Development of a loop-mediated isothermal amplification assay for the rapid detection of Styphnolobium japonicum (L.) Schott as an adulterant of Ginkgo biloba (L.).

BACKGROUND: Species adulteration is a concern in herbal products, especially when plant substitutes of lower economic value replace valuable botanicals. Styphnolobium japonicum is well known as a potential adulterant of Ginkgo biloba, which is one of the most demanded medicinal plants due to its wide use in pharmaceuticals, food supplements, and traditional medicine. Despite bearing some resemblance to ginkgo's flavonol composition, S. japonicum lacks many of G. biloba's desired therapeutic properties. To prevent adulteration practices, it is crucial to implement rigorous quality control measures, including fast and simple diagnostic tools that can be used on-field. PURPOSE: This study aims to develop for the first time a species-specific loop-mediated isothermal amplification (LAMP) method for the fast identification of S. japonicum in ginkgo-containing products. METHODS: A set of four specific primers (SjF3, SjB3, SjFIP, and SjBIP) and loop primers (SjLF and SjLB) were designed for a LAMP based assay using the 5.8S partial sequence and the internal transcribed spacer 2 of nuclear ribosomal DNA of S. japonicum. RESULTS: The successful amplification of the LAMP assay was inspected through visual detection, with the highest intensity recorded at the optimal conditions set at 68 °C for 40 min. The primers showed high specificity and were able to accurately discriminate S. japonicum from G. biloba and 49 other species of medicinal plants. Furthermore, the proposed LAMP assay proved to be fast, selective, and highly sensitive, as demonstrated by the absolute and relative limits of detection, which were reached at 0.5 pg for S. japonicum DNA and 0.01 % S. japonicum in G. biloba, respectively. CONCLUSIONS: This novel approach allows easy identification and discrimination of S. japonicum as a potential adulterant of G. biloba, thus being a useful tool for quality control. Compared to chromatographic or PCR-based methods, the assay proved to be fast, sensitive and did not require expensive equipment, thus offering the possibly usage in field analysis.

Adulteration of Brain Health (Cognitive, Mood, and Sleep Enhancement) Food Supplements by the Addition of Pharmaceutical Drugs: A Comprehensive Review of Analytical Approaches and Trends.

In recent years, the consumption of dietary supplements has grown worldwide, particularly in developed regions. However, this growing market has also become a prime target for adulteration practices, with some manufacturers illegally adding pharmaceuticals into plant-based food supplements (PFS) to enhance their effects. While extensive research has focused on detecting adulterant drugs in PFS tailored for improving sexual performance, weight loss, and muscle building, less attention has been given to supplements intended for mood enhancement, sleep aid, and cognitive function (nootropics). Nonetheless, recent reports indicate an increasing level of adulteration within this group of PFS. Therefore, this review aims at providing a comprehensive overview on the adulteration of PFS tailored for brain health, with a focus on the analytical techniques utilized for detection while also presenting data on consumption patterns and the prevalence of reported adulterants. Considering that the detection of such fraudulent practices primarily relies on chromatographic techniques coupled with mass spectrometry (MS), the developments in this field comprising either targeted or untargeted analysis of pharmaceutical adulterants are discussed.

Green Extraction Strategy Using Bio-Based Aqueous Biphasic Systems for Polyphenol Valorization from Grape By-Product.

