinic acid are the most abundant bioactive compounds, and the content of flavone glycosides can be used to discriminate between bracts and leaves. Artichoke by-products contain a remarkable overall phenolic content (0.5-1.7 g per 100 g dry matter), whereas they differ widely in the amounts of inulin with higher levels in bracts (3.8-8.2 g per 100 g dry matter). The cellular antioxidant activities of bract and leaf extracts (half maximal effective concentration (EC50) = 26.6-124.1 mg L-1) are better than or similar to that of a commercial leaf extract, and are related to the dicaffeoylquinic acid levels, particularly to 1,5-dicaffeoylquinic acid. These results reveal that artichoke by-products are a promising and cheap source of bioactive compounds. Bracts could be used as a source of inulin and caffeoylquinic acids for the production of food additives and nutraceuticals and also as an alternative to the traditional application of leaf extracts.

Honey is a valuable functional food rich in phenolic compounds with a broad spectrum of biological activities. Analysis of the phenolic compounds in honey is a very promising tool for the quality control, the authentication and characterization of botanical origin, and the nutraceutical research. This work describes a novel approach for the rapid analysis of five phenolic acids and 10 flavonoids in honey. Phenolic compounds were rapidly extracted and concentrated from diluted honey by dispersive liquid-liquid microextraction (DLLME) and then analyzed using high performance liquid chromatography with UV absorbance detection (HPLC-UV). Some important parameters, such as the nature and volume of extraction and dispersive solvents, pH and salt effect were carefully investigated and optimized to achieve the best extraction efficiency. Under the optimal conditions, an exhaustive extraction for twelve of the investigated analytes (recoveries >70%), with a precision (RSD