Chemical composition and sensory profiling of coffees treated with asparaginase to decrease acrylamide formation during roasting.

Acrylamide is an amide formed in the Maillard reaction, with asparagine as the primary amino acid precursor. The intake of large amounts of acrylamide has induced genotoxic and carcinogenic effects in hormone-sensitive tissues of animals. The enzime asparaginase is one of the most effective methods for lowering the formation of acrylamide in foods such as potatoes. However, the reported sensory outcomes for coffee have been unsatisfactory so far. This study aimed to produce coffees with reduced levels of acrylamide by treating them with asparaginase while retaining their original sensory and bioactive profiles. Three raw samples of Coffea arabica, including two specialty coffees, and one of Coffea canephora were treated with 1000, 2000, and 3000 ASNU of the enzyme. Asparagine and bioactive compounds (chlorogenic acids-CGA, caffeine, and trigonelline) were quantified in raw and roasted beans by HPLC and LC-MS, while the determination of acrylamide and volatile organic compounds was performed in roasted beans by CG-MS. Soluble solids, titratable acidity, and pH were also determined. Professional cupping by Q-graders and consumer sensory tests were also conducted. Results were analyzed by ANOVA-Fisher, MFA, PCA and Cluster analyses, with significance levels set at p ≤ 0.05. Steam treatment alone decreased acrylamide content by 18.4%, on average, and 6.1% in medium roasted arabica and canefora coffees. Average reductions of 32.5-56.0% in acrylamide formation were observed in medium roasted arabica beans when 1000-3000 ASNU were applied. In the canefora sample, 59.4-60.7% reductions were observed. However, steam treatment primarily caused 17.1-26.7% reduction of total CGA and lactones in medium roasted arabica samples and 13.9-22.0% in canefora sample, while changes in trigonelline, caffeine, and other evaluated chemical parameters, including the volatile profiles were minimal. Increasing enzyme loads slightly elevated acidity. The only sensory changes observed by Q-graders and or consumers in treated samples were a modest increase in acidity when 3000 ASNU was used in the sample with lower acidity, loss of mild off-notes in control samples, and increased perception of sensory descriptors. The former was selected given the similarity in chemical outcomes among beans treated with 2000 and 3000 ASNU loads.

Evaluation of the safety and quality of Brazil nuts (Bertholletia excelsa) using the tools of dna sequencing technology and aflatoxin profile.

Introduction: Brazil nuts (BNs) result from sustainable extraction and are widely exploited in the Amazon region. Due to the production characteristics in the forest and the nutritional characteristics of these nuts, the occurrence of fungal contamination and the presence of aflatoxins are extensively discussed in the literature as a great aspect of interest and concern. This study aims to evaluate the microbial profile through DNA sequencing and amplification of 16S and ITS genes for bacterial and fungal analysis, respectively, and the presence of mycotoxins using high-performance liquid chromatography with fluorescence detection (HPLC-FD) from different fractions of the nuts processed. Methods: The BN samples, harvest A (HA) and harvest B (HB), from two different harvests were collected in an extractive cooperative in the Amazon region for microbiological analysis (from DNA extraction and amplification of 16S genes, bacteria analysis, and ITS for fungi) and mycotoxins (aflatoxins AFB1, AFB2, AFG1, and AFG2) using HPLC-FD/KobraCell®. Results and discussion: The samples showed a very different microbiome and aflatoxin profile. Genera such as Rothia (HA) and Cronobacter (HB) were abundant during the analysis of bacteria; as for fungi, the genera Aspergillus, Fusarium, Penicillium, and Alternaria were also considered prevalent in these samples. Soil microorganisms, including those pathogenic and related to inadequate hygienic-sanitary production practices, as well as aflatoxins, were found in the samples. However, they were within the established limits permitted by Brazilian legislation. Nuts have a diverse microbiota and are not restricted to fungi of the genus Aspergillus. The microbiological and toxicological profile can vary significantly within the same nut in the same extraction region and can be exacerbated by global climate changes. Therefore, it is necessary to advance sanitary educational actions by applying good production practices and inspection programs to ensure the sustainability and quality of the BN production chain.

SPME-GC-MS untargeted metabolomics approach to identify potential volatile compounds as markers for fraud detection in roasted and ground coffee.

