Quercus spp. extract as a promising preventive or therapeutic strategy for cancer: A systematic review.

Acorns have traditionally been used in the human diet and for the treatment of specific diseases. Therefore, the present study performed a systematic review of studies which investigated the effects of Quercus spp. extracts in cancer prevention and treatment. A systematic literature search was performed for original records which addressed the anticancer effects of Quercus spp. extract in in vitro and in vivo cancer models. Body composition, food consumption, tumor development and/or toxicity were evaluated in in vivo studies, while cytotoxicity was evaluated in in vitro studies. Few studies and low sample sizes presented a challenge in the drawing of solid conclusions. Overall, the results suggested a positive impact of Quercus spp. extract, by reducing cancer development. Therefore, more studies with different cancer cell lines and animal models to address the efficacy of the acorn extracts in several types of cancer are required. Furthermore, the effects of acorn flour, incorporated in the diet, in an animal model of mammary cancer should be evaluated.

Exploring the Benefits of Phycocyanin: From Spirulina Cultivation to Its Widespread Applications.

Large-scale production of microalgae and their bioactive compounds has steadily increased in response to global demand for natural compounds. Spirulina, in particular, has been used due to its high nutritional value, especially its high protein content. Promising biological functions have been associated with Spirulina extracts, mainly related to its high value added blue pigment, phycocyanin. Phycocyanin is used in several industries such as food, cosmetics, and pharmaceuticals, which increases its market value. Due to the worldwide interest and the need to replace synthetic compounds with natural ones, efforts have been made to optimize large-scale production processes and maintain phycocyanin stability, which is a highly unstable protein. The aim of this review is to update the scientific knowledge on phycocyanin applications and to describe the reported production, extraction, and purification methods, including the main physical and chemical parameters that may affect the purity, recovery, and stability of phycocyanin. By implementing different techniques such as complete cell disruption, extraction at temperatures below 45 °C and a pH of 5.5-6.0, purification through ammonium sulfate, and filtration and chromatography, both the purity and stability of phycocyanin have been significantly improved. Moreover, the use of saccharides, crosslinkers, or natural polymers as preservatives has contributed to the increased market value of phycocyanin.

Study on the antineoplastic and toxicological effects of pomegranate (Punica granatum L.) leaf infusion using the K14-HPV16 transgenic mouse model.

Punica granatum L. (pomegranate) has been used in functional foods due to its various health benefits. However, the in vivo biological potential of its leaf remains little known. This study has aimed to characterize the antineoplastic and toxicological properties of using pomegranate leaf infusion (PLI) on transgenic mice carrying human papillomavirus (HPV) type 16 oncogenes. Thirty-eight mice were divided into 3 wild-type (WT) and 3 transgenic (HPV) groups, with exposure to 0.5% PLI, 1.0% PLI, and water. The animals' body weight, drink and food consumption were recorded. Internal organs, skin samples and intracardiac blood were collected to evaluate toxicological parameters, neoplastic lesions and oxidative stress. The results indicated that PLI was safe as no mortality, no behavioural disorders and no significant differences in the levels of microhematocrit, serum biochemical markers, internal organ histology, and oxidative stress was found among the WT groups. Histological analysis revealed that HPV animals that consumed PLI exhibited reduced hepatic, renal and cutaneous lesions compared with the HPV control group. Low-dose PLI consumption significantly diminished renal hydronephrosis lesions and relieved dysplasia and carcinoma lesions in the chest skin. Oxidative stress analysis showed that low-dose PLI consumption may have more benefits than high-dose PLI. These results suggest that oral administration of PLI has the potential to alleviate non-neoplastic and neoplastic lesions against HPV16-induced organ and skin injuries, though this requires further scientific research studies.

Acorn flour from holm oak (Quercus rotundifolia): Assessment of nutritional, phenolic, and technological profile.

Acorn is the fruit of holm oak (Quercus rotundifolia), being mainly used nowadays to feed animals, however a substantial part remains in the fields without any valorization. Underexploited crops are gaining new interest, driven by food security concerns and health benefits potential as well. In the present work, it was studied the physicochemical characteristics and functional perspective of acorn flour, as an ingredient for human diet. The study included nutritional composition analysis, phenolic compounds profile through HPLC, starch content and its microstructure, fibre, and pasting properties assessment. Acorn flour presented a high content in fat, particularly monounsaturated and polyunsaturated (oleic and linoleic acids), and high minerals content in particular K. Concerning phenolic profile, rutin, catechin, ellagic acid, gallic acid, and syringic acid were identified. In regards to technological profile, fibre was mainly insoluble, with around 11%, and starch content was 50%. Its pasting behaviour revealed a high gelatinization temperature (85 °C), with low breakdown, and higher retrogradation consistency. These results show acorn flour potential as a valuable and sustainable multipurpose food ingredient.