Polyphenols are natural compounds with enhanced antioxidant properties. They are present in relatively high concentrations in fruit/vegetable by-products. Therefore, there is a need for the development of efficient and cost-effective methods for the separation and purification of these valuable compounds. Traditional extraction with organic solvents needs to be switched to novel methods that are more efficient, with reduced extraction times and low consumption of organic solvents. Aiming at developing sustainable processes for the separation and purification of phenolic compounds, we used three model compounds, namely resveratrol, quercetin, and gallic acid, to investigate ionic liquid-based aqueous biphasic systems (IL-ABSs) formed by cholinium-based IL in combination with polypropylene glycol with a molecular mass of 400 g/mol (PPG400). The ABS composition in the two-phase region was selected according to a previously determined phase diagram. Extraction studies indicated the preferential partition of resveratrol and quercetin toward the hydrophobic PPG-rich phase that is mainly dominated by its hydrophobic nature and the strong salting-out effect of ILs. On the other hand, due to its considerably hydrophilic nature, gallic acid preferentially migrates toward the IL phase. The achieved results from grape stem extract demonstrated high extraction efficiencies of cholinium dihydrogen phosphate (~99% for resveratrol for the PPG phase and 78% for gallic acid for the IL phase), with considerable selectivity, demonstrating promising outcomes for potential applications.

Disclosing the potential of Cupressus leylandii A.B. Jacks & Dallim, Eucalyptus globulus Labill., Aloysia citrodora Paláu, and Melissa officinalis L. hydrosols as eco-friendly antimicrobial agents.

Antimicrobial resistance is a major global health concern, threatening the effective prevention and treatment of infections caused by microorganisms. These factors boosted the study of safe and green alternatives, with hydrosols, the by-products of essential oils extraction, emerging as promising natural antimicrobial agents. In this context, four hydrosols obtained from Cupressus leylandii A.B. Jacks & Dallim, Eucalyptus globulus Labill., Aloysia citrodora Paláu and Melissa officinalis L. were studied. Their chemical composition comprises neral, geranial, 1,8-cineole, terpinen-4-ol, and oplopanonyl acetate, compounds with recognised antimicrobial activity. Concerning antimicrobial activity, significant differences were found using different hydrosol concentrations (10-20% v/v) in comparison to a control (without hydrosol), showing the potential of the tested hydrosols to inhibit the microbial growth of Escherichia coli, Staphylococcus aureus, and Candida albicans. A. citrodora hydrosol was the most effective one, inhibiting 90% of E. coli growth and 80% of C. albicans growth, for both hydrosol concentrations (p < 0.0001). With hydrosol concentration increase, it was possible to observe an improved antimicrobial activity with significant reductions (p < 0.0001). The findings of this work indicate the viability of reusing and valuing the hydrosols, encouraging the development of green applications for different fields (e.g., food, agriculture, pharmaceuticals, and cosmetics).

Mass spectrometry-based approaches to assess the botanical authenticity of dietary supplements.

Dietary supplements are legally considered foods despite frequently including medicinal plants as ingredients. Currently, the consumption of herbal dietary supplements, also known as plant food supplements (PFS), is increasing worldwide and some raw botanicals, highly demanded due to their popularity, extensive use, and/or well-established pharmacological effects, have been attaining high prices in the international markets. Therefore, botanical adulteration for profit increase can occur along the whole PFS industry chain, from raw botanicals to plant extracts, until final PFS. Besides the substitution of high-value species, unintentional mislabeling can happen in morphologically similar species. Both cases represent a health risk for consumers, prompting the development of numerous works to access botanical adulterations in PFS. Among different approaches proposed for this purpose, mass spectrometry (MS)-based techniques have often been reported as the most promising, particularly when hyphenated with chromatographic techniques. Thus, this review aims at describing an overview of the developments in this field, focusing on the applications of MS-based techniques to targeted and untargeted analysis to detect botanical adulterations in plant materials, extracts, and PFS.

Screening of Chemical Composition, Antimicrobial and Antioxidant Activities in Pomegranate, Quince, and Persimmon Leaf, Peel, and Seed: Valorization of Autumn Fruits By-Products for a One Health Perspective.