Roasted ground coffee has been intentionally adulterated for economic revenue. This work aims to use an untargeted strategy to process SPME-GC-MS data coupled with chemometrics to identify volatile compounds (VOCs) as possible markers to discriminate Arabica coffee and its main adulterants (corn, barley, soybean, rice, coffee husks, and Robusta coffee). Principal Component Analysis (PCA) showed the difference between roasted ground coffee and adulterants, while the Hierarchical Clustering of Principal Components (HCPC) and heat map showed a trend of adulterants separation. The partial Least-Squares Discriminant Analysis (PLS-DA) approach confirmed the PCA results. Finally, 24 VOCs were putatively identified, and 11 VOCs are candidates for potential markers to detect coffee fraud, found exclusively in one type of adulterant: coffee husks, soybean, and rice. The results for possible markers may be suitable for evaluating the authenticity of ground-roasted coffee, thus acting as a coffee fraud control and prevention tool.

Acute Effects of Coffee Consumption on Blood Pressure and Endothelial Function in Individuals with Hypertension on Antihypertensive Drug Treatment: A Randomized Crossover Trial.

INTRODUCTION: Coffee is a complex brew that contains several bioactive compounds and some of them can influence blood pressure (BP) and endothelial function (EF), such as caffeine and chlorogenic acids (CGAs). AIM: This study aimed to evaluate the acute effects of coffee on BP and EF in individuals with hypertension on drug treatment who were habitual coffee consumers. METHODS: This randomized crossover trial assigned 16 adults with hypertension to receive three test beverages one week apart: caffeinated coffee (CC; 135 mg caffeine, 61 mg CGAs), decaffeinated coffee (DC; 5 mg caffeine, 68 mg CGAs), and water. BP was continuously evaluated from 15 min before to 90 min after test beverages by digital photoplethysmography. Reactive hyperemia index (RHI) assessed by peripheral arterial tonometry evaluated EF before and at 90 min after test beverages. At the same time points, microvascular reactivity was assessed by laser speckle contrast imaging. Repeated-measures-ANOVA evaluated the effect of time, the effect of beverage, and the interaction between time and beverage (treatment effect). RESULTS: Although the intake of CC produced a significant increase in BP and a significant decrease in RHI, these changes were also observed after the intake of DC and were not significantly different from the modifications observed after the consumption of DC and water. Microvascular reactivity did not present significant changes after the 3 beverages. CONCLUSION: CC in comparison with DC and water neither promoted an acute increase in BP nor produced an improvement or deleterious effect on EF in individuals with hypertension on drug treatment who were coffee consumers.

Hybrid Fruits for Improving Health-A Comprehensive Review.

Several species of hybrid fruits, such as citrus, grapes, blueberries, apples, tomatoes, and lingonberries among others, have attracted scientific attention in recent years, especially due to their reported antioxidant and anti-inflammatory properties. The bagasse, leaves, bark, and seeds of these hybrid fruits have large amounts of polyphenols, such as flavonoids, which act as potent antioxidants. Several studies have been carried out in cellular models of neurotoxicity of the extract of these fruits, to document the beneficial effects for human health, as well as to prove its antiproliferative effect in cancer cells. In the present review, through a synthesis of existing information in the scientific literature, we demonstrate that hybrid fruits are a source of antioxidant and bioactive compounds, which act in the inhibition of diseases such as cancer, diabetes, and inflammatory and neurodegenerative diseases, and consequently improving human health.

Anthracnose Controlled by Essential Oils: Are Nanoemulsion-Based Films and Coatings a Viable and Efficient Technology for Tropical Fruit Preservation?

Post-harvest diseases can be a huge problem for the tropical fruit sector. These fruits are generally consumed in natura; thus, their integrity and appearance directly affect commercialization and consumer desire. Anthracnose is caused by fungi of the genus Colletotrichum and affects tropical fruits, resulting in lesions that impair their appearance and consumption. Antifungals generally used to treat anthracnose can be harmful to human health, as well as to the environment. Therefore, essential oils (EO) have been investigated as natural biofungicides, successfully controlling anthracnose symptoms. The hydrophobicity, high volatility, and oxidative instability of essential oils limit their direct application; hence, these oils must be stabilized before food application. Distinct delivery systems have already been proposed to protect/stabilize EOs, and nanotechnology has recently reshaped the food application limits of EOs. This review presents robust data regarding nanotechnology application and EO antifungal properties, providing new perspectives to further improve the results already achieved in the treatment of anthracnose. Additionally, it evaluates the current scenario involving the application of EO directly or incorporated in films and coatings for anthracnose treatment in tropical fruits, which is of great importance, especially for those fruits intended for exportation that may have a prolonged shelf life.