Food By-Product Valorization by Using Plant-Based Coagulants Combined with AOPs for Agro-Industrial Wastewater Treatment.

Re-using and adding value to by-products is one of the current focuses of the agri-food industry, following the Sustainable Development Goals of United Nations. In this work, the by-products of four plants, namely chestnut burr, acorn peel, olive leaf, and grape stem were used as coagulants to treat elderberry wastewater (EW), a problematic liquid effluent. EW pre-treatment using these natural coagulants showed promising results after pH and coagulant dosage optimization. However, the decrease in total organic carbon (TOC) was not significant, due to the addition of the plant-based natural coagulants which contain carbon content. After this pre-treatment, the photo-Fenton advanced oxidation process was selected, after preliminary assays, to improve the global performance of the EW treatment. Photo-Fenton was also optimized for the parameters of pH, H2O2, Fe2+, and irradiance power, and the best conditions were applied to the EW treatment. Under the best operational conditions defined in the parametric study, the combined results of coagulation-flocculation-decantation (CFD) and photo-Fenton for chestnut burr, acorn peel, olive leaf, and grape stem were, respectively, 90.2, 89.5, 91.5, and 88.7% for TOC removal; 88.7, 82.0, 90.2 and 93.1%, respectively, for turbidity removal; and finally, 40.6, 42.2, 45.3, and 39.1%, respectively, for TSS removal. As a final remark, it is possible to suggest that plant-based coagulants, combined with photo-Fenton, can be a promising strategy for EW treatment that simultaneously enables valorization by adding value back to food by-products.

Phytochemical and antioxidant analysis of medicinal and food plants towards bioactive food and pharmaceutical resources.

Plants with medicinal properties play an increasingly important role in food and pharmaceutical industries for their functions on disease prevention and treatment. This study characterizes the phenolic composition and antioxidant activity of seven medicinal and food plants, including the leaves of Salvia officinalis L., Rosmarinus officinalis L., Olea europaea L., and Punica granatum L., as well as the leaves and young stems of Ruta graveolens L., Mentha piperita L., and Petroselinum crispum, Mill., by using colorimetric, chromatographic, and spectrophotometric assays. Results revealed that the hydro-methanolic leaf extracts of P. granatum (pomegranate) displayed the highest content of total phenols (199.26 mg gallic acid per gram of plant dry weight), ortho-diphenols (391.76 mg gallic acid per gram of plant dry weight), and tannins (99.20 mg epicatechin per gram of plant dry weight), besides a higher content of flavonoids (24 mg catechin per gram of plant dry weight). The highest antioxidant capacity measured by ABTS, DPPH, and FRAP (2.14, 2.27, and 2.33 mM Trolox per gram of plant dry weight, respectively) methods was also obtained in pomegranate leaf extracts, being 4-200 times higher than the other species. Such potent antioxidant activity of pomegranate leaves can be ascribed to the presence of different types of phenolic compounds and the high content in tannins, whilst phenolic acids and flavonoids were found to be the dominant phenolic classes of the other six plants. Consequently, despite the well-known antioxidant properties of these plant species, our study suggests pomegranate leaf can stand out as a relatively more valuable plant source of natural bioactive molecules for developing novel functional food-pharma ingredients, with potential for not only promoting human health but also improving bio-valorization and environment.

Acorn Flour as a Source of Bioactive Compounds in Gluten-Free Bread.

