Subterranean transport of microplastics as evidenced in karst springs and their characterization using Raman spectroscopy.

The increasing use of plastic materials has led to accumulation of large amounts of plastic waste in environment and a global challenge to be tackled with. The natural process of macro-plastics aging generates a multitude of secondary microplastic fragments accumulating in all areas of the planet. The pollution with microplastics of large water bodies, such as rivers, seas and oceans was already proven, but the presence of microplastics even in karst spring water was not reported yet. In this study, Raman micro-spectroscopy was used to confirm the presence of microplastics in the spring water samples collected from two rural karst springs in the Apuseni Mountains (Țarina and Josani), North-Western Romania. Two sets of water samples of 1000 L collected in spring time 2021 and one in autumn 2021 were filtered and analyzed. Using the Python programming language and combining two separate Raman databases, one for plastics and the other for pigments, we established a customized database to unambiguously identify the type of plastic and pigment present in the discovered micro-fragments. The generated reference pigment-plastic spectra were compared to those of potential microplastics found on filters and Pearson's coefficient was used to measure the level of similarity. The presence of microplastics in karst spring waters was confirmed and a quantitative estimation expressed as number of fragments or fibers per liter was 0.034 in Josani and 0.06 in Țarina karst spring. Five months later sampling (autumn 2021) revealed 0.05 microplastics per liter. The spectral results revealed that most microplastics found were dominated by polyethylene terephthalate (PET), followed by polypropylene and interestingly, abundant blue micro-fragments were identified according to their copper phthalocyanine pigments (pigment Blue 15) or indigo carmine (pigment Blue 63) characteristic spectral fingerprints, which surpassed the inherent spectral background level characteristic for the Raman spectra of naturally contaminated waste micro-samples. Their origin in mountain karst spring waters and potential decrease in time is discussed.

pH-Dependent Behavior of Novel 5-FU Delivery System in Environmental Conditions Comparable to the Gastro-Intestinal Tract.

A biogenic carrier for 5-fluorouracil (5-FU) loading and subsequent tableting as a new drug formulation for slow release has been proposed using the biomineral from blue crab carapace. Due to its highly ordered 3D porous nanoarchitecture, the biogenic carbonate carrier could achieve increased effectiveness in colorectal cancer cure provided that the formulation would successfully pass through the gastric acid conditions. Following the recently proven viability of the concept by demonstrating the slow release of the drug from the carrier using the highly sensitive SERS technique, here we investigated the 5-FU release from the composite tablet drug in pH conditions replicating the gastric environment. The released drug from the tablet was studied in solutions with three relevant pH values, pH 2, pH 3, and pH 4. The 5-FU SERS spectral signature for each pH value was used to build calibration curves for quantitative SERS analysis. The results suggested a similarly slow-releasing pattern in acid pH environments to that in neutral conditions. Although biogenic calcite dissolution was expected in acid conditions, the X-ray diffraction and Raman spectroscopy showed preservation of calcite mineral along with the monohydrocalcite during acid solution exposure for two hours. The total released amount in a time course of seven hours, however, was lower in acidic pH solutions, with a maximum fraction of ~40% of the total amount of loaded drug, for pH 2, as opposed to ~80% for neutral values. Nonetheless, these results clearly prove that the novel composite drug retains its slow-releasing character in environmental conditions compatible with the gastrointestinal pH and that it is a viable and biocompatible alternative for oral delivery of anticancer drug to reach the lower gastro-intestinal tract.

Pilot SERS Monitoring Study of Two Natural Hypersaline Lake Waters from a Balneary Resort during Winter-Months Period.

Water samples from two naturally hypersaline lakes, renowned for their balneotherapeutic properties, were investigated through a pilot SERS monitoring program. Nanotechnology-based techniques were employed to periodically measure the ultra-sensitive SERS molecular characteristics of the raw water-bearing microbial community and the inorganic content. Employing the Pearson correlation coefficient revealed a robust linear relationship between electrical conductivity and pH and Raman and SERS spectral data of water samples, highlighting the interplay complexity of Raman/SERS signals and physicochemical parameters within each lake. The SERS data obtained from raw waters with AgNPs exhibited a dominant, reproducible SERS feature resembling adsorbed ß-carotene at submicromole concentration, which could be related to the cyanobacteria-AgNPs interface and supported by TEM analyses. Notably, spurious SERS sampling cases showed molecular traces attributed to additional metabolites, suggesting multiplexed SERS signatures. The conducted PCA demonstrated observable differences in the ß-carotene SERS band intensities between the two lakes, signifying potential variations in picoplankton abundance and composition or environmental influences. Moreover, the study examined variations in the SERS intensity ratio I245/I1512, related to the balance between inorganic (Cl--induced AgNPs aggregation) and organic (cyanobacteria population) balance, in correlation with the electrical conductivity. These findings signify the potential of SERS data for monitoring variations in microorganism concentration, clearly dependent on ion concentration and nutrient dynamics in raw, hypersaline water bodies.

