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Stateva, Roumiana

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  • Novel insights into the biological activity of Croton lechleri twigs extracts and advancements in their sustainable recovery
    Publication . Tzintzarov, Alexander; Boyadzhieva, Stanislava S.; Coelho, Jose; Tsvetanova, Flora; Petrova, Maria; Stoev, Georgi; Yankov, Dragomir; Ugrinova, Iva; Stateva, Roumiana
    Sangre de drago, the sap of Croton lechleri Müll. Arg. tree, has been used for centuries in traditional medicine owing to its diverse biological activities. Extracts derived from different parts of the species contain a multitude of phytochemicals with varied applications. Twigs, however, are among the least studied parts of the plant. This study unveils new biological activities of Croton lechleri twig extracts recovered by applying Soxhlet and advanced green techniques. For all extracts, total phenolic content and antioxidant activity were determined. Subsequently, four were selected, and their cytotoxic effects were assessed on both normal (HaCat) and malignant melanoma (A375) skin cell lines using the MTT assay and trypan blue exclusion assay. All showed dose-dependent cytotoxicity, with the Soxhlet ethanol extract demonstrating the highest selectivity towards A375 cells over HaCat cells. The extracts induced apoptosis and necrosis, as confirmed by Annexin V/PI dual-labeling and flow cytometry, highlighting their ability to trigger programmed cell death in cancer cells. The selective inhibition of cell cycle progression in A375 compared to HaCat observed both for Soxhlet ethanol and pressurized ethanol extracts induces cell cycle arrest at multiple points, primarily in the G1 and G2/M phases, and significantly reduces DNA synthesis as evidenced by the decrease in the S-phase population, confirmed by the EdU assay. Consequently, the Soxhlet extract composition was analyzed using LC-MS, which revealed their richness in polyphenolic compounds, particularly flavonoids from the flavonol subclass.
  • Sustainable transformation of two algal species of different genera to high-value chemicals and bioproducts
    Publication . Tsvetanova, Flora; Boyadzhieva, Stanislava S.; Coelho, Jose; Yankov, Dragomir; Stateva, Roumiana
    This study investigates the potential of two algae species from different genera, namely the recently isolated Scenedesmus obliquus BGP and Porphyridium cruentum, from the perspective of their integral sustainable transformation to valuable substances. Conventional Soxhlet and environmentally friendly supercritical fluid extraction were applied to recover oils from the species. The extracts were characterized through analytical techniques, such as GC-Fid and LC-MS/MS, which allowed their qualitative and quantitative differentiation. Thus, P. cruentum oils contained up to 43% C20:4 and C20:5 fatty acids, while those of S. obliquus BGP had only residual amounts. The LC-MS/MS analysis of phenolic compounds in the S. obliquus BGP and P. cruentum extracts showed higher content of 3-OH-4-methoxybenzoic acid and kaempferol 3-O-glycoside in the former and higher amounts of ferulic acid in the latter. Total phenolic content and antioxidant activity of the oils were also determined and compared. The compositional analysis of the oil extracts revealed significant differences and varying potentialities based on their genera and method of extraction. To the best of our knowledge our work is unique in providing such detailed information about the transformation prospects of the two algae species to high-value chemicals and bioproducts.
  • Assessment of gnaphalium viscosum (Kunth) valorization prospects: sustainable recovery of antioxidants by different techniques
    Publication . Boyadzhieva, Stanislava; Coelho, Jose; Errico, Massimiliano; Reynel-Avilla, H. Elizabeth; Yankov, Dragomir S.; Bonilla-Petriciolet, A.; Stateva, Roumiana
    This work investigates the prospects for exploitation of Gnaphalium viscosum (Kunth) abundant but with limited applications till present biomass. The feasibility of traditional techniques (two-phase solvent, and the benchmark Soxhlet extraction) and supercritical extraction without/with a cosolvent at T = 40–60 ◦C and p = 30–50 MPa was examined to explore the possibility of recovering phytochemicals from G. viscosum leaves, flowers and stems. The efficiency of the techniques was assessed and compared based on yield, influence of solvents used, total phenolic content and antioxidant activity of the extracts. Phenolics of different complexities were identified and quantified by applying LC (LC–MS/MS, and LC–HRAM), while the fatty acid profile was determined by GC– FID. The results of this extensive study demonstrated the huge valorization potential and prospects of G. viscosum, since highly potent antioxidants such as kaempferol-3-O-β-d-glucoside (astragalin), and chlorogenic acid were ascertained in considerable amounts. Furthermore, for the first time, the presence of leontopodic acid, a greatly substituted derivative of glucaric acid, was detected in the species.
  • Brewer's spent grain, coffee grounds, burdock, and willow-four examples of biowaste and biomass valorization through advanced green extraction technologies
    Publication . Errico, Massimiliano; Coelho, Jose; Stateva, Roumiana; Christensen, Knud Villy; El-Houri, Rime Bahij; Tronci, Stefania
    This paper explores the transformation of biowastes from food industry and agriculture into high-value products through four examples. The objective is to provide insight into the principles of green transition and a circular economy. The first two case studies focus on the waste generated from the production of widely consumed food items, such as beer and coffee, while the other two examine the potential of underutilized plants, such as burdock and willow, as sources of valuable compounds. Phenolic compounds are the main target in the case of brewer's spent grain, with p-coumaric acid and ferulic acid being the most common. Lipids are a possible target in the case of spent coffee grounds with palmitic (C16:0) and linoleic (C18:2) acid being the major fatty acids among those recovered. In the case of burdock, different targets are reported based on which part of the plant is used. Extracts rich in linoleic and oleic acids are expected from the seeds, while the roots extracts are rich in sugars, phenolic acids such as chlorogenic, caffeic, o-coumaric, syringic, cinnamic, gentisitic, etc. acids, and, interestingly, the high-value compound epicatechin gallate. Willow is well known for being rich in salicin, but picein, (+)-catechin, triandrin, glucose, and fructose are also obtained from the extracts. The study thoroughly analyzes different extraction methods, with a particular emphasis on cutting-edge green technologies. The goal is to promote the sustainable utilization of biowaste and support the green transition to a more environmentally conscious economy.
