Phenolic compounds and antioxidant capacity of three thymus species plants

Thymus plants comprise distinct species with claimed health properties [1], commonly associated to their essential oils and phenolic compounds. Albeit that, the phenolic composition and the biological activities of many Thymus species remain unclear. This work aimed to elucidate the phenolic composition and antioxidant properties of aqueous extracts from Thymus herba barona, Thymus caespetitus and Thymus fragrantissimus. The aqueous extracts of the three Thymus species were evaluated for their total phenolic compounds by an adaptation of the Folin-Ciocalteu method [2], and individual phenolic compounds were identified by high performance liquid chromatography associated with electrospray mass spectrometry (HPLC-DAD-ESI-MSn) in the negative mode. The antioxidant activity of each extract was carried out by DPPH● scavenging assay and ferric reducing antioxidant power assays [3]. Total phenolic compounds in the three extracts ranged from 236±27 (T. caespetitus) to 273±17 μg GAE/mg (T. fragrantissimus). Similarly to other Thymus species [1,4], these extracts were rich in caffeic acid derivatives (characteristic UV spectra maxima at 290 and 328 nm) and mainly composed of rosmarinic acid (MW 360). Other caffeic acid derivatives included salvianolic acid K (MW 556) and 3′-O-(8″-Z-caffeoyl)rosmarinic acid (MW 538). High amounts of the flavone luteolin-O-glucuronide ([M-H]− at m/z 461→285) were found in T. caespetitus while the others species contained moderate amounts of this compound. T. herba barona, T. caespetitus and T. fragrantissimus extracts showed high DPPH radical scavenge ability (EC50 values 11.6±0.9, 13.8±0.6 and 10.9±1.2 μg/mL respectively), as well as high reducing power (EC50 values of 35.1±4.5, 39.3±2.7 and 32.4±4.3 μg/mL, respectively), that were comparable to those of reference compounds. This work is an important contribution for the phytochemical characterization and the antioxidant capacity of these three Thymus species.

Phenolic compounds and antioxidant capacity of three thymus species plants

Thymus plants comprise distinct species with claimed health properties [1], commonly associated to their essential oils and phenolic compounds. Albeit that, the phenolic composition and the biological activities of many Thymus species remain unclear. This work aimed to elucidate the phenolic composition and antioxidant properties of aqueous extracts from Thymus herba barona, Thymus caespetitus and Thymus fragrantissimus. The aqueous extracts of the three Thymus species were evaluated for their total phenolic compounds by an adaptation of the Folin-Ciocalteu method [2], and individual phenolic compounds were identified by high performance liquid chromatography associated with electrospray mass spectrometry (HPLC-DAD-ESI-MSn) in the negative mode. The antioxidant activity of each extract was carried out by DPPH● scavenging assay and ferric reducing antioxidant power assays [3]. Total phenolic compounds in the three extracts ranged from 236±27 (T. caespetitus) to 273±17 μg GAE/mg (T. fragrantissimus). Similarly to other Thymus species [1,4], these extracts were rich in caffeic acid derivatives (characteristic UV spectra maxima at 290 and 328 nm) and mainly composed of rosmarinic acid (MW 360). Other caffeic acid derivatives included salvianolic acid K (MW 556) and 3′-O-(8″-Z-caffeoyl)rosmarinic acid (MW 538). High amounts of the flavone luteolin-O-glucuronide ([M-H]− at m/z 461→285) were found in T. caespetitus while the others species contained moderate amounts of this compound. T. herba barona, T. caespetitus and T. fragrantissimus extracts showed high DPPH radical scavenge ability (EC50 values 11.6±0.9, 13.8±0.6 and 10.9±1.2 μg/mL respectively), as well as high reducing power (EC50 values of 35.1±4.5, 39.3±2.7 and 32.4±4.3 μg/mL, respectively), that were comparable to those of reference compounds. This work is an important contribution for the phytochemical characterization and the antioxidant capacity of these three Thymus species.