Antimicrobial resistance is increasing globally and is now one of the major public health problems. Therefore, there is a need to search for new antimicrobial agents. The food industry generates large amounts of by-products that are rich in bioactive compounds, such as phenolic compounds, which are known to have several health benefits, including antioxidant and antimicrobial properties. Thus, we aimed to characterize the phenolic compounds present in pomegranate, quince, and persimmon by-products, as well as their antioxidant and antimicrobial activities. Phenolic compounds were extracted from pomegranate, quince, and persimmon leaves, seeds, and peels using a mixture of ethanol/water (80/20). The polyphenol profile of the extracts was determined by high-performance liquid chromatography. The antioxidant activity of the extracts was determined by the 2,2-diphenyl-1-picrylhydrazyl (DPPH), ferric reducing antioxidant power (FRAP), and cupric reducing antioxidant capacity (CUPRAC) methods. Antimicrobial susceptibility was evaluated using the Kirby-Bauer disk diffusion method. In general, leaves showed higher concentrations of phenolics than the peel and seeds of fruits. In total, 23 phenolic compounds were identified and quantified, with sanguiin and apigenin-3-O-galactoside being present in the highest concentrations. Leaf extracts of pomegranate showed higher antioxidant activities than the other components in all methods used. In general, all extracts had a greater antimicrobial activity against Gram-positive bacteria. Persimmon leaf and seed extracts inhibited a greater number of bacteria, both Gram-positive and -negative. The lowest minimum inhibitory concentration (MIC) detected among Gram-positive and -negative bacteria was 10 mg/mL for pomegranate peel and leaf extracts against Staphylococcus aureus and S. pseudintermedius and for pomegranate leaf extract against Escherichia coli. Our results reinforce the need to value food industry by-products that could be used as food preservatives and antibiotic adjuvants against multiresistant bacteria.

Yield, Chemical Composition and Bioactivity of Essential Oils from Common Juniper (Juniperus communis L.) from Different Spanish Origins.

Essential oils (EOs) obtained from Juniperus communis L. are frequently used in the production of bioproducts. However, there are no studies regarding industrial crops' production, allowing for better control of the quality and production of juniper EOs. To select the plant material for developing future crops of this species in northern Spain, four locations where this shrub species grows in the wild were selected and samples of both genera were collected. The EOs were obtained by steam distillation, and their chemical composition and bioactivity were evaluated. The results showed that the yield of EOs from male and female samples were within the usual reported ranges, varying between 0.24 and 0.58% (dry basis, d.b.). However, limonene content in three locations varied between 15 and 25%, which is between 100% and 200% higher than the values usually reported for other European countries. The antibacterial activity was determined by broth microdilution and showed that gram-positive bacteria were more susceptible to the tested EOs since, in general, lower MIC values were obtained compared to gram-negatives. The EOs from location 1 (L1F) and 2 (L2M) inhibited the growth of six out of the eight clinical strains tested. Samples from location 1 were particularly effective, exhibiting MBC against two gram-negative (E. coli and P. mirabilis) and one gram-positive bacteria (E. faecalis). Moreover, the majority of the EOs tested showed anti-inflammatory activity. Cytotoxic effect has been demonstrated in tumor cell lines, with the best results observed against gastric carcinoma (AGS) cells (GI50 between 7 to 77 µg/mL). Although generally presenting higher GI50, most samples also inhibited the growth of non-tumoral cells, particularly hepatocytes (PLP2 cells). Therefore, its use for their anti-proliferative activity should consider specific conditions to avoid damaging normal cells. Finally, the results and conclusions obtained led to the selection of the female shrubs from location 1 (L1F) as the plant material to be propagated in order to produce plants for a future juniper crop.

Volatile Compounds and Biological Activity of the Essential Oil of Aloysia citrodora Paláu: Comparison of Hydrodistillation and Microwave-Assisted Hydrodistillation.