A review of in vitro methods to evaluate the bioaccessibility of phenolic compounds in tropical fruits.

International guidelines strongly advise about the frequent and varied intake of plant in diet. In this scenario, the consumption of fruits is closely related to health benefits due to the abundant presence of bioactive substances. Accordingly, the production of tropical fruits has stood out worldwide, reaching records since the past decade. However, to ensure that phenolic substances are indeed used by the body, they need to be accessible for absorption. For this purpose, several methods are used to assess the phenomenon of bioaccessibility. We provide information on i) in vitro methods for the evaluation of the bioaccessibility of phenolic compounds in tropical fruits, including their derivatives and by-products; ii) a study performed using a semi-dynamic in vitro digestion model; iii) simulated digestion with a dialysis membrane step, polyphenol transport/uptake using cell culture, and in vitro colonic fermentation process. Although standardized static and semi-dynamic in vitro digestion methods already exist, few studies use these protocols to assess the bioaccessibility of polyphenols in tropical fruits. To guarantee that in vitro digestion assays reproduce consistent results compared to in vivo reference methods, it is essential to universalize standardized methods that allow the comparison between results, enabling the validation of in vitro digestion methods.

Molecular Mechanisms of Coffee on Prostate Cancer Prevention.

Prostate cancer (PCa) is one of the most common types of cancer among men, and coffee is associated with a reduced risk of developing PCa. Therefore, we aim to review possible coffee molecular mechanisms that contribute to PCa prevention. Coffee has an important antioxidant capacity that reduces oxidative stress, leading to a reduced mutation in cells. Beyond direct antioxidant activity, coffee stimulates phase II enzymatic activity, which is related to the detoxification of reactive metabolites. The anti-inflammatory effects of coffee reduce tissue damage related to PCa development. Coffee induces autophagy, regulates the NF-κB pathway, and reduces the expression of iNOS and inflammatory mediators, such as TNF-α, IL-6, IL-8, and CRP. Also, coffee modulates transcriptional factors and pathways. It has been shown that coffee increases testosterone and reduces sex hormone-binding globulin, estrogen, and prostate-specific antigen. Coffee also enhances insulin resistance and glucose metabolism. All these effects may contribute to protection against PCa development.

Evaluation of the antioxidant capacity, volatile composition and phenolic content of hybrid Vitis vinifera L. varieties sweet sapphire and sweet surprise.

Bioactive compounds were extracted using two different extraction solvents (acetone and water) from pulp and whole grape berries derived from hybrid Vitis vinifera L. varieties Sweet sapphire (SP) and Sweet surprise (SU) and were characterised based on a comprehensive metabolomic approach by chromatography coupled with mass spectrometry (UPLC-QTOF-MSE and GC-FID/MS). GC-FID/MS analysis was performed with two different extraction methods (solvent extraction method and solid-phase extraction). Anthocyanins were characterised and quantified by HPLC-UV. The antioxidant potential was assessed by different assays. SP acetone extract from grape skin had the highest mean to DPPH, FRAP, ORAC and phenolic content SP samples, also showed higher anthocyanin content. Globally, 87 phenolic compounds were identified. The relative quantification by UPLC-MSE showed flavonoids the most abundant class. Forty two compounds were found in the volatile fraction of SU, while only thirty one volatile compounds were found in the SP samples.

Ochratoxin a levels in fermented specialty coffees from Caparaó, Brazil: Is it a cause of concern for coffee drinkers?

