Analysis of Bioactive Aroma Compounds in Essential Oils from Algerian Plants: Implications for Potential Antioxidant Applications.

In samples of Artemisia campestris (AC), Artemisia herba-alba (AHA) and Salvia jordanii (SJ) essential oils, up to 200 distinct volatile compounds were identified. Using headspace solid-phase microextraction combined with gas chromatography-olfactometry-mass spectrometry (HS-SPME-GC-O-MS), different panelists detected 52 of these compounds. This study offers the most detailed analysis of bioactive compound profiles conducted so far. The most abundant compounds identified were curcumene, making up 12.96% of AC, and camphor, constituting 21.67% of AHA and 19.15% of SJ. The compounds with the highest odor activity value (OAV) were (E,Z)-2,4-nonadienal (geranium, pungent), 3-nonenal (cucumber) and 2-undecenal (sweet) in AC, AHA and SJ, respectively. AHA essential oil showed significant antioxidant activity (IC50 = 41.73 ± 4.14 mg/g) and hydroxyl radical generation (hydroxylation percentage = 29.62 ± 3.14), as assessed by the diphenylpicrylhydrazyl (DPPH) method. In terms of oxygen radical absorbance capacity (ORAC), the strongest antioxidant activity was obtained for SJ essential oil (antioxidant activity of the essential oils, AOX = 337.49 ± 9.87).

Phenolic compounds profile of macerates of different edible parts of carob tree (Ceratonia siliqua L.) using UPLC-ESI-Q-TOF-MSE: Phytochemical screening and biological activities.

Locust bean pulp and gum extracts were prepared, and phytochemical tests based on color reactions and chromatographic analyzes were performed. A profile of seventy-six phenolic compounds was obtained by the ultra-high performance liquid chromatography with electrospray ionization and quadrupole time-of-flight mass spectrometry. The main groups of phenolic compounds identified in the both extracts of Ceratonia siliqua L., were flavonoids, tannins and phenolic acids. Moreover, carob pulp and gum extracts were tested for their antimicrobial activity using disk diffusion tests which showed sensitivity of the different strains to the analyzed extracts at a concentration of 100 mg/mL. Additionally, the antioxidant activity of Ceratonia siliqua L. extracts was assessed by the 2,2-diphenyl-1-picrylhydrazyl acid test, which confirmed stronger antioxidant properties in the case of the pulp extract. To sum up, carob pulp and gum extracts present promising alternatives to synthetic additives within the medicinal industry, serving as potential antioxidant agents and preservatives that combat bacterial contamination, thereby offering a more natural approach to enhancing product safety and efficacy.

Evaluation the Potential of Onion/Laponite Composites Films for Sustainable Food Packaging with Enhanced UV Protection and Antioxidant Capacity.

Agro-industrial residues have attracted attention for their applications in the field of biodegradable packaging. Recently, our research group has developed onion-based films with promising properties for this type of application due to their non-toxicity, biocompatibility and biodegradability. Therefore, in this study, we investigated the effect of Laponite clay concentration on the physicochemical and antioxidant properties of the onion-based films, which were prepared by a casting method. The XRD and FTIR data confirm the presence of the mineral clay in the onion-based films. These findings are consistent with those obtained from FE-SEM analysis, which revealed the presence of typical Laponite grains. In terms of wettability, the results show that the clay decreases the hydrophilic character of the material but slightly increases the water vapor permeation. Optical characterization revealed that the materials exhibited zero transmittance in the UV region and increased opacity in the visible region for composites containing 5% and 10% Laponite. Furthermore, the antioxidant test demonstrated higher antioxidant potential in the composites compared to the pure films. Consequently, these results suggest that the formation of Laponite and onion composites could be an essential strategy for developing natural polymers in the field of food contact packaging.

Ultra-high performance liquid chromatography coupled to ion mobility quadrupole time-of-flight mass spectrometry for the identification of non-volatile compounds migrating from 'natural' dishes.