  • Supercritical CO2 extraction of spent coffee grounds. Influence of co-solvents and characterization of the extracts
    Publication . Coelho, Jose; Filipe, Rui; Robalo, M. Paula; Boyadzhieva, Stanislava; Cholakov, Georgi; Stateva, Roumiana
    Spent coffee grounds (SCGs) were extracted with supercritical CO2 at temperatures of 313 K and 333 K, pressures up to 50.0 MPa and different scCO(2) flow rates. The addition of co-solvents isopropanol, ethanol and ethyl lactate decreased the time to achieve the maximum oil yield, to half of that with pure CO2. Analysis of the extracted oils by NMR, showed that caffeine content was (0.56-3.96) % and DUFA and MUFA contents changed within (39.8-42.0) % and (12.9-15.8) %, respectively. The analyses of fatty acids performed by GC-FID revealed the composition of the oils being 76 % of the total constituted by palmitic and linoleic esters. The DPPH antioxidant capacity was improved up to 12.5 times with co-solvents. The kinetics of oil extraction from SCGs with pure scCO(2) was modeled successfully by combining for the first time representation of the oil mixture by a single virtual molecule with group contribution methods.
  • Microwave-assisted extraction of phenolic compounds from spent coffee grounds. Process optimization applying design of experiments
    Publication . Coelho, Jose; Robalo, Maria Paula; Boyadzhieva, Stanislava; Stateva, Roumiana
    In this study, sustainable technology microwave-assisted extraction (MAE) in association with green solvents was applied to recover phenolic compounds from spent coffee grounds (SCGs). A design of experiments (DOE) was used for process optimization. Initially, a 2(4-1) two level Fractional Factorial Design was used and ratios "solvent to solute" and "ethanol to water" were identified as the significant experimental factors. Consequently, Central Composite Design (CCD) was applied to analyze the effects of the significant variables on the response yield, total polyphenols content (TPC), and antioxidant activity (AA) by the DPPH assay method, and quadratic surfaces to optimize those responses were generated. The values of the significant factors of 16.7 (solvent/solute) and 68.9% (ethanol/water) were optimized simultaneously the yield (%) at 6.98 +/- 0.27, TPC (mg GAE/g) at 117.7 +/- 6.1, and AA (mu mol TE/g) at 143.8 +/- 8.6 and were in excellent agreement with those predicted from the CCD model. The variations of the compositions of the lipids, caffeine, pentacyclic diterpenes, and FAME as a function of the dominant factor % ethanol in the solvent mixture were analyzed by applying NMR and GC-FID, and the results obtained confirmed their determinative significance.
  • Optimization of oil recovery from Japonica luna rice bran by supercritical carbon dioxide applying design of experiments: characterization of the oil and mass transfer modeling
    Publication . Coelho, Jose; Robalo, Maria Paula; Fernandes, Inês S.; Stateva, Roumiana
    This study presents an optimization strategy for recovery of oil from Japonica Luna rice bran using supercritical carbon dioxide (scCO2), based on design of experiments (DoE). Initially, a 24−1 two level fractional factorial design (FFD) was used, and pressure, temperature, and scCO2 flow rate were determined as the significant variables; while the yield, total flavonoids content (TFC), and total polyphenols content (TPC) were the response functions used to analyze the quality of the extracts recovered. Subsequently, central composite design (CCD) was applied to examine the effects of the significant variables on the responses and create quadratic surfaces that optimize the latter. The following values of pressure = 34.35 MPa, temperature = 339.5 K, and scCO2 flow rate = 1.8 × 10−3 kg/min were found to simultaneously optimize the yield (6.83%), TPC (61.28 μmol GAE/g ext), and TFC (1696.8 μmol EC/g ext). The fatty acid profile of the oils was characterized by GC-FID. It was demonstrated that the acids in largest quantities are C16:0 (15–16%), C18:1 (41%), and C18:2 (38–39%). Finally, three mass transfer models were applied to determine the mass transfer coefficients and assess the cumulative extraction curves, with an AAD% of 4.16, for the best model.
  • Biomass and waste conversion and valorization to chemicals, energy and fuels
    Publication . Coelho, Jose; Stateva, Roumiana
    Biomass and biowaste are a storehouse of a wide variety of valuable compounds, and their conversion to energy and bioproducts, platform chemicals, biofuels, etc., offer a multifaceted approach to addressing some of the most pressing global challenges, including climate change, waste management, and sustainable development. Efficient and viable biomass valorization, however, requires the introduction of innovative advanced techniques and processes with limited applications of conventional organic solvents. Techniques using green pressurized fluids as solvents and processes like chemical catalysis, biochemical and thermochemical conversion, enzymatic hydrolysis, fermentation, anaerobic digestion, etc. can transform the biomass into bioactive compounds, bio-oils, and other valuable products. The latter can subsequently be used for the production of pharmaceuticals, food additives and supplements, platform chemicals, biofuels, or materials like biochar, which can find applications as a soil amendment or as a carbon sequestration tool. Embracing such technologies epitomizes the new synergistic approach for the development of holistic zero-waste biorefineries, which are at the nexus between sustainability and circular bioeconomics and will not only reduce environmental impact but also pave the way to a cleaner and more economically resilient future.