Although postharvest coffee fruit fermentation can improve coffee flavour and quality, the mycotoxin ochratoxin A (OTA) can also be a result of microbiological activity, albeit in the later drying step of coffee processing. To evaluate the possible occurrence of OTA contamination in postharvest fruit fermentation, fourteen coffees that entailed two different postharvest fruit fermentation times were evaluated. These coffees originated in the surroundings of the village of Pedra Menina in the qualified Denomination of Origin and coffee producer region of Caparaó on the border between Minas Gerais and Espírito Santo states in Brazil. All coffees were classified according to the Specialty Coffee Association (SCA) protocol and 12 achieved specialty level. OTA was determined in all 14 coffees using immunoaffinity for sample clean-up and high-performance liquid chromatography with fluorescence detection for quantification. One sample presented an OTA concentration of 0.75 µg kg-1 and two samples showed OTA concentrations of 0.87 µg kg-1. The other samples had concentrations of OTA below the limit of quantification obtained in this work (0.64 µg kg-1). Thus, all samples showed OTA concentrations far below the most stringent maximum residue limit (MRL) of 5 µg kg-1 established for roasted coffees by European legislation. These low levels were similar to most of the previous results for Brazilian coffees listed and tabled in this work. This comparison showed that OTA contamination due to this kind of postharvest process - fruit fermentation - should not be a concern for producers and consumers of these fermented coffees.

Antitumor Effects of Freeze-Dried Robusta Coffee (Coffea canephora) Extracts on Breast Cancer Cell Lines.

Coffee consumption is believed to have chemopreventive and chemotherapeutic effects and to contribute to preventing the development and progression of cancer. However, there is still controversy around these claims. As indicated in our previous works, diet can influence the risk of breast cancer. Intake of coffee is hypothesized to reduce this risk, but current scientific evidence is not conclusive. This work is aimed at studying the effects of Robusta coffee bean extract on cell viability, proliferation, and apoptosis of different human cancers, especially breast cancer cell lines. To this end, cell viability was evaluated by Alamar Blue in 2D and 3D models, the cell cycle by PI, apoptosis by annexin V, mitochondrial morphology, and functionality by mitoTracker, and colony formation capacity by the clonogenic assay. Green and dark coffee extract significantly reduced viability in human breast, colorectal, brain, and bone cancer cells. Coffee anticancer activity was clearly evidenced in MDA-MB-231 (ER-) and MCF-7 (ER+) breast cancer cells but not in the normal breast cell line. In addition, coffee extract induces an increase S phase and a decrease G2/M population in breast cancer cells, affected the mitochondrial morphology, and triggered apoptosis. MDA-MB-231 breast cancer cells lost their clonogenic capacity after treatment. The antitumor activity was demonstrated in both 2D and 3D culture cell models.

Brazilian Coffee Production and the Future Microbiome and Mycotoxin Profile Considering the Climate Change Scenario.

Brazil holds a series of favorable climatic conditions for agricultural production including the hours and intensity of sunlight, the availability of agricultural land and water resources, as well as diverse climates, soils and biomes. Amidst such diversity, Brazilian coffee producers have obtained various standards of qualities and aromas, between the arabica and robusta species, which each present a wide variety of lineages. However, temperatures in coffee producing municipalities in Brazil have increased by about 0.25 °C per decade and annual precipitation has decreased. Therefore, the agricultural sector may face serious challenges in the upcoming decades due to crop sensitivity to water shortages and thermal stress. Furthermore, higher temperatures may reduce the quality of the culture and increase pressure from pests and diseases, reducing worldwide agricultural production. The impacts of climate change directly affect the coffee microbiota. Within the climate change scenario, aflatoxins, which are more toxic than OTA, may become dominant, promoting greater food insecurity surrounding coffee production. Thus, closer attention on the part of authorities is fundamental to stimulate replacement of areas that are apt for coffee production, in line with changes in climate zoning, in order to avoid scarcity of coffee in the world market.

Bioactive compounds, antioxidant activity and antiproliferative effects in prostate cancer cells of green and roasted coffee extracts obtained by microwave-assisted extraction (MAE).