Although most new biomaterials for food industry applications are labelled '100% natural fabrication' and 'chemical-free', certain compounds may migrate from those materials to the food, compromising the organoleptic characteristics and safety of the product. In this work, the degree of compound migration from dishes made with four different biomaterials: bamboo, palm leaf, wood and wheat pulp was investigated. Migration tests were carried out using three food simulants, 10% ethanol (simulant A), 3% acetic acid (simulant B), and 95% ethanol (simulant D2). Unequivocal identification of non-intentionally added substances (NIAS) is challenging even when using high-resolution mass spectrometry techniques however, a total of 25 different non-volatile compounds from the migration tests were identified and quantified using Ultra-high performance liquid chromatography coupled to ion mobility quadrupole time-of-flight mass spectrometry (UPLC-IMS-MS). In the bamboo samples three oligomers, cyclic diethylene glycol adipate, 3,6,9,16,19,22-hexaoxabicyclo[22.3.1]-octacosa-1(28),24,26-triene-2,10,15,23-tetrone and 1,4,7,14,17,20-hexaoxacyclohexacosane-8,13,21,26-tetrone exceeded the specified limits of migration.

Tumor Mutational Burden as a Predictor of Survival with Durvalumab and/or Tremelimumab Treatment in Recurrent or Metastatic Head and Neck Squamous Cell Carcinoma.

PURPOSE: Biomarkers that predict response to immune checkpoint inhibitors (ICI) in recurrent or metastatic head and neck squamous cell carcinoma (R/M HNSCC) are needed. This retrospective study assessed tumor mutational burden (TMB) and outcomes in the phase II HAWK and CONDOR and phase III EAGLE studies of durvalumab with or without tremelimumab in platinum-resistant R/M HNSCC. PATIENTS AND METHODS: Tumor samples from HAWK/CONDOR (N = 153) and blood samples from EAGLE (N = 247) were analyzed for TMB. Associations with survival were evaluated for tissue TMB (tTMB) at cutoffs from 10 to 20 mutations/megabase (mut/Mb) and for blood plasma TMB (bTMB) at cutoffs from 8 to 24 mut/Mb. RESULTS: In HAWK/CONDOR, overall survival (OS) with durvalumab with or without tremelimumab was longer for high versus low tTMB: statistically significant differences were observed with durvalumab plus tremelimumab at tTMB ≥ 10 mut/Mb [HR, 0.52 (95% confidence interval, CI, 0.28-0.98)] and tTMB ≥ 12 mut/Mb [HR, 0.46 (95% CI, 0.24-0.86)]. In EAGLE, a significant OS benefit versus chemotherapy was observed with durvalumab and durvalumab plus tremelimumab at bTMB≥16 mut/Mb [HR, 0.39 (95% CI, 0.20-0.76) and 0.38 (95% CI, 0.19-0.78), respectively] but not bTMB < 16 mut/Mb [HR, 0.92 (0.61-1.37) and 0.92 (95% CI, 0.62-1.36), respectively]. A significant progression-free survival benefit was also observed in the ICI arms versus chemotherapy at bTMB ≥ 16 mut/Mb. CONCLUSIONS: Findings support TMB as a biomarker for predicting survival in patients with platinum-resistant R/M HNSCC treated with ICIs. The analysis of EAGLE demonstrated that bTMB was predictive of survival with ICI treatment versus chemotherapy in a large, randomized controlled study population.

Promoter-pervasive transcription causes RNA polymerase II pausing to boost DOG1 expression in response to salt.

Eukaryotic genomes are pervasively transcribed by RNA polymerase II. Yet, the molecular and biological implications of such a phenomenon are still largely puzzling. Here, we describe noncoding RNA transcription upstream of the Arabidopsis thaliana DOG1 gene, which governs salt stress responses and is a key regulator of seed dormancy. We find that expression of the DOG1 gene is induced by salt stress, thereby causing a delay in seed germination. We uncover extensive transcriptional activity on the promoter of the DOG1 gene, which produces a variety of lncRNAs. These lncRNAs, named PUPPIES, are co-directionally transcribed and extend into the DOG1 coding region. We show that PUPPIES RNAs respond to salt stress and boost DOG1 expression, resulting in delayed germination. This positive role of pervasive PUPPIES transcription on DOG1 gene expression is associated with augmented pausing of RNA polymerase II, slower transcription and higher transcriptional burst size. These findings highlight the positive role of upstream co-directional transcription in controlling transcriptional dynamics of downstream genes.

