Loading...
58 results
Search Results
Now showing 1 - 10 of 58
- The salivary volatome in breast cancerPublication . Pereira, Jorge A. M.; Taware, Ravindra; Porto-Figueira, Priscilla; Rapole, Srikanth; Câmara, José S.
- Beer volatile fingerprinting at different brewing stepsPublication . Alves, Vera; Gonçalves, João; Figueira, José A.; Ornelas, Laura P.; Branco, Ricardo N.; Câmara, José S.; Pereira, Jorge A. M.Volatile fingerprints of a lager beer were carried out throughout five brewing steps to characterize the changes encompassing this process. Overall, 60 volatile organic metabolites (VOMs) were identified by headspace solid phase microextraction followed by gas chromatography mass spectrometry (HS-SPME/GC–MS). Specific profiles were observed at different brewing steps - aldehydes and furans dominate in wort, whereas the aliphatic esters and alcohols predominate in the following steps. Such variations can be assigned to specific VOMs, as 3-me thylbutanal (wort), ethyl alcohol and ethyl octanoate (fermentation, maturation and filtration), or ethyl alcohol and isoamyl acetate (final product). These VOMs can influence the beer final flavour. Ethyl alcohol contributes to its strong and pungent smell and taste, while isoamyl acetate adds intense ‘fruity’ and ‘banana’ odours. These beer volatile fingerprints constitute a valuable tool to obtain insights on the impact of each brewing step on the final product, being also very useful for certification purposes.
- Extracellular volatilomic alterations induced by hypoxia in breast cancer cellsPublication . Taware, Ravindra; Taunk, Khushman; Kumar, Totakura V. S.; Pereira, Jorge A. M.; Câmara, José S.; Nagarajaram, H. A.; Kundu, Gopal C.; Rapole, SrikanthIntroduction The metabolic shift induced by hypoxia in cancer cells has not been explored at volatilomic level so far. The volatile organic metabolites (VOMs) constitute an important part of the metabolome and their investigation could provide us crucial aspects of hypoxia driven metabolic reconfguration in cancer cells. Objective To identify the altered volatilomic response induced by hypoxia in metastatic/aggressive breast cancer (BC) cells. Methods BC cells were cultured under normoxic and hypoxic conditions and VOMs were extracted using HS-SPME approach and profled by standard GC–MS system. Univariate and multivariate statistical approaches (p<0.05, Log2 FC≥0.58/≤−0.58, PC1>0.13/<−0.13) were applied to select the VOMs diferentially altered after hypoxic treatment. Metabolic pathway analysis was also carried out in order to identify altered metabolic pathways induced by the hypoxia in the selected BC cells. Results Overall, 20 VOMs were found to be signifcantly altered (p<0.05, PC1>0.13/< −0.13) upon hypoxic exposure to BC cells. Further, cell line specifc volatilomic alterations were extracted by comparative metabolic analysis of aggressive (MDA-MB-231) vs. non-aggressive (MCF-7) cells incubated under hypoxia and normoxia. In this case, 15 and 12 VOMs each were found to be signifcantly altered in aggressive cells when exposed to hypoxic and normoxic condition respectively. Out of these, 9 VOMs were found to be uniquely associated with hypoxia, 6 were specifc to normoxia and 6 were found common to both the conditions. Formic acid was identifed as the most prominent molecule with higher abundance levels in aggressive as compared to non-aggressive cells in both conditions. Furthermore, metabolic pathway analyses revealed that fatty acid biosynthesis and nicotinate and nicotinamide metabolism were signifcantly altered in aggressive as compared to non-aggressive cells in normoxia and hypoxia respectively. Conclusions Higher formate overfow was observed in aggressive cells compared to non-aggressive cells incubated under both the conditions, reinforcing its correlation with aggressive and invasive cancer type. Moreover, under hypoxia, aggres sive cells preferred to be bioenergetically more efcient whereas, under normoxia, fatty acid biosynthesis was favoured when compared to non-aggressive cells.