Aromatic plants are a remarkable source of natural products. Aloysia citrodora Paláu (Verbenaceae), commonly known as lemon verbena, is a relevant source of essential oils with potential applications due to its lemony scent and bioactive properties. Studies carried out on this species have focused on the volatile composition of the essential oil obtained by Clevenger hydrodistillation (CHD), with little information available on alternative extraction methodologies or the biological properties of the oil. Therefore, this work aimed to compare the volatile composition, antioxidant activity, cytotoxicity, anti-inflammatory and antibacterial activities of the essential oil extracted by conventional hydrodistillation by Clevenger (CHD) and Microwave-Assisted Hydrodistillation (MAHD). Significant differences (p < 0.05) were observed for some compounds, including the two major ones, geranial (18.7-21.1%) and neral (15.3-16.2%). Better antioxidant activity was exhibited by the MAHD essential oil in DPPH radical scavenging and reducing power assays, while no differences were observed in the cellular antioxidant assay. The MADH essential oil also presented higher inhibition against four tumoral cell lines and exhibited lower cytotoxicity in non-tumoral cells as compared with Clevenger-extracted essential oil. In contrast, the latter showed higher anti-inflammatory activity. Both essential oils were able to inhibit the growth of eleven out of the fifteen bacterial strains tested.

Assessment of the Use of Common Juniper (Juniperus communis L.) Foliage following the Cascade Principle.

Juniperus communis L. is a species commonly grown in regions of the Northern Hemisphere, and is a good candidate to be cultivated in marginal lands. Plants coming from a pruning performed in a natural population located in Spain were used to assess the yield and quality of different products obtained following the cascade principle. A total of 1050 kg of foliage biomass were crushed, steam-distilled, and separated into fractions to produce biochar and absorbents for the pet industry using pilot plants. The obtained products were analysed. The essential oil, with a yield of 0.45% dry basis and a qualitative chemical composition similar to that described for the berries in international standards or monographs, showed antioxidant activity with promising CAA results (inhibition of 89% of the cell's oxidation). However, regarding antibacterial and antifungal activities, it only inhibited the growth of microorganisms at the maximum concentration tested, 2.5%. Concerning the hydrolate, it did not show bioactivity. Regarding the biochar, whose yield was 28.79% dry basis, interesting results were obtained for its characterisation as a possible soil improver for agronomic purposes (PFC 3(A)). Finally, promising results were obtained regarding the use of common juniper as absorbent, taking into account the physical characterisation and odour control capacity.

Essential Oils from Côa Valley Lamiaceae Species: Cytotoxicity and Antiproliferative Effect on Glioblastoma Cells.

Lavandula pedunculata (Mill.) Cav., Mentha cervina L. and Thymus mastichina (L.) L. subsp. mastichina are representative species of the Côa Valley's flora, a Portuguese UNESCO World Heritage Site. L. pedunculata and T. mastichina are traditionally used to preserve olives and to aromatize bonfires on Saint John's Eve, while M. cervina is mainly used as a spice for river fish dishes. Despite their traditional uses, these aromatic plants are still undervalued, and literature regarding their bioactivity, especially anticancer, is scarce. In this work, the morphology of secretory structures was assessed by scanning electron microscopy (SEM), and the composition of essential oils (EOs) was characterized by gas chromatography-mass spectrometry (GC-MS). The study proceeded with cytotoxic evaluation of EOs in tumor and non-tumor cells with the cell death mechanism explored in glioblastoma (GB) cells. L. pedunculata EO presented the most pronounced cytotoxic/antiproliferative activity against tumor cells, with moderate cytotoxicity against non-tumor cells. Whereas, M. cervina EO exhibited a slightly lower cytotoxic effect against tumor cells and did not affect the viability of non-tumor cells. Meanwhile, T. mastichina EO did not induce a strong cytotoxic effect against GB cells. L. pedunculata and M. cervina EOs lead to cell death by inducing apoptosis in a dose-dependent manner. The present study suggests that L. pedunculata and M. cervina EOs have a strong cytotoxic and antiproliferative potential to be further studied as efficient antitumor agents.

Chemical Composition and Biological Activity of Commelina erecta: An Edible Wild Plant Consumed in Brazil.