Coffee consumption has been investigated as a protective factor against prostate cancer. Coffee may be related to prostate cancer risk reduction due to its phytochemical compounds, such as caffeine, chlorogenic acids, and trigonelline. The roasting process affects the content of the phytochemicals and undesired compounds can be formed. Microwave-assisted extraction is an alternative to conventional extraction techniques since it preserves more bioactive compounds. Therefore, this study aimed to evaluate the phytochemical composition and the putative preventive effects in prostate cancer development of coffee beans submitted to four different coffee-roasting degrees extracted using microwave-assisted extraction. Coffea arabica green beans (1) were roasted into light (2), medium (3) and dark (4) and these four coffee samples were submitted to microwave-assisted extraction. The antioxidant capacity of these samples was evaluated by five different methods. Caffeine, chlorogenic acid and caffeic acid were measured through HPLC. Samples were tested against PC-3 and DU-145 metastatic prostate cancer cell lines regarding their effects on cell viability, cell cycle progression and apoptotic cell death. We found that green and light roasted coffee extracts had the highest antioxidant activity. Caffeine content was not affected by roasting, chlorogenic acid was degraded due to the temperature, and caffeic acid increased in light roasted and decreased in medium and dark roasted. Green and light roasted coffee extracts promoted higher inhibition of cell viability, caused greater cell cycle arrest in S and G2/M and induced apoptosis more compared to medium and dark roasted coffee extracts and the control samples. Coffee extracts were more effective against DU-145 than in PC-3 cells. Our data provide initial evidence that among the four tested samples, the consumption of green and light coffee extracts contributes to inhibit prostate cancer tumor progression features, potentially preventing aspects related to advanced prostate cancer subtypes.

Near-Infrared Spectroscopy Applied to the Detection of Multiple Adulterants in Roasted and Ground Arabica Coffee.

Roasted coffee has been the target of increasingly complex adulterations. Sensitive, non-destructive, rapid and multicomponent techniques for their detection are sought after. This work proposes the detection of several common adulterants (corn, barley, soybean, rice, coffee husks and robusta coffee) in roasted ground arabica coffee (from different geographic regions), combining near-infrared (NIR) spectroscopy and chemometrics (Principal Component Analysis-PCA). Adulterated samples were composed of one to six adulterants, ranging from 0.25 to 80% (w/w). The results showed that NIR spectroscopy was able to discriminate pure arabica coffee samples from adulterated ones (for all the concentrations tested), including robusta coffees or coffee husks, and independently of being single or multiple adulterations. The identification of the adulterant in the sample was only feasible for single or double adulterations and in concentrations ≥10%. NIR spectroscopy also showed potential for the geographical discrimination of arabica coffees (South and Central America).

Use of asparaginase for acrylamide mitigation in coffee and its influence on the content of caffeine, chlorogenic acid, and caffeic acid.

A factorial design with a duplicate in the central point was used to investigate the effect of treating arabica coffee beans with asparaginase. The investigated factors were enzymatic load (1000 and 5000 ASNU/Kg), water percentage (30 and 90%), and hydrolysis time (1 and 3 h). The acrylamide content was determined by UPLC-MS/MS, and the caffeic acid, chlorogenic acid and caffeine concentrations were determined by HPLC-DAD. The statistical analysis was carried out in the R platform using RStudio graphical interface. The results indicated the importance of coffee bean pretreatment with steam, and that the enzyme load reduced the acrylamide content to 65 mg/kg in coffee beans. The predicted reduction was obtained with hydrolysis time of 2 h, water content of 90%, and asparaginase load of 5000 ASNU/kg. The asparaginase treatment did not influence the major bioactive compounds in coffee.

Potential use of vegetable proteins to reduce Brazil nut oil oxidation in microparticle systems.

Brazil nut oil is mostly composed of unsaturated fatty acids, some of which are associated with decreased incidence of cardiovascular diseases. Vegetable proteins have been increasingly used as wall material for partial replacement of carbohydrates and whey proteins. In order to create an oil preservation method, Brazil nut oil was encapsulated with three different types of vegetable protein concentrates and gum arabic (GA): rice (RPC + GA); pea (PPC + GA); and soy (SPC + GA) .For this purpose, vegetable protein concentrates were characterized, and after the drying process the physicochemical characteristics of the microparticles were evaluated. The most stable emulsion, after seven days of evaluation, was composed of RPC + GA. RPC + GA. This treatment was also more stable based on the shelf life assessments. We concluded that RCP microparticles were the best option for encapsulating Brazil nut oil in comparison with the other particles evaluated. In addition, the product obtained is potentially capable of being included in various processed foods.

Fruit-based drink sensory, physicochemical, and antioxidant properties in the Amazon region: Murici (Byrsonima crassifolia (L.) Kunth and verbascifolia (L.) DC) and tapereba (Spondia mombin).