Polyphenols are important bioactive compounds whose regular ingestion has shown different positive impacts in health. Celiac patients have nutritional deficiencies, bringing many problems to their health. Thus, it is important to develop gluten-free (GF) products, such as bread, with nutritional benefits. The acorn is the fruit of holm oak and cork oak, being an underexploited resource nowadays. Its nutritional and functional characteristics are remarkable: rich in unsaturated fatty acids and fiber, vitamin E, chlorophylls, carotenoids, phenolic compounds, and antioxidant properties. The purpose of this study was to assess the use of acorn flour as a bioactive compounds source and natural GF ingredient for baking GF bread. Bread loaves were prepared with buckwheat, rice, acorn flour, and potato starch. Two levels of acorn flour (23% and 35% of the flour mixture) were tested. The physical, nutritional, and sensory characteristics of the bread were analysed, as well as the composition of phenolic compounds: total phenols, ortho-diphenols, and flavonoids. The phenolic profile was assessed by Reverse Phase-High-Performance Liquid Chromatography-Diode Array Detector (RP-HPLC-DAD). The antioxidant activity of the bread extracts was determined by 2,2-azino-bis (3-ethylbenzothiazoline-6-sulphonic acid) diammonium salt (ABTS), diphenyl-1-picrylhidrazyl radical (DPPH), and ferric reducing antioxidant power (FRAP) methodologies. Acorn flour can be considered a good source of bioactive compounds and antioxidants in GF bread. Acorn flour showed good technological properties in GF baking, improving bread nutritional and sensory characteristics.

Comparison of near-infrared (NIR) and mid-infrared (MIR) spectroscopy for the determination of nutritional and antinutritional parameters in common beans.

Common beans (Phaseolus vulgaris L.), represent the most consumed legume worldwide and constitute an important source of protein, being also known to contain antinutritional compounds, which compromise nutrients' bioavailability. However, the standard methodologies to assess these constituents are time-consuming and complex. Therefore, the present study evaluated the suitability of near-infrared (NIR) and mid-infrared (MIR) spectroscopies for the development of simple and reliable methods to assess protein, lipids, tannins and phytic acid contents, besides specific amino acids, in whole bean flours. Partial least squares (PLS) regression was used to develop analytical models, and external validation was performed. NIR displayed better performance for the evaluation of protein, lipids, tannins and phytic acid contents, and MIR, for the assessment of specific amino acids. In both techniques, the use of the 1st derivative was the best data treatment. Overall, both techniques represent reliable methods to evaluate the proximate and antinutritional composition of bean flours.

Critical Review on the Significance of Olive Phytochemicals in Plant Physiology and Human Health.

Olive oil displays remarkable organoleptic and nutritional features, which turn it into a foodstuff appreciated by consumers, and a basic component of the Mediterranean diet. Indeed, the noticed benefits of including olive oil in the diet have been assigned to the presence of diverse bioactive compounds with different molecular structures. These compounds confer a wide range of biological properties to this food matrix, including the prevention of distinct human diseases as well as the modulation of their severity. The most relevant bioactive compounds present in olive oil correspond to benzoic and cinnamic acids, phenolic alcohols and secoiridoids, and also flavonoids. Over the last decades, several studies, devoted to gaining a further insight into the relative contribution of the separate groups and individual compounds for their biological activities, have been conducted, providing relevant information on structure-activity relationships. Therefore, this paper critically reviews the health benefits evidenced by distinct phenolic compounds found in olive oils, thus contributing to clarify the relationship between their chemical structures and biological functions, further supporting their interest as essential ingredients of wholesome foods.

Valorization Challenges to Almond Residues: Phytochemical Composition and Functional Application.

Almond is characterized by its high nutritional value; although information reported so far mainly concerns edible kernel. Even though the nutritional and commercial relevance of the almond is restricted to almond meat; to date; increasing attention has been paid to other parts of this fruit (skin; shell; and hull); considered by-products that are scarcely characterized and exploited regarding their properties as valuable sources of bioactive compounds (mainly represented by phenolic acids and flavonoids). This lack of proper valorization procedures entails the continuation of the application of traditional procedures to almond residues that nowadays are mainly addressed to livestock feed and energy production. In this sense; data available on the physicochemical and phytochemical composition of almond meat and its related residues suggest promising applications; and allow one to envisage new uses as functional ingredients towards value-added foods and feeds; as well as a source of bioactive phytochemicals to be included in cosmetic formulations. This objective has prompted investigators working in the field to evaluate their functional properties and biological activity. This approach has provided interesting information concerning the capacity of polyphenolic extracts of almond by-products to prevent degenerative diseases linked to oxidative stress and inflammation in human tissues and cells; in the frame of diverse pathophysiological situations. Hence; this review deals with gathering data available in the scientific literature on the phytochemical composition and bioactivity of almond by-products as well as on their bioactivity so as to promote their functional application.

New grape stems' isolated phenolic compounds modulate reactive oxygen species, glutathione, and lipid peroxidation in vitro: Combined formulations with vitamins C and E.