In recent years, the interest in products of natural origin has boosted the exploitation and use of plants as food and sources of bioactive compounds, especially wild plants widely used in different cultures for several purposes. Commelina erecta is a wild edible plant (WEP) traditionally used as food and medicine, about which few studies exist. Thus, this study aimed at enhancing the knowledge about its nutritional, chemical and bioactive profile, considering different plant parts and development stages, in order to increase its inclusion in the diet of South American communities. The nutritional profile was found to be similar to other WEP frequently consumed in Brazil. Thirteen phenolic compounds (HPLC-DAD-ESI/MS) were tentatively identified, with apigenin, luteolin and quercetin derivatives being the most abundant. Fructose and oxalic acid were the major sugar and organic acid, respectively, in the aerial parts of C. erecta, and four isoforms of tocopherols were also identified. Regarding the plant's antioxidant activity, the EC50 values varied between 18.4 and 1060 µg/mL in the inhibition of lipid peroxidation assay (TBARS) and between 53 and 115 µg/mL in the oxidative haemolysis inhibition (OxHLIA) assay. The hydroethanolic extract obtained from stems at the flowering stage also presented anti-inflammatory activity. In general, all the extracts evidenced promising antimicrobial activity. Altogether, these results reinforce the traditional use of this plant species as food and medicine to support the diet of needier populations and also promote food sovereignty and sustainability.

Authentication of incense (Pittosporum undulatum Vent.) honey from the Azores (Mel dos Açores) by a novel real-time PCR approach.

'Mel dos Açores' is a unique nectar honey produced from the exceptional and diverse flora of the Azores archipelago, categorised as incense honey ('mel de incenso') or multifloral honey ('mel multiflora'). Incense honey should contain over 30 % of pollen grains of Pittosporum undulatum Vent. In this work, a real-time PCR method targeting the ITS region was proposed for the first time to detect P. undulatum in the honey from the Azores. The approach exhibited high analytical performance, achieving a quantification limit of 0.01 pg of incense DNA. The method was successfully applied to 22 honey samples, from which incense was detected in all 9 monofloral incense honeys and in 5 out of 10 multifloral samples from the Azores. Generally, the quantitative results for incense DNA were in good agreement with the melissopalynological data. Therefore, a simple, cost-effective and reliable tool was herein proposed to authenticate and valorise the Azores honey.

A novel real-time PCR coupled with high resolution melting analysis as a simple and fast tool for the entomological authentication of honey by targeting Apis mellifera mitochondrial DNA.

Honey is one of the foods easily adulterated worldwide. Recently, the analysis of honeybee DNA has been proposed as a useful tool to authenticate the entomological origin of honey. However, the methods proposed so far require more than one polymerase chain reaction (PCR) and the use of agarose gels, making the authentication process laborious and lengthy. In this work, a novel real-time PCR coupled with high-resolution melting (HRM) analysis of a 150 bp fragment of the cytochrome c oxidase I (COI) gene is proposed as a fast and simple tool to assess honey's entomological origin by discriminating the mitochondrial DNA lineages of European honey bees (A, M and C lineages). In addition, the new tool allowed the differentiation of honeys produced by different mitotypes of C-lineage ancestry. The method showed high analytical performance and was able to successfully identify the entomological origin of honeys of known origin obtained from research apiaries/beekeepers. Therefore, it was applied to 44 commercial honeys from different countries. It confirmed the entomological authenticity of French PDO honeys that should be produced by the Corse ecotype A. m. mellifera. For the remaining honeys, the results were also in good agreement with the declared geographical origin. However, three honeys from Slovenia did not cluster with C2 mitotype A. m. carnica as expected, suggesting the mixture of honeys produced by honeybees of different mitotypes.

Comparative study on the phenolic composition and in vitro bioactivity of medicinal and aromatic plants from the Lamiaceae family.

