Enriched Extract Research Articles

Unveiling the anti-proliferative and pro-thermogenic activity of Staurosirella pinnata (Bacillariophyta) bioproducts

Biorefinery co-products obtained from biomass of the field isolated diatom Staurosirella pinnata have been investigated for their bioactivity on selected human and murine cell lines. Biomass grown in indoor photobioreactor, collected at the stationary phase, was subjected to methanolic extraction to produce a crude extract, sequentially fractionated (hydrophilic-hydrophobic gradient) to obtain six fractions (A4, A5, A7; B4, B5, B6). Fractions' bioactivity was tested on BRAFwt and BRAFV600E human melanoma cell lines, compared to normal keratinocytes. All fractions showed cytotoxic activity, but only fraction A4 showed a remarkable cell-type selectivity, with cell death induction up to 70 % on melanoma cells. Further characterization allowed to identify and purify four apocarotenoids from fraction A4, representing the first report of such class of compound s in mass cultivated diatoms. In addition, fraction A5 resulted to be a mixture of oxylipins and fraction B4 of purine nucleosides, with adenosine as main component. Residual biomass was further processed to obtain an enriched lipidic extract, characterized by a high content of poly-unsaturated fatty acids. Anti-diabetic and anti-obesity activities were assessed by evaluating the expression levels of key proteins involved into thermogenesis (UCP1), lipolysis (Perilipin, HSL660), and mitochondrial dynamics (pDRP1, VDAC), on murine white adipocyte cell line. Overall, the data here reported confirmed S. pinnata as a platform for the discovery and purification of bioactive compounds with biomedical potential, and the biorefinery pipeline allow their combined isolation, thus advancing the sustainability of diatom exploitation for the discovery and production of bioactive molecules.

Neltuma nigra cotyledon and seed flour: nutritional, phytochemical, and techno-functional characterization and nutraceutic potential of polyphenolic enriched extract.

Neltuma nigra seed and cotyledon flours were prepared and characterized. Both flours showed high protein and fiber content, and low carbohydrate and fat content. The major storage proteins in N. nigra flours were albumin and globulin. In addition, flours contained carotenoids and potassium as the main micronutrients. The free and bound phenolic compounds, tannins and flavonoids were the major phytochemical components. The phenolic enriched extracts (PEE) of both flours contained apigenin-derived C-glycosyl flavones, including schaftoside, and isoschaftoside as the main components. The PEE from seed flour contained higher antioxidant capacity on ABTS˙+, FRAP, H2O2, O2˙- and higher inhibitory effect on α-amylase and α-glucosidase than the cotyledon flour extract. Techno-functional property analyses showed better water retention capacity and emulsifying stability for cotyledon flour and better oil holding capacity and emulsifying activity for seed flour. The results suggest better biological properties and a low-cost accessibility of seed flour, and a better macronutrient balance for cotyledon flour. Therefore, both can be considered as ingredients or functional food for special foods related to the metabolic syndrome and diseases related to oxidative processes.

Lycopene enriched tomato extract exerts modulatory effects on hypoxia, angiogenesis and metastasis during N-nitrosodiethylamine (NDEA) induced hepatocellular carcinoma in mice

Lycopene enriched tomato extract exerts modulatory effects on hypoxia, angiogenesis and metastasis during N-nitrosodiethylamine (NDEA) induced hepatocellular carcinoma in mice

A fast and simple procedure for the synthesis of a zinc and 1,4-benzene dicarboxylic acid metal-organic framework and its evaluation as a sorbent for dispersive micro solid phase extraction of pesticide residues.

In this work, Zn-1,4-benzenedicarboxylate metal-organic framework was synthesized by a simple hydrothermal process and used in dispersive micro solid phase extraction of some pesticide from various fruit juice and water samples. Seven widely consumed pesticides in agriculture (chlorpyrifos, haloxyfop-R-methyl, oxadiazon, diniconazole, clodinafop-propargyl, fenpropathrin, and fenaxoprop-p-ethyl) were selected as target analytes. In this work, dispersive micro solid phase extraction was followed by a liquid phase microextraction method to achieve more enrichment of the analytes, and the enriched analytes were quantified using a gas chromatography-flame ionization detector. The sorbent was authenticated by Fourier transform infrared spectrophotometry, X-ray diffraction, energy dispersive X-ray analysis, and scanning electron microscope imaging. The factors affecting the extraction efficiency of the developed method were investigated, and the validation of the method under the optimized extraction conditions presented satisfactory results for precision and trueness, with limits of detection and quantification in the ranges of 0.50-0.90 and 1.5-2.7 μg L-1, respectively. Enrichment factors and extraction recoveries were in the ranges of 239-392% and 47-78%, respectively. One river water and some fruit and vegetable juice samples were analyzed by the recommended method, and the obtained recoveries were between 90% and 102%.