Single seeds exhibit transcriptional heterogeneity during secondary dormancy induction.

Seeds are highly resilient to the external environment, which allows plants to persist in unpredictable and unfavorable conditions. Some plant species have adopted a bet-hedging strategy to germinate a variable fraction of seeds in any given condition, and this could be explained by population-based threshold models. Here, in the model plant Arabidopsis (Arabidopsis thaliana), we induced secondary dormancy (SD) to address the transcriptional heterogeneity among seeds that leads to binary germination/nongermination outcomes. We developed a single-seed RNA-seq strategy that allowed us to observe a reduction in seed transcriptional heterogeneity as seeds enter stress conditions, followed by an increase during recovery. We identified groups of genes whose expression showed a specific pattern through a time course and used these groups to position the individual seeds along the transcriptional gradient of germination competence. In agreement, transcriptomes of dormancy-deficient seeds (mutant of DELAY OF GERMINATION 1) showed a shift toward higher values of the germination competence index. Interestingly, a significant fraction of genes with variable expression encoded translation-related factors. In summary, interrogating hundreds of single-seed transcriptomes during SD-inducing treatment revealed variability among the transcriptomes that could result from the distribution of population-based sensitivity thresholds. Our results also showed that single-seed RNA-seq is the method of choice for analyzing seed bet-hedging-related phenomena.

Polylactide-Based Films with the Addition of Poly(ethylene glycol) and Extract of Propolis-Physico-Chemical and Storage Properties.

Polymeric films based on polylactide (PLA) with the addition of poly(ethylene glycol) (PEG) and a chloroformic extract of propolis were obtained. In the case of the studied films, polylactide (PLA) played the role of polymeric matrix and poly(ethylene glycol) was used as a plasticizer, while the extract of propolis was incorporated as a compound that could significantly affect the properties of the obtained materials, especially the water vapour permeation rate and the stability of the food products. Moreover, changes in structure, morphology, mechanical and storage properties as well as differences in colour, thickness and transparency after introducing propolis into the PLA-PEG system were determined. Based on the obtained results, it was established that the addition of the chloroformic extract of propolis significantly influences the most important properties taken into account during food packaging. It was also noticed that films with incorporated propolis were characterised by a significant improvement in the water vapour barrier property. Moreover, the obtained results prove that packaging containing a chloroformic propolis extract allow for the maintenance of the quality of the fruit stored for an extended period of time. To summarise, the application of a chloroformic propolis extract enables the formation of packaging materials that extend the shelf life of stored food products.

Polylactide-Based Films Incorporated with Berberine-Physicochemical and Antibacterial Properties.

A series of new polymeric materials consisting of polylactide (PLA), polyethylene glycol (PEG) and berberine chloride (B) was evaluated. PEG was incorporated into the polymer matrix with the aim of obtaining a plasticizing effect, while berberine was added in order to obtain antibacterial properties in formed packaging materials. Materials were formed using the solvent-casting procedure. Fourier transform infrared spectroscopy and scanning electron microscopy were used so as to establish the structural changes resulting from the introduction of berberine. Thermogravimetry and differential scanning calorimetry were applied to study the thermal properties. Further, mechanical properties and differences in colour and transparency between the control sample and films containing berberine were also studied. The recorded data indicates that berberine formed a network on the surface of the PLA-based materials. Introduction of an active compound significantly improved thermal stability and greatly affected the Young's modulus values of the studied polymeric films. Moreover, it should be stressed that the addition of the studied active compound leads to an improvement of the antibacterial properties, resulting in a significant decrease in growth of E. coli and the S. aureus bacteria cultures.

Electrospun Nanosystems Based on PHBV and ZnO for Ecological Food Packaging.