Medicinal and aromatic plants (MAP) have been described as a source of phenolic compounds with potential as antioxidant, antiproliferative and antimicrobial agents. MAP from the Lamiaceae family (Origanum vulgare L., Thymus vulgaris L., Ocimum basilicum L., Salvia officinalis L., Melissa officinalis L., and MatricariachamomillaL.) were selected to perform a phytochemical and biological screening for their further exploitation as natural bioactive ingredients. The total content of phenolic compounds varied from 184.02 mg/g extract in M. officinalis to 17.97 mg/g extract in M. chamomilla. Caffeic and rosmarinic acids were the main phenolic acids found in the respective hydroalcoholic extracts. The extracts showed a promising antioxidant activity in vitro, being related the phenolic compositions of the extracts, furthermore, all extracts being able to combat lipid peroxidation in TBARS assays with an IC50 under 26 µg/mL, moreover all the plant extract has prevented the oxidative haemolysis in OxHLIA assays at concentrations below 67 µg/mL in a Δt 60 min and under 118 µg/mL for a Δt 120 min. Regarding to the bactericidal and fungicidal action the plant extracts were able to inhibit growth against bacteria associated with food hazards, such as Salmonella typhimurium (MIC < 1) and Listeria monocytogenes (MIC < 1), regarding to fungicidal activity it can be highlighted the MIC values under to 0.25 for Aspergillus versicolor and Trichoderma viride. Overall, the selected Lamiaceae plants stood out as a source of active phytochemicals that can be used by different industries, such as food and cosmetics.

Analysis of a Vegetable Oil Performance in a Milling Process by MQL Lubrication.

In this work, we carried out a comparison between the dry machining of an aluminum block with conventional cutting oil and a block with vegetable oil. The two oils had different flow rates. Using the Taguchi method, it was possible to determine the matrices for optimizing the best parameters for each group of tests. Then, we studied the utility of using vegetable oil as a cutting lubricant. We found that the vegetable oil studied in this work had good properties in terms of reducing cutting temperatures but was less effective than conventional cutting oil in reducing the surface roughness of the machined part. Tribological tests were carried out to understand the influence of the selected lubricants in reducing friction and wear. After the sliding experiments, which were performed without lubrication in the presence of the same lubricants that were used in the machining tests and in the presence of distilled water, we concluded that vegetable oil has satisfactory lubricating properties that are similar to those of the conventional cutting fluid, indicating a potential for consideration as an effective alternative to the conventional cutting fluid, with economic, environmental, and health advantages.

Authentication of Argan (Argania spinosa L.) Oil Using Novel DNA-Based Approaches: Detection of Olive and Soybean Oils as Potential Adulterants.

Argan oil is a traditional product obtained from the fruits of the argan tree (Argania spinosa L.), which is endemic only to Morocco. It is commercialized worldwide as cosmetic and food-grade argan oil, attaining very high prices in the international market. Therefore, argan oil is very prone to adulteration with cheaper vegetable oils. The present work aims at developing novel real-time PCR approaches to detect olive and soybean oils as potential adulterants, as well as ascertain the presence of argan oil. The ITS region, matK and lectin genes were the targeted markers, allowing to detect argan, olive and soybean DNA down to 0.01 pg, 0.1 pg and 3.2 pg, respectively, with real-time PCR. Moreover, to propose practical quantitative methods, two calibrant models were developed using the normalized ΔCq method to estimate potential adulterations of argan oil with olive or soybean oils. The results allowed for the detection and quantification of olive and soybean oils within 50-1% and 25-1%, respectively, both in argan oil. Both approaches provided acceptable performance parameters and accurate determinations, as proven by their applicability to blind mixtures. Herein, new qualitative and quantitative PCR assays are proposed for the first time as reliable and high-throughput tools to authenticate and valorize argan oil.

Animal Species Authentication in Dairy Products.