- Ultrasound-assisted liquid-liquid extraction followed by ultrahigh pressure liquid chromatography for the quantification of major carotenoids in tomatoPublication . Figueira, José A.; Pereira, Jorge A.M.; Porto-Figueira, Priscilla; Câmara, José S.Lycopene and β-carotene, the main carotenoids present in different tomatoes varieties (gordal, cherry, roma and campari) of Solanum lycopersicum L. (Solanaceae), were investigated using ultrasound-assisted liquid-liquid extraction (LLUSAE) followed by ultra-high pressure liquid chromatography with PDA detection (UHPLC-PDA) analysis. Experimental parameters influencing the LLUSAE were optimized using an univariate design, resulting in a 30-min ACN/MeOH extraction by sonication of a lyophilized sample, followed by PSA/C18/MgSO4 clean-up and fast centrifugation before UPLC analysis. Using this greener methodology, high recoveries (above 97%), good linearity (r2 > 0.98) and improved sensitivity, with limits of detection and quantification of 24.0 and 80.0 ng/mL for lycopene and 3.0 and 9.9 ng/mL for β-carotene, respectively, were obtained. This sensitivity is about five times better than previously reported in literature, making LLUSAE/UHPLC-PDA a promising strategy for lycopene and β-carotene quantification in tomato and eventually in other matrices. The carotenoids studied, lycopene and β-carotene, were found at highest concentrations in the gordal tomato variety, followed by cherry, roma and campari (727.1, 342.2, 267.2 and 218.2 μg/g and 80.4, 44.0, 45.7 and 44.0 μg/g for lycopene and β-carotene, respectively). Additionally, an exponential increase of both carotenoids occurs during ripening and mainly in the skin and locular cavity of the gordal variety. These results provide further evidences of the potential of tomatoes as an interesting source of lycopene and β-carotene.
- Microextraction using packed sorbent as an effective and high-throughput sample extraction technique: recent applications and future trendsPublication . Pereira, J.; Gonçalves, J.; Alves, V.; Câmara, J. S.In the last decade, several developments, particularly in the liquid chromatography and hyphenated techniques fields, have allowed researchers to reach analytical limits that are becoming very close to the single molecule level. Sample preparation, however, despite being a key step in the analytical methodology, did not track these developments and very few approaches able to cope with these stringent analytical requirements were developed. One such approach is microextraction by packed sorbent (MEPS), a sophisticated and miniaturized form of solid phase extraction that has reduced to the microliter scale every step of the sample extraction methodology including sample volume and solvent usage. Simultaneously, the amount of extraction phase used was also reduced and, more importantly, the sorbent is reusable dozens of times which significantly lowers the cost of analysis versus other solid-phase extraction approaches. In this review, we will update the state-of-the-art of the MEPS technique, focusing on the trends and applications reported since 2010 and future perspectives and developments that in our view will further improve the high-throughput potential and applications of this sample preparation methodology.
- Unveiling the bioactive potential of fresh fruit and vegetable waste in human health from a consumer perspectivePublication . Pereira, Jorge A. M.; Berenguer, Cristina V.; Andrade, Carolina F. P.; Câmara, José S.Food supply disruption and shortage verified during the current pandemic events are a scenario that many anticipate for the near future. The impact of climate changes on food production, the continuous decrease in arable land, and the exponential growth of the human population are important drivers for this problem. In this context, adding value to food waste is an obvious strategy to mitigate food shortages, but there is a long way to go in this field. Globally, it is estimated that one-third of all food produced is lost. This is certainly due to many different factors, but the lack of awareness of the consumer about the nutritional value of certain foods parts, namely peels and seeds, is certainly among them. In this review, we will unveil the nutritional and bioactive value of the waste discarded from the most important fresh fruit and vegetables consumed worldwide as a strategy to decrease food waste. This will span the characterization of the bioactive composition of selected waste from fruits and vegetables, particularly their seeds and peels, and their possible uses, whether in our diet or recycled to other ends.