The electrospun nanosystems containing poly(3-hydroxybutyrate-co-3-hydroxyvalerate) (PHBV) and 1 wt% Fe doped ZnO nanoparticles (NPs) (with the content of dopant in the range of 0-1 wt% Fe) deposited onto polylactic acid (PLA) film were prepared for food packaging application. They were investigated by scanning electron microscopy (SEM), energy dispersive X-ray (EDX), Fourier transform infrared spectroscopy (FT-IR), X-ray diffraction (XRD), antimicrobial analysis, and X-ray photoelectron spectrometry (XPS) techniques. Migration studies conducted in acetic acid 3% (wt/wt) and ethanol 10% (v/v) food simulants as well as by the use of treated ashes with 3% HNO3 solution reveal that the migration of Zn and Fe falls into the specific limits imposed by the legislation in force. Results indicated that the PLA/PHBV/ZnO:Fex electrospun nanosystems exhibit excellent antibacterial activity against the Pseudomonas aeruginosa (ATCC-27853) due to the generation of a larger amount of perhydroxyl (˙OOH) radicals as assessed using electron paramagnetic resonance (EPR) spectroscopy coupled with a spin trapping method.

Importance of profile of volatile and off-odors compounds from different recycled polypropylene used for food applications.

Nowadays, polypropylene is one of the most common polymers used in the food packaging industry due to its good functionality and relatively low cost. Nevertheless, usage of plastic disposable packaging can be a generator of plastic pollution having negative environmental effects. A feasible solution for this issue would be to recycle. The polypropylene samples were submitted to two processes, forced contamination, and recycling, and they were analyzed by solid-phase microextraction gas chromatograph-olfactometry-mass spectrometry. 45 different volatile compounds were identified and 9 of them presented distinct odoriferous activities. Among them, two important markers were detected: diethyl phthalate (probably coming from the catalyst of PP polymerization, intentionally added substance (IAS)), and glycerine (a marker of non-intentionally added substances (NIAS)).

Evaluation of New Antimicrobial Materials Incorporating Ethyl Lauroyl Arginate or Silver into Different Matrices, and Their Safety in Use as Potential Packaging.

A big challenge for today's industry is antimicrobial preservation and the safety of food. An effective solution to this problem can be a modern invention such as antimicrobial packaging. In the presented research the antimicrobial activity of two new active films incorporating silver, as IONPURE IPL, and ethyl lauroyl arginate (LAE) were evaluated, by employing a low-density polyethylene (LDPE) matrix and a biofilm material, respectively. Additionally, LAE was also incorporated into polystyrene (PS) pads by two different methods: by spraying and by immersion of the PS pads into an aqueous LAE solution. LDPE films containing silver did not show any antimicrobial activity against Escherichia coli and Aspergillus flavus, whereas the biofilm containing LAE reduced the growth of Salmonella enterica but did not inhibit Aspergillus flavus. The active PS pads, both sprayed and immersed in LAE solution, also showed antimicrobial activity, causing a reduction of 99.99% of Pseudomonas putida growth. Thermal treatment at 180 °C for 6 and 15 min did not modify the antimicrobial activity of LAE against Salmonella enterica. Moreover, inductively coupled plasma-mass spectrometry (ICP-MS) analysis was performed to check the migration of silver from developed material intended for food packaging applications into food simulant.

Influence of cooking conditions on the migration of silicone oligomers from silicone rubber baking molds to food simulants.

The stability, surface micromorphology, and volatile organic compounds (VOCs) of silicone rubber baking molds (SRBMs) were tested while using the molds under severe conditions: baking at 175 °C, microwaving at 800 W, and freezing at -18 °C. Moreover, migration tests of SRBMs to food simulants (isooctane, 95% ethanol, and Tenax®) at 70 °C for 2 h (accelerated conditions) were performed. The initial total VOCs concentration was 2.53% higher than that recommended by BfR Recommendations on Food Contact Materials. Therefore, the SRBM samples were considered as badly tempered materials, and 18 different types of silicone oligomers were identified during the migration tests. The following percentage of silicone oligomers with a molecular weight lower than 1000 Da in isooctane, 95% ethanol, and Tenax® were detected: 70.7%, 91.8%, and 97.2%, respectively. It has been proven that previous baking treatments effectively reduced the content of silicone oligomers migrating from SRBMs.

Migration of dihydroxyalkylamines from polypropylene coffee capsules to Tenax® and coffee by salt-assisted liquid-liquid extraction and liquid chromatography-mass spectrometry.