The antioxidant potential of grape (Vitis vinífera L.) stems has been reported in the last decade although no identification of the individual compounds responsible for such action has been done. In this work, polyphenolic extract of grape stems was processed resorting to semi-preparative HPLC, allowing to obtain 5 purified polyphenols (caftaric acid, malvidin-3-O-glucoside, quercetin-3-O-glucuronide, mailvidin-3-O-(6-O-caffeoyl)-glucoside, and Σ-viniferin), which were fully characterized by HPLC-PDA-ESI-MSn. Isolated compounds were featured on their radical scavenging capacity (DPPH and ABTS), cell viability, anti-inflammatory activity, and capacity to modulate the level of reactive oxygen species, glutathione, lipid peroxidation, and overall oxidative stress in a biological model (human keratinocytes) in vitro, under basal and oxidative conditions. The results obtained noticed the combinations malvidin-3-O-glucoside+Vitamin E and quercetin-3-O-glucuronide+vitamin C as the most effective, allowing to improve the capacity of complete extracts or individual compounds, and being candidates to be used in the development of new functional products.

Oxidative stress prevention and anti-apoptosis activity of grape (Vitis vinifera L.) stems in human keratinocytes.

To date, grape stems have been partially assessed on their content in phenolics and their radical scavenging activity, whilst the potential to modulate oxidative stress in biological models remains underexplored. In the present work, the effect of grape stems' phenolics on redox unbalance was evaluated in human keratinocytes (HaCaT cells). Grape stems' extracts were assessed on their phenolic composition by high performance liquid chromatography coupled with photodiode array detection and electrospray ionization-mass spectrometry (HPLC-PAD-ESi-MSn), besides on radical scavenging capacity (ABTS and DPPH). In addition, their protective effect against oxidative stress induced by H2O2 by the determination of the level of glutathione, reactive oxygen species, lipid peroxidation, and overall oxidative stress in HaCaT cells by flow cytometry was evaluated. This characterization allowed to identify five flavonols, one cinnamic acid, and one stilbene. A close correlation between the concentration of these phenolics and the capacity to scavenge free radicals and with the potential to modulate the redox balance in vitro was observed. From the analysis of correlation, the activity of malvidin-3-O-glucoside, malvidin-3-O-(6-O-caffeoyl)-glucoside, and malvidin-3-O-rutinoside with respect to the prevention of basal oxidative stress and the capacity of isorhamnetin-3-O-(6-O-feruloyl)-glucoside and kaempferol-3-O-rutinoside to prevent H2O2-induced redox unbalance were stated. Furthermore, grape stems' phenolics also showed an efficient capacity to modulate apoptosis in HaCaT cells, reducing the frequency of annexin V/PI double positive apoptotic cells by up to 99.5% relative to controls, which was further confirmed by the determination of the appearance of the occurrence of apoptotic bodies and the expression of activated (cleaved) caspase-3 by flow cytometry and western-blot, respectively. These results supported the potential of individual phenolics from grape stems to modulate oxidative stress, allowing to envisage dedicated combinations of single compounds for the development of efficient formulations efficient against oxidative stress.

Trace Element Content of Monovarietal and Commercial Portuguese Olive Oils.

Olive (Olea europaea L.) represents one of the most important and ancient crops in the Mediterranean countries, being widely known due to the optimal organoleptic characteristics of its oil, besides the important health benefits assigned to this product in the last few years. Since olive oil composition, respecting specific trace elements, has a direct impact on the quality and safety of this product, eleven monovarietal and twelve commercial olive oils were analysed spectrometrically concerning their metal composition. Regarding quality parameters, the monovarietal olive oils display larger quantities of Mg, Fe, Mn, while the commercial samples are characterized by greater amounts of Na, Ca, Cu and Zn. In which respects to safety parameters, the commercial samples present higher values for Al and Pb, and lower quantities of Cd, respecting the monovarietal ones. The assessment of the values observed for the trace elements, resorting to multivariate statistical methods, allowed to clearly distinguish the type of olive oil production-monovarietal or commercial blend. For this purpose, besides Cluster analysis, the data have been subjected to Principal Component Analysis-undertaken for either all the evaluated parameters simultaneously, or the quality/safety parameters separately, both data sets allowing the discrimination of the samples.

Short wavelength Raman spectroscopy applied to the discrimination and characterization of three cultivars of extra virgin olive oils in different maturation stages.