Optimization of microwave-assisted extration of phenolic compounds from tomao by full factorial design coupled with response surface methodology

Tomato (Lycopersicon esculentum Mill.), apart from being a functional food rich in carotenoids, vitamins and minerals, is also an important source of phenolic compounds [1 ,2]. As antioxidants, these functional molecules play an important role in the prevention of human pathologies and have many applications in nutraceutical, pharmaceutical and cosmeceutical industries. Therefore, the recovery of added-value phenolic compounds from natural sources, such as tomato surplus or industrial by-products, is highly desirable. Herein, the microwave-assisted extraction of the main phenolic acids and flavonoids from tomato was optimized. A S-Ieve! full factorial Box-Behnken design was implemented and response surface methodology used for analysis. The extraction time (0-20 min), temperature (60-180 "C), ethanol percentage (0-100%), solidlliquid ratio (5-45 g/L) and microwave power (0-400 W) were studied as independent variables. The phenolic profile of the studied tomato variety was initially characterized by HPLC-DAD-ESIIMS [2]. Then, the effect of the different extraction conditions, as defined by the used experimental design, on the target compounds was monitored by HPLC-DAD, using their UV spectra and retention time for identification and a series of calibrations based on external standards for quantification. The proposed model was successfully implemented and statistically validated. The microwave power had no effect on the extraction process. Comparing with the optimal extraction conditions for flavonoids, which demanded a short processing time (2 min), a low temperature (60 "C) and solidlliquid ratio (5 g/L), and pure ethanol, phenolic acids required a longer processing time ( 4.38 min), a higher temperature (145.6 •c) and solidlliquid ratio (45 g/L), and water as extraction solvent. Additionally, the studied tomato variety was highlighted as a source of added-value phenolic acids and flavonoids.

Contribution of plant-derived phenolic compounds to combat Candida species biofilms

Opportunistic fungal infections, namely involving Candida species, constitute a hot topic for scientific researchers. The present wor1( aims to access antifungal potential of plant-derived phenolic extrac:ls against planktonic cells and biofilms of Candida species. Eucalyptvs globulus Labill. (blue gum), Glycyrrhiza glabra L. (licorice), Juglans regia L. (walnut) and Salvia officina/is L. (sage) evidenced to be the most effective Candida growth inhibitors, using disc diffusion assay. Minmal inhibitory (MIC) and minimal fungicidal (MFC) concentrations, and chemical composition of extracts by using HPLC-DAO-ESVMS were also determined. Blue gum and walnut mainly exerted fungistatic potential, while sage exerted an interesting anti-Candida potential. However, the most prominent candidacidal potential was observed to licorice extract, being achieved the lowest MIC and MFC values. The candidacidal potential of these phenolic extracts was mainly attributed to their high abundance in flavonoids, mainly flavones: luteolin (sage) and apigen~ derivatives (licorice), and flavanones: liQuiritin derivatives (licorice). In order to deepen the knowledge on the most effective extract. its abiity to inhibit biofilm formation was evaluated. Overall, a double concentration of MFC value was necessary to achieve similar results in biofims. Row cytometry assays were also carried out, and the obtained results revealed that primary lesion of cellular membrane appear to be most relevant mode of action. Thus, plant derived phenolic compounds evidence a promising potential to combat Candida species biofilms, both individually or combined with conventional therapy.

Analysis of phenolic compounds in Cynara scolymus L. and Silybum marianum (L.) Gaertn. by HPLC-DAD-ESI/MS