Increased fruit consumption due its protective effect on the organism is accompanied by the development of the processing industry of these products. The aim of this work was to optimize fruit pulp-based beverage formulations from the murici and tapereba Amazon region, taking into account their sensory acceptance and antioxidant activity. Total soluble solid content, reducing sugar content, titratable acidity contents, pH, and ascorbic acid content were determined in pulps and formulations. The total content phenolic compounds and antioxidant activity were also evaluated. A 22 factorial experiment was formulated to optimize ingredients for the production of murici and tapereba fruit drinks. The murici pulp had higher acidity and higher ascorbic acid content. The analysis of phenolic compounds and antioxidant activity presented higher quantity in tapereba pulp. Tapereba-based beverages had better acceptance by the evaluated criteria. Fruit-based beverages murici and tapereba are a well-accepted product and have important nutritional characteristics.

Cellulose nanocrystals from grape pomace and their use for the development of starch-based nanocomposite films.

Nanocomposite films prepared from starch (ST) in the presence of cellulose nanocrystals (CNCs) was performed using grape pomace as raw material. CNCs were obtained by acid hydrolysis and added to filmogenic solutions (1, 2, 5, 10 and 15 g/100 g of ST). Cellulose, CNCs and Nanocomposites were characterized. Amorphous non-cellulosic materials were removed from the grape pomace presented values for CrI 64% and 71% and yield 12 and 70% in Cellulose and CNCs, respectively. Nanocomposites showed smaller permeability and the addition of 5 to 15% CNCs formed more opaque films and had improved tensile strength and Young's modulus. The addition of CNCs from 5 to 15% proved to be effective in improving mechanical properties and decreasing water vapor permeability, important characteristics in food packaging materials. This study provided an effective method to obtain CNCs from the agroindustrial waste and open the way to produce high-value starch based nanocomposites.

Effect of the roasting levels of Coffea arabica L. extracts on their potential antioxidant capacity and antiproliferative activity in human prostate cancer cells.

Coffee, besides being one of the most consumed stimulating beverages in the world, has important bioactive activities, which have been attracting increasing attention from researchers. However, the standard process of roasting causes changes in its chemical composition. In the present study, extracts obtained from green and roasted beans (light, medium and dark) of Coffea arabica Linnaeus were submitted to high-power ultrasonic extraction and atomization by spray drying. Colorimetric analysis was used to classify the roasting levels of the dried extract samples. The effects of the roasting process on the bioactivity of the dried extracts were verified through the following assays: caffeine, chlorogenic acid and caffeic acid, by HPLC-PDA; total phenolics by Folin-Ciocalteu; antioxidant activity by DPPH, FRAP, ABTS and ORAC; antiproliferative activity, using the MTT assay; and cell cycle and apoptosis by flow cytometry in metastatic prostate cancer cell lines from bone (PC-3) and brain (DU-145). The results showed that the lowest levels of caffeine, chlorogenic and caffeic acids were observed in dark roasted coffee. In comparison to medium and dark extracts in PC-3 cells, the green and light coffee extracts had higher antioxidant activities and promoted cytotoxicity followed by cell cycle arrest in phase S and apoptosis induction. Thus, the roasting level of the coffee extracts may be related to the potential chemoprotective effects of Coffea arabica L. in prostate cancer cells.

Production and characterization of Gluconacetobacter xylinus bacterial cellulose using cashew apple juice and soybean molasses.

Bacterial cellulose (BC) has been largely used in biomedical and technological fields. The use of agro-industrial byproducts as alternative source of carbon and nitrogen in culture media reduces the BC cost production, adds value to the byproducts and minimizes the environmental impact. In this study, the use of cashew apple juice and soybean molasses were evaluated to produce BC by Gluconacetobacter xylinus in comparison to the usual Hestrin and Schramm medium (HS). BC produced in static cultivation was characterized by X-ray diffraction, Fourier transform infrared spectroscopy and thermogravimetric analysis. The BC production (4.50 g L-1) obtained from the medium using cashew apple juice as carbon source (20 g L-1) with soybean molasses as nitrogen source (10 g L-1) was superior than HS medium (4.03 g L-1). Morphological analysis showed that bacterial celluloses produced with agro-industrial byproducts combined were similar to those found for the pellicle obtained from HS medium.