Detection and Characterization of Nodularin by Using Label-Free Surface-Enhanced Spectroscopic Techniques.

Nodularin (NOD) is a potent toxin produced by Nodularia spumigena cyanobacteria. Usually, NOD co-exists with other microcystins in environmental waters, a class of cyanotoxins secreted by certain cyanobacteria species, which makes identification difficult in the case of mixed toxins. Herein we report a complete theoretical DFT-vibrational Raman characterization of NOD along with the experimental drop-coating deposition Raman (DCDR) technique. In addition, we used the vibrational characterization to probe SERS analysis of NOD using colloidal silver nanoparticles (AgNPs), commercial nanopatterned substrates with periodic inverted pyramids (KlariteTM substrate), hydrophobic Tienta® SpecTrimTM slides, and in-house fabricated periodic nanotrenches by nanoimprint lithography (NIL). The 532 nm excitation source provided more well-defined bands even at LOD levels, as well as the best performance in terms of SERS intensity. This was reflected by the results obtained with the KlariteTM substrate and the silver-based colloidal system, which were the most promising detection approaches, providing the lowest limits of detection. A detection limit of 8.4 × 10-8 M was achieved for NOD in solution by using AgNPs. Theoretical computation of the complex vibrational modes of NOD was used for the first time to unambiguously assign all the specific vibrational Raman bands.

Rutin bioconjugates as potential nutraceutical prodrugs: An in vitro and in ovo toxicological screening.

Rutin (RUT) is considered one the most attractive flavonoids from a therapeutic perspective due to its multispectral pharmacological activities including antiradical, anti-inflammatory, antiproliferative, and antimetastatic among others. Still, this compound presents a low bioavailability what narrows its clinical applications. To overcome this inconvenience, the current paper was focused on the synthesis, characterization, and toxicological assessment of two RUT bioconjugates obtained by enzymatic esterification with oleic acid (OA) and linoleic acid (LA)-rutin oleate (RUT-O) and rutin linoleate (RUT-L), as flavonoid precursors with improved physicochemical and biological properties. Following the enzymatic synthesis in the presence of Novozyme® 435, the two bioconjugates were obtained, their formation being confirmed by RAMAN and FT-IR spectroscopy. The in vitro and in ovo toxicological assessment of RUT bioconjugates (1-100 µM) was performed using 2D consecrated cell lines (cardiomyoblasts - H9c2(2-1), hepatocytes-HepaRG, and keratinocytes-HaCaT), 3D reconstructed human epidermis tissue (EpiDerm™), and chick chorioallantoic membranes, respectively. The results obtained were test compound, concentration-and cell-type dependent, as follows: RUT-O reduced the viability of H9c2(2-1), HepaRG, and HaCaT cells at 100 µM (to 77.53%, 83.17%, and 78.32%, respectively), and induced cell rounding and floating, as well as apoptotic-like features in the nuclei of all cell lines, whereas RUT-L exerted no signs of cytotoxicity in all cell lines in terms of cell viability, morphology, and nuclear integrity. Both RUT esters impaired the migration of HepaRG cells (at 25 µM) and lack irritative potential (at 100 µM) in vitro (tissue viability >50%) and in ovo (irritation scores of 0.70 for RUT-O, and 0.49 for RUT-L, respectively). Computational predictions revealed an increased lipophilicity, and reduced solubility, drug-likeness and drug score of RUT-O and RUT-L compared to their parent compounds-RUT, OA, and LA. In conclusion, we report a favorable toxicological profile for RUT-L, while RUT-O is dosage-limited since at high concentrations were noticed cytotoxic effects.

Comparative screening the life-time composition and crystallinity variation in gilthead seabream otoliths Sparus aurata from different marine environments.