Cynara scolymus L. (artichoke) and Silybum marianum (L.) Gaertn. (milk thistle) are medicinal plants native to the Mediterranean Basin that belong to the Asteraceae family. The flowers and leaves of milk thistle are used in the treatment of liver, spleen and gallbladder disorders [1] and artichoke leaves are used for their cholagogue, choleretic and choliokinetic actions, and also for treatment of dyspepsia and as antidiabetics [2]. The beneficial properties of medicinal plants can be related to their large diversity of phytochemicals, among which phenolic compounds are outstanding. Thereby, the aim of the present work was to obtain and compare the phenolic profiles of artichoke and milk thistle aqueous (prepared by infusion) and hydromethanolic (maceration in methanol: water 80:20, v/v) extracts, using HPLC-DAD-ESI/MS. The aqueous extract of artichoke presented higher concentration in total phenolic compounds (15.29 mg/g extract) than the hydromethanolic extract (4.37 mg/g) with slight differences between the respective profiles; the major flavonoid found in the aqueous and hydromethanolic extract was luteolin-7-O-glucuronide (5.64 and 0.70 mg/g, respectively), followed by luteolin-7-O-glucoside (2.88 and 0.49 mg/g, respectively). Monocaffeoylquinic acid derivatives were only present in the hydromethanolic extract, being 5-O-caffeoylquinic acid (0.49 mg/g) the most abundant one, while dicaffeoylquinic acid derivatives were mostly identified in the aqueous extract; 1,3-O-dicaffeoylquinic acid was the most abundant one in both extracts (0.90 and 0.37 mg/g in the aqueous and hydromethanolic extract, respectively). Regarding to milk thistle preparations, similar phenolic profiles were observed, with only quantitative differences between them. The aqueous extract revealed a higher phenolic compounds concentration (5.57 mg/g) than the hydromethanolic extract (3.56 mg/g), with apigenin-7-O-glucuronide as the major compound in both preparations (3.14 mg/g in the aqueous extract, and 0.58 mg/g in the hydromethanolic extract). Total flavonoids were higher in the aqueous extract (4.66 mg/g), with apigenin-7-Oglucuronide, luteolin-7-O-glucuronide (1.17 mg/g), and apigenin-O-deoxyhexosylglucuronide (0.36 mg/g) as the main constituents. The phenolic acids found in the hydromethanolic extract (total content 1.65 mg/g), included 5-O-caffeolyquinic and protocatechuic acids (0.56 and 0.44 mg/g, respectively). Besides these phenolic acids, the hydromethanolic extract also revealed high levels of luteolin-7-O-glucuronide (0.58 mg/g). Overall, aqueous extracts presented higher phenolic contents than their hydromethanolic extracts in both species, which could be related with the heat treatment to which infusions were subjected.

Phenolic compounds in Coleostephus myconis (L.) Rchb.f.: characterization by HPLC-DAD-ESI/MS and phenology effects

The Asteraceae family is spread worldwide. In Portugal, there are more than 300 species, standing out as one of the botanical families with largest representation in the Portuguese flora. Coleostephus myconis (L.) Rchb.f. is a scarcely studied Asteraceae species, characterized as having ruderal growth and persistence in abandoned soils (an expanding problem due to the desertification phenomena in rural areas). In this work, the flowers of C. myconis were collected in three different flowering stages (i: flower bud; ii: flower in anthesis; iii: senescent flower) from the Northwestern area of the Portuguese territory. Powdered samples (1 g) were extracted twice with ethanol:water 50:50 (v/v). After removing solvents, the combined extracts were re-dissolved, filtered through 0.22-μm disposable LC filter disks and analyzed by high performance liquid chromatography coupled to a diode array detector and electrospray ionization-mass spectrometry (HPLC-DAD/ESI-MS). The phenolic compounds were characterized according to their UV and mass spectra, and retention times. For the quantitative analysis, calibration curves of standard compounds were used. According to the UV spectra (λmax = 314-330 nm) and pseudomolecular ions ([M-H]-) at m/z 353 and 515, all producing an m/z 191 ion, four compounds derived from quinic acid were detected: 3-O-caffeoylquinic acid (Figure 1A), 5-O-caffeoylquinic acid (Figure 1B), 3,5-O-dicaffeoylquinic acid (Figure 1C) and 4,5-O-dicaffeoylquinic acid (Figure 1D), as also supported by the literature [1,2]. A fifth phenolic acid was identified as protocatechuic acid. The detected flavonoid were quercetin-O-glucuronide, quercetin-3-Oglucoside, myricetin-O-methyl-hexoside and a second glycosylated myricetin (not possible to identify completely). Some statistically significant changes were detected among the different assayed flowering stages; nevertheless, 3,5-O-dicaffeoylquinic acid was the major compound, independently of the phenologic stage. According to the previous results, C. myconis might be considered as a potential natural source of these valuable bioactive compounds, especially considering the high botanical representativeness of this plant and its inexpensiveness.

HPLC-DAD-ESI/MS analysis of phenolic compounds in four tomato varieties and evaluation of the antioxidant capacity