- A micro-extraction technique using a new digitally controlled syringe combined with UHPLC for assessment of urinary biomarkers of oxidatively damaged DNAPublication . Mendes, Berta Rodrigues; Silva, Pedro; Aveiro, Fernando; Pereira, Jorge; Câmara, José S.The formation of reactive oxygen species (ROS) within cells causes damage to biomolecules, including membrane lipids, DNA, proteins and sugars. An important type of oxidative damage is DNA base hydroxylation which leads to the formation of 8-oxo-7,8-dihydro-29-deoxyguanosine (8-oxodG) and 5-hydroxymethyluracil (5-HMUra). Measurement of these biomarkers in urine is challenging, due to the low levels of the analytes and the matrix complexity. In order to simultaneously quantify 8-oxodG and 5-HMUra in human urine, a new, reliable and powerful strategy was optimised and validated. It is based on a semi-automatic microextraction by packed sorbent (MEPS) technique, using a new digitally controlled syringe (eVolH), to enhance the extraction efficiency of the target metabolites, followed by a fast and sensitive ultrahigh pressure liquid chromatography (UHPLC). The optimal methodological conditions involve loading of 250 mL urine sample (1:10 dilution) through a C8 sorbent in a MEPS syringe placed in the semi-automatic eVolH syringe followed by elution using 90 mL of 20% methanol in 0.01% formic acid solution. The obtained extract is directly analysed in the UHPLC system using a binary mobile phase composed of aqueous 0.1% formic acid and methanol in the isocratic elution mode (3.5 min total analysis time). The method was validated in terms of selectivity, linearity, limit of detection (LOD), limit of quantification (LOQ), extraction yield, accuracy, precision and matrix effect. Satisfactory results were obtained in terms of linearity (r2 . 0.991) within the established concentration range. The LOD varied from 0.00005 to 0.04 mg mL21 and the LOQ from 0.00023 to 0.13 mg mL21. The extraction yields were between 80.1 and 82.2 %, while inter-day precision (n=3 days) varied between 4.9 and 7.7 % and intra-day precision between 1.0 and 8.3 %. This approach presents as main advantages the ability to easily collect and store urine samples for further processing and the high sensitivity, reproducibility, and robustness of eVolHMEPS combined with UHPLC analysis, thus retrieving a fast and reliable assessment of oxidatively damaged DNA.
- Volatilomic insight of head and neck cancer via the effects observed on saliva metabolitesPublication . Taware, Ravindra; Taunk, Khushman; Pereira, Jorge A. M.; Shirolkar, Amey; Soneji, Dharmesh; Câmara, José S.; Nagarajaram, H. A.; Rapole, SrikanthHead and neck cancer (HNC) is a heterogeneous malignant disease with distinct global distribution. Metabolic adaptations of HNC are significantly gaining clinical interests nowadays. Here, we investigated effects of HNC on differential expression of volatile metabolites in human saliva. We applied headspace solid phase microextraction coupled with gas chromatography-mass spectrometry analysis of saliva samples collected from 59 human subjects (HNC - 32, Control - 27). We identified and quantified 48 volatile organic metabolites (VOMs) and observed profound effects of HNC on these metabolites. These effects were VOM specific and significantly differed in the biologically comparable healthy controls. HNC induced changes in salivary VOM composition were well attributed to in vivo metabolic effects. A panel of 15 VOMs with variable importance in projection (VIP) score >1, false discovery rate (FDR) corrected p-value < 0.05 and log2 fold change (log2 FC) value of ≥0.58/≤-0.58 were regarded as discriminatory metabolites of pathophysiological importance. Afterwards, receiver operator characteristic curve (ROC) projected certain VOMs viz., 1,4-dichlorobenzene, 1,2-decanediol, 2,5-bis1,1-dimethylethylphenol and E-3-decen-2-ol with profound metabolic effects of HNC and highest class segregation potential. Moreover, metabolic pathways analysis portrayed several dysregulated pathways in HNC, which enhanced our basic understanding on salivary VOM changes. Our observations could redefine several known/already investigated systemic phenomenons (e.g. biochemical pathways). These findings will inspire further research in this direction and may open unconventional avenues for non-invasive monitoring of HNC and its therapy in the future.