Differences in crystallinity, structure and composition variation along the growing direction in gilthead seabream, Sparus aurata otoliths that inhabited different environments were determined to evaluate the correlation of spectroscopic and chemical data with the lifetime development and movement pattern. The Raman spectroscopy signal provided the characteristic bands whose Full Width at Half Maximum (FWHM) were used to track the signal variability. The FWHM showed an initial increase in the core area, followed by a decrease depicting two minima coinciding growth rings. The crystal discontinuity linked to annual rings was confirmed. The FWHM pattern followed cycle in the individual's activity. However, no significant correlation with FWHM and environmental factors although the slope of the FWHM variation distinguished aquaculture and costal groups from open sea and transitional, estuarine waters. Raman data were further correlated with morphological and elemental composition obtained via SEM-EDX and by LA-ICP-MS. SEM clearly confirmed CRM findings. Finally, multiparameter analysis of Ba/Ca concentrations obtained by LA-ICP-MS indicated the separation of groups associated with aquaculture and transitional waters due lowest variability in the elemental composition. Other groups are more variable possibly due to the water oligotrophic character and greater variability in prey availability in each environment. Results of the present study showed the additional potential of Raman spectroscopy as a complementary tool for inference of migration or origin of fish based on otolith composition and structure like other well-established technique.

Raman technology application for plastic waste management aligned with FAIR principle to support the forthcoming plastic and environment initiatives.

Plastic production and worldwide use of plastic materials have continued to rise due to their convenience and excellent marketing advantages. This is generating an environmental crisis and global scale pollution which is one of the greatest threats to our planet. One of the best responses could be accomplished by improving recycling and waste management strategies. In this paper we conducted Raman analyses of representative stock of plastics aged in terrestrial or aquatic environments spanning in age up to 15 years. We aimed to establish any potential influence of the aging conditions on the Raman signature of specific plastics. This information is further used to build up a Raman logic gate for automatic sorting of plastic waste recovered from environment. Pigments and aging introduced indeed small changes in the Raman signature of the respective plastics. However, we were able to identify unique spectral ranges characteristic for the main plastic types and intensity threshold of fingerprint bands sufficiently strong for building robust Raman barcodes for sorting. Waste plastics Raman data handling and the proposed methodology for sorting complies with the FAIR (Findability, Accessibility, Interoperability and Reusability) principles of scientific data, being useful for researchers, policymakers and stakeholders. Our spectral characterization of solid plastic waste comes in support of improved waste plastic management and could have economic and environmental positive impact.

Wasted Biomaterials from Crustaceans as a Compliant Natural Product Regarding Microbiological, Antibacterial Properties and Heavy Metal Content for Reuse in Blue Bioeconomy: A Preliminary Study.

The compliance of crab shells traditionally used as a complex natural product for agricultural soil amendment with modern biofertilizers' quality and safety requirements was investigated. Shells waste from the Blue crab, Callinectes sapidus and the Green crab, Carcinus aestuarii were tested for macronutrients, heavy metals, bacteria content, and antimicrobial properties. Such information is crucial for further utilization of the biogenic powders for any composite formulation in added-value by-products. The calcium carbonate-rich hard tissue yield was 52.13% ± 0.015 (mean ± S.D.) and 64.71% ± 0.144 from the blue and green crabs, respectively. The contents of Pb, Ni, Zn, Cr (VI), and Cu were several orders of magnitude below the prescribed limit by EU biofertilizer legislation, with Fe, Mn (not prescribed), and As being the most abundant. The content of As and Cd from the material considered here was within limits. The shells contain no colony-forming units of Salmonella spp. and compliant levels of Escherichia coli; moreover, the shell micro-powder showed dose-dependent growth inhibition of Pseudomonas aeruginosa and Staphylococcus aureus. In summary, the waste crab shells present a complex natural product as plant biofertilizer following the circular economy concepts.

Development and Characterization of Fe3O4@Carbon Nanoparticles and Their Biological Screening Related to Oral Administration.

The current study presents the effect of naked Fe3O4@Carbon nanoparticles obtained by the combustion method on primary human gingival fibroblasts (HGFs) and primary gingival keratinocytes (PGKs)-relevant cell lines of buccal oral mucosa. In this regard, the objectives of this study were as follows: (i) development via combustion method and characterization of nanosized magnetite particles with carbon on their surface, (ii) biocompatibility assessment of the obtained magnetic nanoparticles on HGF and PGK cell lines and (iii) evaluation of possible irritative reaction of Fe3O4@Carbon nanoparticles on the highly vascularized chorioallantoic membrane of a chick embryo. Physicochemical properties of Fe3O4@Carbon nanoparticles were characterized in terms of phase composition, chemical structure, and polymorphic and molecular interactions of the chemical bonds within the nanomaterial, magnetic measurements, ultrastructure, morphology, and elemental composition. The X-ray diffraction analysis revealed the formation of magnetite as phase pure without any other secondary phases, and Raman spectroscopy exhibit that the pre-formed magnetic nanoparticles were covered with carbon film, resulting from the synthesis method employed. Scanning electron microscopy shown that nanoparticles obtained were uniformly distributed, with a nearly spherical shape with sizes at the nanometric level; iron, oxygen, and carbon were the only elements detected. While biological screening of Fe3O4@Carbon nanoparticles revealed no significant cytotoxic potential on the HGF and PGK cell lines, a slight sign of irritation was observed on a limited area on the chorioallantoic membrane of the chick embryo.