Extra virgin olive oils produced from three cultivars on different maturation stages were characterized using Raman spectroscopy. Chemometric methods (principal component analysis, discriminant analysis, principal component regression and partial least squares regression) applied to Raman spectral data were utilized to evaluate and quantify the statistical differences between cultivars and their ripening process. The models for predicting the peroxide value and free acidity of olive oils showed good calibration and prediction values and presented high coefficients of determination (>0.933). Both the R(2), and the correlation equations between the measured chemical parameters, and the values predicted by each approach are presented; these comprehend both PCR and PLS, used to assess SNV normalized Raman data, as well as first and second derivative of the spectra. This study demonstrates that a combination of Raman spectroscopy with multivariate analysis methods can be useful to predict rapidly olive oil chemical characteristics during the maturation process.

Discrimination and characterisation of extra virgin olive oils from three cultivars in different maturation stages using Fourier transform infrared spectroscopy in tandem with chemometrics.

A methodology based on Fourier transform infrared (FTIR) spectroscopy, combined with multivariate analysis methods, was applied in order to monitor extra virgin olive oils produced from three distinct cultivars on different maturation stages. For the first time, this kind of methodology is used for the simultaneous discrimination of the maturation stage, and different cultivars. Principal component analysis and discriminant analysis were utilised to create a model for the discrimination of olive oil samples. Partial least squares regression was employed to design calibration models for the determination of chemical parameters. The performance of these models was based on the multiple coefficient of determination (R(2)), the root mean square error of calibration (RMSEC) and root mean square error of cross validation (RMSECV). The prediction models for the chemical parameters resulted in a R(2) ranged from 0.93 to 0.99, a RMSEC ranged from 1% to 4% and a RMSECV from 2% to 5%. It has been shown that this kind of approach allows to distinguish the different cultivars, and to clearly discern the different maturation stages, in each one of these distinct cultivars. Furthermore, the results demonstrated that FTIR spectroscopy in tandem with chemometric techniques allows the creation of viable and accurate models, suitable for correlating the data collected by FTIR spectroscopy, with the chemical composition of the EVOOs, obtained by standard methods.

A novel, direct, reagent-free method for the detection of beeswax adulteration by single-reflection attenuated total reflectance mid-infrared spectroscopy.

In this work, a novel, direct, reagent-free method for the detection of beeswax adulteration by paraffin, microcrystalline wax, tallow and stearic acid using single-reflection attenuated total reflectance mid-infrared spectroscopy was developed. The use of the absorbance ratios of [Formula: see text] , [Formula: see text] and [Formula: see text] allows a minimum of 5% paraffin/microcrystalline wax and tallow adulteration and 0.5% stearic acid adulteration of beeswax to be detected. The upper and lower critical limits for beeswax authenticity were established from the analysis of virgin beeswax and were validated by independent analysis of real sheet and comb beeswax samples using high-temperature gas chromatography with flame-ionization detection. In addition to its simplicity with respect to sample handling, the amount of sample and the time needed are far less than those required in previously described methods, which are based on chemical analysis and chromatographic techniques. These advantages result in time and cost savings, an increase in the number of samples that can be analyzed, and, most importantly, the detection of the main beeswax adulterants using a single method.

Oxidation of mannosyl oligosaccharides by hydroxyl radicals as assessed by electrospray mass spectrometry.

The hydroxyl radicals are widely implicated in oxidation of carbohydrates during biological and industrial processes being responsible for their structural modifications and causing functional damage. The identification of intermediate oxidation products is hampered by a lack of reliable sensible methods for their detection. In this study, the oxidation of two models of galactomannans (Man(3) and GalMan(2)) has been studied in reaction with hydroxyl radical generated by Fenton reaction. The oxidation patterns were assessed using preparative ligand-exchange/size-exclusion chromatography (LEX/SEC) coupled with tandem electrospray mass spectrometry (ESI-MS/MS). This allowed the identification of derived oligosaccharides (OS) containing hexuronic, hexonic, pentonic and erythronic acid residues and neutral OS bearing hydroperoxy, hydrated carbonyl moieties and residues from pyranosyl ring cleavage. The depolymerization products have been also detected upon oxidation of oligomers. This study allowed developing a simple, effective 'fingerprinting' protocol for detecting the damage done to mannans by oxidative radicals.

Effect of cooking on total vitamin C contents and antioxidant activity of sweet chestnuts (Castanea sativa Mill.).