Migration of N,N-Bis(2-hydroxyethyl) alkyl(C8-C18)amines from five different polypropylene capsules to Tenax® and coffee powder have been studied. A single step extraction-cleanup procedure using salting out liquid-liquid extraction (SALLE) method followed by ultra-high performance liquid chromatography-tandem mass spectrometry (UHPLC-MS/MS) was applied. The critical parameters on the SALLE procedure such as extracting solvent, extracting volume, sample pH, salt and its concentration were optimized. The recovery values were in the range of 87.5%-106.5%. The %RSD were lower than 3.7%. The limit of detection was improved from 2.3 ng/g in Tenax® to 0.8 ng/g in coffee. The results indicated that the analyzed compounds have the potential to migrate from the polypropylene capsule containers to the coffee. In most of the cases, the migrated values were higher in Tenax® than in coffee in a range between 1.8 and 61%. One sample did not comply with the specific migration limit established by the European Commission.

Analytical Approaches for Analysis of Safety of Modern Food Packaging: A Review.

Nowadays, food packaging is a crucial tool for preserving food quality and has become an inseparable part of our daily life. Strong consumer demand and market trends enforce more advanced and creative forms of food packaging. New packaging development requires safety evaluations that always implicate the application of complex analytical methods. The present work reviews the development and application of new analytical methods for detection of possible food contaminants from the packaging origin on the quality and safety of fresh food. Among food contaminants migrants, set-off migrants from printing inks, polymer degradation products, and aromatic volatile compounds can be found that may compromise the safety and organoleptic properties of food. The list of possible chemical migrants is very wide and includes antioxidants, antimicrobials, intentionally added substances (IAS), non-intentionally added substances (NIAS), monomers, oligomers, and nanoparticles. All this information collected prior to the analysis will influence the type of analyzing samples and molecules (analytes) and therefore the selection of a convenient analytical method. Different analytical strategies will be discussed, including techniques for direct polymer analysis.

Influence of medicinal and aromatic plants into risk assessment of a new bioactive packaging based on polylactic acid (PLA).

A new biodegradable antioxidant active packaging for food applications based on antioxidants from medicinal and aromatic plants incorporated into a polylactic acid matrix was designed and developed. Melt blending processing technique was applied to prepare polylactic acid films loaded by sage and lemon balm leaves. Antioxidant properties of developed active films were investigated using the following methods: 2,2'-azino-bis(3-ethylbenzothiazoline-6-sulphonic acid), 2,2-diphenyl-1-(2,4,6-trinitrophenyl)hydrazyl and a home-made generator of hydroxyl radicals. In addition, reducing power and total phenolic content of polylactic acid films were checked. The results of antioxidant capacity showed that percentage of hydroxylation for active film with lemon balm and sage was 55.5% ±â€¯0.1% and 67.4% ±â€¯0.3%, respectively. The reducing power of active films increased 8 times in comparison to the blank samples. Moreover, extensive investigation of influence of sage and lemon balm leaves on material safety and type of migrants was performed using migration assays. The composition of both non-volatile and volatile compounds of different active packaging films was compared with neat polylactic acid film. Three different food simulants such as 95% (v/v) ethanol, 10% (v/v) ethanol and 3% (w/v) acetic acid were checked. It was shown that the addition of sage and lemon balm leaves into a polylactic acid structure decreased the migration of both linear and cyclic polylactic acid oligomers, currently not legislated by European Union. Besides, total absence or decrease of migration of volatile compounds were observed when using the active films. Both thermal and mechanical properties of films were also evaluated.

Polyamide modified with green tea extract for fresh minced meat active packaging applications.