Biocompatible Magnetic Colloidal Suspension Used as a Tool for Localized Hyperthermia in Human Breast Adenocarcinoma Cells: Physicochemical Analysis and Complex In Vitro Biological Profile.

Magnetic iron oxide nanoparticles are the most desired nanomaterials for biomedical applications due to their unique physiochemical properties. A facile single-step process for the preparation of a highly stable and biocompatible magnetic colloidal suspension based on citric-acid-coated magnetic iron oxide nanoparticles used as an effective heating source for the hyperthermia treatment of cancer cells is presented. The physicochemical analysis revealed that the magnetic colloidal suspension had a z-average diameter of 72.7 nm at 25 °C with a polydispersity index of 0.179 and a zeta potential of -45.0 mV, superparamagnetic features, and a heating capacity that was quantified by an intrinsic loss power analysis. Raman spectroscopy showed the presence of magnetite and confirmed the presence of citric acid on the surfaces of the magnetic iron oxide nanoparticles. The biological results showed that breast adenocarcinoma cells (MDA-MB-231) were significantly affected after exposure to the magnetic colloidal suspension with a concentration of 30 µg/mL 24 h post-treatment under hyperthermic conditions, while the nontumorigenic (MCF-10A) cells exhibited a viability above 90% under the same thermal setup. Thus, the biological data obtained in the present study clearly endorse the need for further investigations to establish the clinical biological potential of synthesized magnetic colloidal suspension for magnetically triggered hyperthermia.

Single-cell Raman micro-spectroscopy for tracking of carotenoids in cyanobacteria exposed to Mn and Zn doped ferrite nanoparticles.

Ferrite nanoparticles are increasingly produced and exploited as adsorbents for environmental pollutants. However, their impact on the aquatic microbiota such as cyanobacteria, are not yet investigated. Targeting the environmental monitoring context, in this paper we explored for the first time if any change in the carotenoid signal from cyanobacteria Coelomoron pussilum (AICB 1012) exposed to non-lethal doses of Mn and Zn doped ferrite nanoparticles (NPs) can be associated with the nano-aggression on single-cell level, using micro-Raman spectroscopy. UV-Vis absorption spectroscopy of the bulk culture and single-cell Raman microscopy showed that the carotenoid signal increases relative to the chlorophyll contribution upon exposure of the cells to the Mn-ferrite NPs throughout the 7 days of the experiment. The red-shift and broadening of the strongest carotenoid Raman band arising from (CC) stretching modes indicates the change of carotenoid profile towards increased amount of ß-carotene in answer to the NPs stress. The increase of this band intensity relative to the fluorescence background was also observed in Zn-ferrite NPs treatment. Using a simplified and rapid sample preparation procedure, electron microscopy in both transmission and scanning modes, showed greater coverage of the cells by the stable colloidal AgNPs than by the magnetic ferrite NPs. The latter mostly clumped together rather than adhering to the cells. The combined single-cell micro-Raman tracking of physiological response of the unicellular photosynthetic microorganisms coupled with electron microscopy approach to visualise cell-NPs interaction and the extracellular polymeric substance secretion holds the promise for rapid assessment of the NPs-induced environmental stress acting on the unicellular organisms.

Testing the limits of FT-Raman spectroscopy for wine authentication: Cultivar, geographical origin, vintage and terroir effect influence.

FT-Raman spectroscopy represents an environmentally friendly technique, suitable for the analysis of high-water content food matrices, like wines, due to its relatively weak water bending mode in the fingerprint region. Based on metabolomics applied to FT-Raman spectra, this study presents the classifications achieved for a sample set comprising 126 wines, originated from Romania and France, with respect to cultivar, geographical origin and vintage. Cultivar recognition was successfully performed among four varieties (Sauvignon, Riesling, Chardonnay, Pinot Gris) while subtle particularities exiting between the Chardonnay wines, coming from the two countries, because of terroir influences were pointed out. The obtained separations of 100% in both initial and cross-validation procedure for geographical differentiation between the two origin countries, as well as, among the three Romanian areas (Transylvania, Muntenia and Moldova) were also discussed. Apart of this, the limitations and the importance of choosing a meaningful data set, in terms of representativity for each classification criterion, are addressed in the present work.