- The potential of microextraction techniques for the analysis of bioactive compounds in foodPublication . Pereira, Jorge A. M.; Casado, Natalia; Porto-Figueira, Priscilla; Câmara, José S.For a long time, the importance of sample preparation and extraction in the analytical performance of the most diverse methodologies have been neglected. Cumbersome techniques, involving high sample and solvent volumes have been gradually miniaturized from solid-phase and liquid-liquid extractions formats and microextractions approaches are becoming the standard in different fields of research. In this context, this review is devoted to the analysis of bioactive compounds in foods using different microextraction approaches reported in the literature since 2015. But microextraction also represents an opportunity to mitigate the environmental impact of organic solvents usage, as well as lab equipment. For this reason, in the recent literature, phenolics and alkaloids extraction from fruits, medicinal herbs, juices, and coffee using different miniaturized formats of solid-phase extraction and liquid-liquid microextraction are the most popular applications. However, more ambitious analytical limits are continuously being reported and emergent sorbents based on carbon nanotubes and magnetic nanoparticles will certainly contribute to this trend. Additionally, ionic liquids and deep eutectic solvents constitute already the most recent forefront of innovation, substituting organic solvents and further improving the current microextraction approaches.
- A fast method using a new hydrophilic–lipophilic balanced sorbent in combination with ultra-high performance liquid chromatography for quantification of significant bioactive metabolites in winesPublication . Silva, Catarina L.; Pereira, Jorge; Wouter, Van G.; Giró, Carme; Câmara, José S.This manuscript describes the development and validation of an ultra-fast, efficient, and high throughput analytical method based on ultra-high performance liquid chromatography (UHPLC) equipped with a photodiode array (PDA) detection system, for the simultaneous analysis of fifteen bioactive metabolites: gallic acid, protocatechuic acid, (−)-catechin, gentisic acid, (−)-epicatechin, syringic acid, p-coumaric acid, ferulic acid, m-coumaric acid, rutin, trans-resveratrol, myricetin, quercetin, cinnamic acid and kaempferol, in wines. A 50-mm column packed with 1.7-μm particles operating at elevated pressure (UHPLC strategy) was selected to attain ultra-fast analysis and highly efficient separations. In order to reduce the complexity of wine extract and improve the recovery efficiency, a reverse-phase solid-phase extraction (SPE) procedure using as sorbent a new macroporous copolymer made from a balanced ratio of two monomers, the lipophilic divinylbenzene and the hydrophilic N-vinylpyrrolidone (Oasis™ HLB), was performed prior to UHPLC–PDA analysis. The calibration curves of bioactive metabolites showed good linearity within the established range. Limits of detection (LOD) and quantification (LOQ) ranged from 0.006 μg mL−1 to 0.58 μg mL−1, and from 0.019 μg mL−1 to 1.94 μg mL−1, for gallic and gentisic acids, respectively. The average recoveries ± SD for the three levels of concentration tested (n = 9) in red and white wines were, respectively, 89 ± 3% and 90 ± 2%. The repeatability expressed as relative standard deviation (RSD) was below 10% for all the metabolites assayed. The validated method was then applied to red and white wines from different geographical origins (Azores, Canary and Madeira Islands). The most abundant component in the analysed red wines was (−)-epicatechin followed by (−)-catechin and rutin, whereas in white wines syringic and p-coumaric acids were found the major phenolic metabolites. The method was completely validated, providing a sensitive analysis for bioactive phenolic metabolites detection and showing satisfactory data for all the parameters tested. Moreover, was revealed as an ultra-fast approach allowing the separation of the fifteen bioactive metabolites investigated with high resolution power within 5 min.