New antioxidant polyamide was prepared by total immersion in active extract for 48 h. Its antioxidant performance was tested using DPPH (IC50 = 270 ±â€¯21 µg/g) and ORAC (1.52 µmol Trolox/g). In vivo study was done using fresh minced meat stored at 4 °C and analysed after 0, 6, 13, 19, 23 days. After 23 days metmyoglobin (MetMbBK = 31.3 ±â€¯2.9% and MetMbAOX = 25.9 ±â€¯0.8%), CIE L*a*b* (a*BK = 15.0 ±â€¯0.4 and a*AOX = 16.6 ±â€¯0.3) and TBARS (MDABK = 0.0060 ±â€¯0.0003 µg/g and MDAAOX = 0.0044 ±â€¯0.0002 µg/g) showed an improvement of meat shelf life. The results showed that this active film protected meat during 23 days. Migration study to food simulants was done by UPLC®-QqQ-MS and UPLC®-ESI-Q-TOF-MSE. A decrease of oligomers migration (for caprolactam n = 3: BK = 0.050 ±â€¯0.004 mg/Kg; AOX = 0.019 ±â€¯0.001 mg/Kg) was found. INDUSTRIAL RELEVANCE: Nowadays, food industry is focused on improving shelf life of products by controlling its lipid oxidation using natural antioxidants. The obtained results let us design a new active packaging based on natural antioxidants for extending the shelf life of fresh minced meat at industrial scale.

Antioxidant and antimicrobial markers by UPLC®-ESI-Q-TOF-MSE of a new multilayer active packaging based on Arctostaphylos uva-ursi.

49 different non-volatile compounds were determined in Spanish Arctostaphylos uva-ursi leaves using UPLC®-ESI-Q-TOF with MSE technology. Both positive and negative electrospray ionization were applied. MarkerLynx® was proposed as a powerful tool to distinguish samples from eight wild populations of Spain by determining their non-volatile markers. Development of HRMS methods let to analysis of metabolites in plants. Antioxidant and antimicrobial capacities of different extracts were evaluated. Plant extract with the strongest antioxidant and simultaneous good antimicrobial capacity (Lierta) was chosen and incorporated in a multilayer packaging. Then, antioxidant capacity of the new packaging was evaluated and the efficient free radical scavenging properties were demonstrated.

Identification and quantification of odours from oxobiodegradable polyethylene oxidised under a free radical flow by headspace solid-phase microextraction followed by gas chromatography-olfactometry-mass spectrometry.

Recently oxobiodegradable polyethylene gained popularity as food packaging material due to its potential to reduce polymer waste. However, this type of material can release after its oxidation off-odour compounds that affect the organoleptic properties of packaged food. Odour compounds released from both polyethylene and oxobiodegradable polyethylene before and after oxidation under a free radicals flow were investigated after 1 day, 2 days and 3 days of oxidation. The samples were analysed using headspace solid phase microextraction followed by gas chromatography-mass spectrometry and headspace solid phase microextraction coupled to gas chromatography-olfactometry-mass spectrometry. Sixty-two different odorous compounds were identified. 4-methylthio-2-butanone (fruit), nonanal (fat) and 3,6-nonadienal (fat) were present in different materials before oxidation. Multiple headspace-solid phase microextraction has been used to quantify all analytes. The most abundant compound was (Z)-3-hexenyl hexanoate with a concentration range between 1.5791±0.1387µg/g and 4.8181±0.3123µg/g. Compounds such as 2-dodecenal, 2-octenal, 2-pentanol, 3-nonenal, 3,6-nonadienal, ethyl 3-methylbutanoate, ethyl octenoate, hexanone, isopropyl hexanoate, octanal were below their LOD evaluated using MS detector; however, they were detected by gas chromatography-olfactometry. The minimum LOD and LOQ were 0.011µg/g and 0.036µg/g, respectively.

Development and characterisation of HPMC films containing PLA nanoparticles loaded with green tea extract for food packaging applications.

A novel active film material based on hydroxypropyl-methylcellulose (HPMC) containing poly(lactic acid) (PLA) nanoparticles (NPs) loaded with antioxidant (AO) green tea extract (GTE) was successfully developed. The PLA NPs were fabricated using an emulsification-solvent evaporation technique and the sizes were varied to enable a controlled release of the AO from the HPMC matrix. A statistical experimental design was used to optimise the synthesis of the NPs in order to obtain different sizes of nanoparticles and the loading of these into the HPMC matrix was also varied. The physico-chemical properties of the composite films were investigated and the release of the AO was confirmed by migration studies in 50% v/v ethanol/water food simulant. The AO capacity of the GTE released from the active films was studied using the 2,2-diphenyl-1-picrylhydrazyl (DPPH) radical method and the results suggest that the material could potentially be used for extending the shelf-life of food products with high fat content.