Microsphere packages of carotenoids: intact sea urchin eggs tracked by Raman spectroscopy tools.

Although natural exposure to ambient UV radiation in oligotrophic seawater at small depths can reach the levels responsible for cellular damage, the sea urchin Paracentrotus lividus is frequently in such sites, particularly on the southern Adriatic Sea shore. Spawning their eggs and spending their early life stage in rocky shores at depths of 0.5-2 m are the results of their successful adaptation strategies, although adults may dwell at greater depths. Surprisingly, there is a paucity of reports regarding the carotenoid content in sea urchin eggs. Beyond their important role in photoprotection against high UV exposure, cell division and early development, the content and distribution of carotenoids contribute to the successful survival of sea urchins and also determine the color of their gonads (roe), which is of commercial importance as a delicacy. Herein, for the first time, we have described the carotenoid content and distribution in intact, freshly released eggs of P. lividus species, non-destructively employing resonance Raman spectroscopy and imaging; near-infrared Raman spectroscopy revealed additional molecular carotenoid content. Echinenone and ß-carotene resonance Raman signals were the most intense, and they were identified as the principal carotenoids that are preferentially accumulated in eggs rather than in gonads. Raman imaging in confocal mode revealed the uniform distribution of the carotenoid signal over the whole eggs, while the distribution of proteins appeared spotted. Egg carotenoids generally maintained their identity after 2 months of dry storage, with slight signs of C[double bond, length as m-dash]C bond oxidation. The potential utilization of P. lividus sea urchin eggs as valuable microsphere packages of native carotenoids is discussed.

Citrus fruits freshness assessment using Raman spectroscopy.

The freshness of citrus fruits commonly available in the market was non-destructively assessed by Raman spectroscopy. Intact clementine, mandarin and tangerine species were characterised concerning their carotenoids skin Raman signalling in a time course from the moment they were acquired as fresh stock, supplying the market, to the physical degradation, when they were no longer attractive to consumers. The freshness was found to strongly correlate to the peel Raman signal collected from the same area of the intact fruits in a time course of a maximum of 20days. We have shown that the intensity of the carotenoid Raman signal is indeed a good indicator of fruit freshness and introduced a Raman coefficient of freshness (CFresh), whose time course is linearly decreasing, with different slope for different citrus groups. Additionally, we demonstrated that the freshness assessment could be achieved using a portable Raman instrument. The results could have a strong impact for consumer satisfaction and the food industry.

Amnesic shellfish poisoning biotoxin detection in seawater using pure or amino-functionalized Ag nanoparticles and SERS.

Domoic acid (DA) biotoxin responsible for the amnesic shellfish poisoning (ASP) has been unambiguously detected in seawater in a broad range of concentration, with both pure and amino-functionalized Ag nanoparticles employed for surface enhanced Raman scattering (SERS). To achieve this, a comprehensive SERS study on DA dissolved in distilled water has been conducted. SERS of DA dissolved in seawater in concentrations ranging from 3.3 × 10(-4) to 3.3 × 10(-8) mol l(-1) exhibited specific signal, completely different to those of the corresponding DA aqueous solutions, due to the seawater interference in the overall SERS effect. In order to assess the capability of the technique as a cheaper alternative for rapid and unambiguous detection of the DA biotoxin in seawater, three detection schemes have been proposed. DA was detectable at 0.33 nmoll(-1) concentration (0.33) dissolved in distilled water and 0.033 nmol l(-1) (0.033 ppb) in seawater respectively, much lower than the admitted level by the current regulation. A solvent specific interaction of DA with the NPs was concluded, since DA aqueous solution added to Ag nanoparticles provided different SERS signal compared to that of DA directly dissolved in seawater. Employing amino-functionalized Ag nanoparticles with 4-aminothiophenol as SERS tag, SERS signal of DA on amino-AgNPs revealed significant specificity associated with the aromatic primary amine interaction of the SERS tag with DA, thus allowing DA detection in seawater at 4.16 × 10(-4) mol l(-1) concentration, much higher than in the case of pure NPs. To highlight the findings, a brief literature review to date on the DA biotoxin detection was also provided.