Tomato (Lycopersicon esculentum L.) is the second most important vegetable crop worldwide and a key component in the so-called “Mediterranean diet”. In the Northeastern region of Portugal, local populations still prefer to consume traditional tomato varieties which they find very tasty and healthy, as they are grown using extensive farming techniques. A previous study of our research team described the nutritional value of the round (batateiro), long (comprido), heart (coração) and yellow (amarelo) tomato varieties [1], but the phenolic profile was unknown until now. Thus, the objective of this study was to characterize the phenolic profiles of these four tomato farmers’ varieties by using HPLC-DAD-ESI/MS and evaluate its antioxidant capacity through four in vitro assays based on different reaction mechanisms. A cis p-coumaric acid derivative was the most abundant compound in yellow and round tomato varieties, while 4-O-caffeolyquinic acid was the most abundant in long and heart varieties. The most abundant flavonoid was quercetin pentosylrutinoside in the four tomato varieties. Yellow tomato presented the highest levels of phenolic compounds, including phenolic acids and flavonoids, but the lowest antioxidant activity. In turn, the round tomato gave the best results in all the antioxidant activity assays. This study demonstrated that these tomato farmers’ varieties are a source of phenolic compounds, mainly phenolic acid derivatives [2], and possess high antioxidant capacity [1]; being thus key elements in the diet to prevent chronic degenerative diseases associated to oxidative stress, such as cancer and coronary artery disease.

Exploring plant tissue culture to improve the production of phenolic compounds: A review

Plant tissue and organ culture has been extensively used from the beginning of the XX century for the study and comprehension of some primary biological mechanisms such as morphogenesis. However, with the increasing demand of the market for novel products derived from plants, in vitro culture became a reliable technique for the mass production of plant material. Moreover, the potential to use this technique for the production of some bioactive compounds, such as phenolic compounds, is immense since it allows the manipulation of the biosynthetic routes to increase the production and accumulation of specific compounds. This work intends to make a brief historical review of in vitro culture, highlighting its use for the production of bioactive compounds. Also, emphasizes the importance of phenolic compounds for the consumer as well reviews the metabolic pathways involved in its production in plant cells. Furthermore, it was carried out a comprehensive study on the work developed for the production of plant phenolic compounds in in vitro cultures, as well as on the type of elicitors used to increase of the same production; also a brief highlighting of the phenolic compounds which serve as elicitors. There are numerous reports directed to the production of phenolic extracts in in vitro plant cultures, however there is a lack in the production of individual phenolic compounds mainly due to the complexity of the biosynthetic routes and extraction procedures. Elicitation procedures are often used to increase the production of phenolics, archieving in most cases higher yields than in non-elicitated cultures. The increasing production of bioactive phenolic extracts/compounds allows for their further applicability, namely in the industry of functional foods or in pharmaceutical/medical fields.

Wild Roman chamomile extracts and phenolic compounds: enzymatic assays and molecular modelling studies with VEGFR-2 tyrosine kinase

Angiogenesis is a process by which new blood vessels are formed from the pre-existing vasculature, and it is a key process that leads to tumour development. Some studies have recognized phenolic compounds as chemopreventive agents; flavonoids, in particular, seem to suppress the growth of tumor cells modifying the cell cycle. Herein, the antiangiogenic activity of Roman chamomile (Chamaemelum nobile L.) extracts (methanolic extract and infusion) and the main phenolic compounds present (apigenin, apigenin-7-O-glucoside, caffeic acid, chlorogenic acid, luteolin, and luteolin-7-O-glucoside) was evaluated through enzymatic assays using the tyrosine kinase intracellular domain of the Vascular Endothelium Growth Factor Receptor-2 (VEGFR-2), which is a transmembrane receptor expressed fundamentally in endothelial cells involved in angiogenesis, and molecular modelling studies. The methanolic extract showed a lower IC50 value (concentration that provided 50% of VEGFR-2 inhibition) than the infusion, 269 and 301 μg mL(-1), respectively. Regarding phenolic compounds, luteolin and apigenin showed the highest capacity to inhibit the phosphorylation of VEGFR-2, leading us to believe that these compounds are involved in the activity revealed by the methanolic extract.

In vivo antioxidant activity of phenolic compounds: facts and gaps

Numerous diseases have been related with free radicals overproduction and oxidative stress. Botanical preparations possess a multitude of bioactive properties, including antioxidant potential, which has been mainly related with the presence of phenolic compounds. However, the mechanisms of action of these phytochemicals, in vivo effects, bioavailability and bio-efficacy still need research. Scope and Approach: The present report aims to provide a critical review on the aspects related with the in vivo antioxidant activity of phenolic extracts and compounds from plant origin. Key findings: Biological functions beyond the human metabolism were discussed, comparing in vivo vs. in vitro studies, as also focusing the conditioning factors for phenolic compounds bioavailability and bio-efficacy. Furthermore, an upcoming perspective about the use of phytochemicals as life expectancy promoters and anti-aging factors in human individuals was provided. Conclusions: Overall, and despite all of those advances, the study of the biological potential of numerous natural matrices still remains a hot topic among the scientific community. In fact, the available knowledge about the responsible phytochemicals for the biological potential, their mechanisms of action, the establishment of therapeutic and prophylactic doses, and even the occurrence of biochemical inter-relations, is considerable scarce.