Milk is one of the most important nutritious foods, widely consumed worldwide, either in its natural form or via dairy products. Currently, several economic, health and ethical issues emphasize the need for a more frequent and rigorous quality control of dairy products and the importance of detecting adulterations in these products. For this reason, several conventional and advanced techniques have been proposed, aiming at detecting and quantifying eventual adulterations, preferentially in a rapid, cost-effective, easy to implement, sensitive and specific way. They have relied mostly on electrophoretic, chromatographic and immunoenzymatic techniques. More recently, mass spectrometry, spectroscopic methods (near infrared (NIR), mid infrared (MIR), nuclear magnetic resonance (NMR) and front face fluorescence coupled to chemometrics), DNA analysis (real-time PCR, high-resolution melting analysis, next generation sequencing and droplet digital PCR) and biosensors have been advanced as innovative tools for dairy product authentication. Milk substitution from high-valued species with lower-cost bovine milk is one of the most frequent adulteration practices. Therefore, this review intends to describe the most relevant developments regarding the current and advanced analytical methodologies applied to species authentication of milk and dairy products.

Phytic Acid against Clostridium perfringens Type A: A Food Matrix Study.

This study evaluated the inhibitory effect of phytic acid (PA) on the spore germination and vegetative cells growth of Clostridium perfringens type A, as well as its effect in combination with maltodextrin (MD) in cooked sausages. The addition of 1% PA showed a satisfactory inhibition of spores' germination and vegetative cells growth of C. perfringens in BHI media. The inhibitory effect of 1% PA on vegetative cells was similar to the additive sodium sorbate (SS) at 10%. Subsequently, a mixture of PA-MD (1:1; w/w) was evaluated for the inhibition of C. perfringens spores in cooked sausages. The PA-MD 1.5% and 2.5% had a similar performance to SS 10% and a similar or higher performance than 0.015% NO2 (p < 0.05). In an unprecedented way, the present study demonstrated that PA inhibited spore germination and vegetative cells growth of C. perfringens, highlighting its potential use as an alternative and natural preservative for the meat industry.

Botanical authentication of globe artichoke-containing foods: Differentiation of Cynara scolymus by a novel HRM approach.

Cynara scolymus L., known as globe artichoke, is a medicinal plant widely used in plant food supplements (PFS) and herbal infusions due to its beneficial health properties. The high demand for artichoke-containing products can lead to adulteration practices. In this work, a real-time polymerase chain reaction (PCR) system coupled to high-resolution melting (HRM) analysis was proposed to differentiate C. scolymus from other Cynara species. Hence, a Cynara-specific real-time PCR assay was successfully developed with high analytical performance, achieving a sensitivity of 0.4 pg of globe artichoke DNA. HRM analysis enabled the discrimination of C. scolymus, with a high level of confidence (>98%), corroborating sequencing data. Application results to artichoke-containing PFS and mixed herbal infusions allowed confirming the presence of C. scolymus in 38% of the samples, suggesting the substitution/mislabelling of globe artichoke in 2 samples and the need for further efforts to increase DNA amplifiability of PFS.

Chemical and Bioactive Characterization of the Essential Oils Obtained from Three Mediterranean Plants.

Cupressus sempervirens L., Juniperus communis L. and Cistus ladanifer L. are Mediterranean arboreal and shrub species that possess essential oils (EO) in their leaves and branches. This study aimed at characterizing the EOs obtained by steam distillation from the three species collected in different locations from Spain (Almazán, Andévalo, Barriomartín, Cerezal, Ermitas and Huéscar). For this purpose, volatiles composition was determined by GC-MS, and different bioactivities were evaluated. The highest content in terpenes was observed in C. sempervirens (Huéscar origin) followed by J. communis (Almazán origin), corresponding to 92% and 91.9% of total compounds, respectively. With exception of C. ladanifer from Cerezal that presented viridiflorol as the most abundant compound, all the three species presented in common the α-pinene as the major compound. The EOs from C. ladanifer showed high antibacterial potential, presenting MIC values from 0.3 to 1.25 mg/mL. Concerning other bioactivities, C. ladanifer EO revealed an oxidation inhibition of 83%, while J. communis showed cytotoxicity in the MCF-7 cell line, and C. sempervirens and C. ladanifer EOs exhibited the highest potential on NCI-H460 cell lines. Nevertheless, some EOs revealed toxicity against non-tumoral cells but generally presented a GI50 value higher than that of the tumor cell lines.