In this work the total vitamin C contents (ascorbic acid+dehydroascorbic acid) and antioxidant activity of raw and cooked chestnuts was evaluated. The vitamin C contents of raw chestnuts varied significantly between the different cultivars (cv) studied and it varied from 400mg/kg dry weight (cv Lada) to 693mg/kg dry weight (cv Martaínha). The different cultivars behave differently during the cooking process concerning the loss of vitamin C. A significant decrease in the vitamin C content of the chestnuts was observed, 25-54% for the boiling process and 2-77% for the roasting process. Boiled and roasted chestnuts can be good sources of vitamin C since it may represent 22.4%, 16.2%, 26.8% and 19.4%, respectively, of the recommended dietary intake for an adult man and woman. The cooking process significantly changed the antioxidant activity of the chestnuts. A difference was observed between the cultivars during the cooking processes, concerning the antioxidant activity. For the raw chestnuts the variation in vitamin C content of the chestnuts explains 99% of the antioxidant activity variation but for the roasted and boiled chestnuts this percentage significantly decreases to 51% and 88%, respectively. Although a high antioxidant activity is still present in the cooked chestnuts, the cause for this antioxidant activity is less dependent on the vitamin C content of the chestnuts, probably due to the conversion of ascorbic acid to dehydroascorbic acid. The increase in gallic acid during the cooking process, presumably transferred from the peels to the fruit, also contributes to the high antioxidant activity observed for the cooked chestnuts.

A fast, simple, and reliable hydrophilic interaction liquid chromatography method for the determination of ascorbic and isoascorbic acids.

A reliable method for the determination of total vitamin C must be able to resolve ascorbic acid (AA) and the epimeric isoascorbic acid (IAA) and determine the sum of AA and its oxidized form dehydroascorbic acid. AA and IAA are polar molecules with a low retention time in conventional reversed phase systems, and hence of difficult resolution. Hydrophilic interaction chromatography using a TSKgel Amide-80 stationary phase with isocratic elution was successful in resolving the two epimers. The column was compatible with injections of high concentrations of metaphosphoric acid, tris(2-carboxyethyl)-phosphine, and EDTA without drift of baseline and retention time. Total AA and IAA were extracted, stabilized, and reduced in one step at 40 °C, using 5% m-phosphoric acid, 2 mM of EDTA, and 2 mM of tris(2-carboxyethyl)-phosphine as reducing agent. This simple, fast, and robust hydrophilic interaction chromatography-DAD method was applied for the analysis of food products namely fruit juices, chestnut, and ham and also in pharmaceutical and multivitamin tablets. Method validation was performed on the food products, including parameters of precision, accuracy, linearity, limit of detection, and quantification (LOQ). The absence of matrix interferences was assessed by the standard addition method and Youden calibration. The method was fast, accurate, and precise with a LOQ(AA) of 1.5 mg/L and LOQ(IAA) of 3.7 mg/L. The simple experimental procedure, completed in 1 h, the possibility of using IAA as an internal standard, and low probability of artifacts are the major advantages of the proposed method for the routine determination of these compounds in a large number of samples.

Structural characterization of nitrated 2'-hydroxychalcones by electrospray ionization tandem mass spectrometry.

Isomeric 2'-hydroxychalcones bearing nitro and methoxy groups in different positions of their skeleton were analyzed by tandem mass spectrometry (MS/MS) with electrospray ionization (ESI), in positive mode. Collision-induced dissociation of the protonated molecules, [M + H](+), formed under electrospray conditions were studied and it was found that the product ion spectra of these chalcones presented different fragmentation patterns depending on the position of the substituents on the molecule. The product ion spectra (ESI- MS/MS) of the B ring ortho-nitro substituted 2'-hydroxychalcone and of the 4'-methoxychalcones showed loss of OH, 2OH and combined losses of OH and H(2)O. These fragment ions were absent in the spectra of the respective meta- and para isomers. The observed differences in the product ion spectra of these nitrochalcones allowed identification of the o-nitro derivatives. Distinction between the meta- and para derivatives was not achieved. Chalcones bearing 6'-methoxy substituents showed distinct fragmentation from the one observed for their isomers, 4'-methoxychalcones, since they present only one fragment ion, a typical ((0,alpha)A - H)(+) and, therefore, do not allow detailed structural information to be obtained, nor to differentiate between the o-, m- or p-nitro isomers. Overall, it was found that small changes in the substitution pattern of chalcones change their fragmentation considerably in the ESI-MS/MS, and that these features permit the differentiation of specific isomers of these 2'-hydroxynitrochalcones.