Extraction, identification, fractionation and isolation of phenolic compounds in plants with hepatoprotective effects

The liver is one of the most important organs of human body, being involved in several vital functions and regulation of physiological processes. Given its pivotal role in the excretion of waste metabolites and drugs detoxification, the liver is often subjected to oxidative stress that leads to lipid peroxidation and severe cellular damage. The conventional treatments of liver diseases such as cirrhosis, fatty liver and chronic hepatitis are frequently inadequate due to side effects caused by hepatotoxic chemical drugs. To overcome this problematic paradox, medicinal plants, owing to their natural richness in phenolic compounds, have been intensively exploited concerning their extracts and fraction composition in order to find bioactive compounds that could be isolated and applied in the treatment of liver ailments. The present review aimed to collect the main results of recent studies carried out in this field and systematize the information for a better understanding of the hepatoprotective capacity of medicinal plants in in vitro and in vivo systems. Generally, the assessed plant extracts revealed good hepatoprotective properties, justifying the fractionation and further isolation of phenolic compounds from different parts of the plant. Twenty-five phenolic compounds, including flavonoids, lignan compounds, phenolic acids and other phenolic compounds, have been isolated and identified, and proved to be effective in the prevention and/or treatment of chemically induced liver damage. In this perspective, the use of medicinal plant extracts, fractions and phenolic compounds seems to be a promising strategy to avoid side effects caused by hepatotoxic chemicals.

Ceratonia siliqua L. hydroethanolic extract obtained by ultrasonication: antioxidant activity, phenolic compounds profile and effects in yogurts functionalized with their free and microencapsulated forms

Bioactive extracts were obtained from powdered carob pulp through an ultrasound extraction process and then evaluated in terms of antioxidant activity. Ten minutes of ultrasonication at 375 Hz were the optimal conditions leading to an extract with the highest antioxidant effects. After its chemical characterization, which revealed the preponderance of gallotannins, the extract (free and microencapsulated) was incorporated in yogurts. The microspheres were prepared using an extract/sodium alginate ratio of 100/400 (mg mg(-1)) selected after testing different ratios. The yogurts with the free extract exhibited higher antioxidant activity than the samples added with the encapsulated extracts, showing the preserving role of alginate as a coating material. None of the forms significantly altered the yogurt's nutritional value. This study confirmed the efficiency of microencapsulation to stabilize functional ingredients in food matrices maintaining almost the structural integrity of polyphenols extracted from carob pulp and furthermore improving the antioxidant potency of the final product.

Wild mushrooms and their mycelia as sources of bioactive compounds: antioxidant, anti-inflammatory and cytotoxic properties

Mushrooms are an important source of natural compounds with acknowledged bioactivity. Pleurotus eryngii (DC.) Quél., in particular, is widely recognized for its organoleptic quality and favorable health effects, being commercially produced in great extent. On the other hand, Suillus bellinii (Inzenga) Watling is an ectomycorrhizal symbiont, whose main properties were only reported in a scarce number of publications. Some current trends point toward using the mycelia and the culture media as potential sources of bioactive compounds, in addition to the fruiting bodies. Accordingly, P. eryngii and S. bellinii were studied for their composition in phenolic acids and sterols, antioxidant capacity (scavenging DPPH radicals, reducing power, β-carotene bleaching inhibition and TBARS formation inhibition), anti-inflammatory effect (by down-regulating LPS-stimulated NO in RAW264.7 cells) and anti-proliferative activity (using MCF-7, NCI-H460, HeLa, HepG2 and PLP2 cell lines). Overall, S. bellinii mycelia showed higher contents of ergosterol and phenolic compounds (which were also detected in higher quantity in its fruiting body) and stronger antioxidant activity than P. eryngii. On the other hand, P. eryngii mycelia showed anti-inflammatory (absent in S. bellinii mycelia) and a cytotoxicity similar (sometimes superior) to its fruiting bodies, in opposition to S. bellinii, whose mycelia presented a decreased anti-proliferative activity. Furthermore, the assayed species showed differences in the growth rate and yielded biomass of their mycelia, which should also be considered in further applications.