Molecular conformation changes along the malignancy revealed by optical nanosensors.

An interdisciplinary approach employing functionalized nanoparticles and ultrasensitive spectroscopic techniques is reported here to track the molecular changes in early stage of malignancy. Melanoma tissue tracking at molecular level using both labelled and unlabelled silver and gold nanoparticles has been achieved using surface enhanced Raman scattering (SERS) technique. We used skin tissue from ex vivo mice with induced melanoma. Raman and SERS molecular characterization of melanoma tissue is proposed here for the first time. Optical nanosensors based on Ag and Au nanoparticles with chemisorbed cresyl violet molecular species as labels revealed sensitive capability to tissues tagging and local molecular characterization. Sensitive information originating from surrounding native biological molecules is provided by the tissue SERS spectra obtained either with visible or NIR laser line. Labelled nanoparticles introduced systematic differences in tissue response compared with unlabelled ones, suggesting that the label functional groups tag specific tissue components revealed by proteins or nucleic acids bands. Vibrational data collected from tissue are presented in conjunction with the immunohistochemical analysis. The results obtained here open perspectives in applied plasmonic nanoparticles and SERS for the early cancer diagnostic based on the appropriate spectral databank.

Study of the betulin enriched birch bark extracts effects on human carcinoma cells and ear inflammation.

BACKGROUND: Pentacyclic triterpenes, mainly betulin and betulinic acid, are valuable anticancer agents found in the bark of birch tree. This study evaluates birch bark extracts for the active principles composition. RESULTS: New improved extraction methods were applied on the bark of Betula pendula in order to reach the maximum content in active principles. Extracts were analyzed by HPLC-MS, Raman, SERS and 13C NMR spectroscopy which revealed a very high yield of betulin (over 90%). Growth inhibiting effects were measured in vitro on four malignant human cell lines: A431 (skin epidermoid carcinoma), A2780 (ovarian carcinoma), HeLa (cervix adenocarcinoma) and MCF7 (breast adenocarcinoma), by means of MTT assay. All of the prepared bark extracts exerted a pronounced antiproliferative effect against human cancer cell lines. In vivo studies involved the anti-inflammatory effect of birch extracts on TPA-induced model of inflammation in mice. CONCLUSIONS: The research revealed the efficacy of the extraction procedures as well as the antiproliferative and anti-inflammatory effects of birch extracts.

Evaluation and differentiation of the Betulaceae birch bark species and their bioactive triterpene content using analytical FT-vibrational spectroscopy and GC-MS.

BACKGROUND: Aiming to obtain the highest triterpene content in the extraction products, nine bark samples from the forest abundant flora of Apuseni Mountains, Romania were Raman spectroscopically evaluated. Three different natural extracts from Betula pendula Roth birch bark have been obtained and characterized using Fourier transform vibrational spectra. RESULTS: This study shows that principal components of the birch tree extract can be rapidly recognized and differentiated based on their vibrational fingerprint band shape and intensity. The vibrational spectroscopy results are supported by the GC-MS data. Based on IR and Raman analysis, one can conclude that all the extracts, independent on the solvent(s) used, revealed dominant betulin species, followed by lupeol. CONCLUSIONS: Since Raman measurements could also be performed on fresh plant material, we demonstrated the possibility to apply the present results for the prediction of the highest triterpene content in bark species, for the selection of harvesting time or individual genotypes directly in the field, with appropriate portable Raman equipment.

Raman, surface enhanced Raman spectroscopy, and DFT calculations: a powerful approach for the identification and characterization of 5-fluorouracil anticarcinogenic drug species.

The normal Raman and SERS spectra of 5-fluorouracil (5-FU) in water solution and attached to a biological artificial model (a silver colloid) at different pH values were recorded and discussed. The DFT calculation results helped us to establish for the first time the most stable resonance structure for each of the tautomeric forms (i.e., two enol and two enolate forms) and to interpret the Raman and SERS spectra. At alkaline pH, both deprotonated forms of 5-FU were found to be present in solution and to adsorb on the Ag surface in a perpendicular orientation or an orientation not significantly tilted from the surface normal. The N3-deprotonated form seems to be the dominant tautomer in the adsorbed state, more probably attached through the O7 atom. At acid pH values, the N3-deprotonated form was again found to be the mainly chemisorbed species adopting a similar orientation. The combination of these two approaches (i.e., the theoretical and experimental one) proved to be a viable candidate for inclusion in a rapid, sensitive biological method of detecting and studying such essential anticarcinogenic species or biological threats in different conditions.