HPLC-DAD-ESI/MS phenolic characterization and biological activity of Equisetum giganteum L.

Naturally-occurring phytochemicals have received a pivotal attention in the last years, due to the increasing evidences of biological activities. Equisetum giganteum L., commonly known as “giant horsetail”, is a native plant from Central and South America, being largely used in dietary supplements as diuretic, hemostatic, antiinflammatory and anti-rheumatic agents [1,2]. The aim of the present study was to evaluate the antioxidant (scavenging effects on 2,2-diphenyl-1-picrylhydrazyl radicals- RSA, reducing power- RP, β-carotene bleaching inhibition- CBI and lipid peroxidation inhibition- LPI), anti-inflammatory (inhibition of NO production in lipopolysaccharidestimulated RAW 264.7 macrophages) and cytotoxic (in a panel of four human tumor cell lines: MCF-7- breast adenocarcinoma, NCI-H460- non-small cell lung cancer, HeLa- cervical carcinoma and HepG2- hepatocellular carcinoma; and in non-tumor porcine liver primary cells- PLP2) properties of E. giganteum, providing a phytochemical characterization of its extract (ethanol/water, 80:20, v/v), by using highperformance liquid chromatography coupled to diode array detection and electrospray ionisation mass spectrometry (HPLC-DAD–ESI/MS). E. giganteum presented fourteen phenolic compounds, two phenolic acids and twelve flavonol glycoside derivatives, mainly kaempferol derivatives, accounting to 81% of the total phenolic content, being kaempferol-O-glucoside-O-rutinoside, the most abundant molecule (7.6 mg/g extract). The extract exhibited antioxidant (EC50 values = 123, 136, 202 and 57.4 μg/mL for RSA, RP, CBI and LPI, respectively), anti-inflammatory (EC50 value = 239 μg/mL) and cytotoxic (GI50 values = 250, 258, 268 and 239 μg/mL for MCF-7, NCI-H460, HeLa and HepG2, respectively) properties, which were positively correlated with its concentration in phenolic compounds. Furthermore, up to 400 μg/mL, it did not revealed toxicity in non-tumor liver cells. Thus, this study highlights the potential of E. giganteum extracts as rich sources of phenolic compounds that can be used in the food, pharmaceutical and cosmetic fields.

Comparative study of bioactive potential of Fueniculum vulgare Mill. and Matricaria recutita L. phenolic rich extracts to functionalize cottage chesse

Synthetic additives used in a wide variety of food products have been associated to some toxic effects. This conducted to an increasing interest of consumers for natural additives, including food preservers [1]. Many aromatic herbs have been used to prepare bioactive extracts with benefits to the consumer's health. Foeniculum vulgare Mill. (fennel) and Matricaria recutita L. (chamomile) are examples of popular herbs rich in phenolic compounds with documented antioxidant and antimicrobial properties [2,3]. The present work confirms the antioxidant (DPPH scavenging activity, reducing power and lipid peroxidation inhibition) and antimicrobial (against bacteria such as Bacillus cereus and Salmonella Typhimurium and fungi such as Aspergillus niger, A. versicolor and PenicilliumfimicuJosum) activities of fennel and chamomile extracts, obtained by decoction. The chemical characterization of the extracts, performed by HPLC-DAD-ESIIMS, revealed the presence of five flavonoids (mainly qercetin-3-0- glucoside) and twelve phenolic acids (mainly 5-0-caffeolyquinic acid) for fennel extract and the presence of nine flavonoids (mainly luteolin-0-glucuronide) and ten phenolic acids (mainly di-caffeoyl-2,7- anhydro-3-deoxy-2-octulopyranosonic acid) for chamomile extract. Due to their high antioxidant and antimicrobial activities, both extracts were then incorporated (at DPPH scavenging activity EC25 value: 0.35 mg/mL and 0.165 mg/mL for fennel and chamomile, respectively) in cottage cheeses (prepared by Queijos Casa Matias Lda) as natural additives with two objectives: to increase the shelf-life of the cottage cheeses and to provide bioactive properties to the final products. The results showed that the use of these natural extracts did not alter significantly the nutritional characteristics of the cottage cheese in comparison with control samples (cottage cheese without extracts), but improved its antioxidant potential (more evident in the samples with chamomile extract). After 14 days of storage, only the control samples showed signs of degradation. Overall, the present study highlights the preservation potential of fennel and chamomile extracts in cottage cheeses, improving also their bioactivity.