Proteomic Analysis of Gingival Crevicular Fluid During Tooth Eruption

Objective: Gingival crevicular fluid (GCF) is a biological fluid that has the unique capacity to reflect changes in periodontium to its protein composition, making it ideal for potential biomarkers. There is limited information about the mechanism of tooth eruption, for which GCF might provide valuable knowledge. This study aimed to provide a proteomic approach to investigate the composition of GCF obtained from two different supraosseous tooth eruption stages of permanent molars changes.
 Methods: GCF samples were taken from a total of 26 healthy children, whose permanent molar just emerged from the gingiva (5-8 years old) and the occlusal equilibrium stage (9-13 years old). Proteins were extracted with Bio-Rad Rehydration Buffer followed by ZebaTM Spin Desalting Column. GCF samples were separated with two-dimensional polyacrylamide gel electrophoresis (2D-PAGE) followed by mass spectrometry- based protein identification.
 Results: A new optimized protocol with enriched protein extraction from GCF samples was developed. Several proteins related to tooth eruption were detected. It was determined that keratin type II cytoskeletal 4 and keratin type I cytoskeletal 9 decreased and albumin increased in GCF protein content in erupting teeth compared to those in the occlusal equilibrium phase.
 Conclusion: This methodology, which we have applied for 2D-PAGE of GCF, can also be a source for other studies. There is huge diagnostic potential in mass spectrometry technologies, this study can be carried forward by using other approaches.

Simultaneous Determination of Steroids and NSAIDs, Using DLLME-SFO Extraction and HPLC Analysis, in Milk and Eggs Collected from Rural Roma Communities in Transylvania, Romania.

This research aims to determine five steroids and four non-steroidal anti-inflammatory drugs in milk and egg samples collected from rural Roma communities in Transylvania, Romania. Target compounds were extracted from selected matrices by protein precipitation, followed by extract purification by dispersive liquid-liquid microextraction based on solidification of floating organic droplets. The extraction procedure was optimized using a 24 full factorial experimental design. Good enrichment factors (87.64-122.07 milk; 26.97-38.72 eggs), extraction recovery (74.49-103.76% milk; 75.64-108.60% eggs), and clean-up of the sample were obtained. The method detection limits were 0.74-1.77 µg/L for milk and 2.39-6.02 µg/kg for eggs, while the method quantification limits were 2.29-5.46 µg/L for milk and 7.38-18.65 µg/kg for eggs. The steroid concentration in milk samples was <MDL up to 4.30 µg/L, decreasing from 17α-ethinyl estradiol, 17β-estradiol, and estrone to estriol. The NSAID concentration was <MDL up to 3.41 µg/L, decreasing from ibuprofen, diclofenac, and ketoprofen to naproxen. The steroid concentration in the egg samples was <MDL to 2.79 µg/kg, with all steroids detected, while the concentration of NSAIDs was <MDL to 2.28 µg/kg, with only ibuprofen, ketoprofen, and naproxen detected. The developed protocol was successfully applied to the analysis of target compounds in real milk and egg samples.

Growth and Enzyme Application of Garlic Enriched With Zinc and Natural Magnesium

Objective: Garlic can help humans ensure healthy lives and promote well-being for all at all ages by sufficient zinc and magnesium intake.Method: Serpentine treated it by microwaving and sintering to enhance its crystallinity as well as its magnesium and zinc ion release rates. Furthermore, an enriched garlic enzyme extract had an approximately 8-fold increase in alliinase activity. Results: Strong bonding was observed for the microwaved and sintered powders, but also facilitated zinc ion reactions and reduced lattice defects. Accordingly, used for the garlic growth and enzyme experiments.Conclusions: (1) The sintered powder excellent magnesium and zinc ion release capability. (2) The enriched garlic enzymes had high alliinase activity, likely increasing the health benefits of the garlic.

A poly(2,6-diaminopyridine) functionalized magnetic porous carbon nanomaterial for extraction and determination of phenol, nitrophenols, and chlorophenols in water samples

Herein, for the first time, a new method for the simultaneous extraction of phenol (Ph), nitrophenols (NPs), and chlorophenols (CPs) as the priority pollutants was proposed using a magnetic porous carbon coated with poly(2,6-diaminopyridine). The analytes were separated/determined by an HPLC device coupled to a diode array detector. Magnetic porous carbon was obtained by the pyrolysis of MIL-53(Fe) as a precursor and then functionalized with a layer of poly(2,6-diaminopyridine). Next, the synthesized adsorbent was identified by various methods, including FT-IR, XRD, SEM, VSM, surface analysis, and TEM techniques. After that, the design of the experiment strategy was used to identify and optimize the extraction variables. The LODs and linear range were obtained in the range of 0.04–0.15 µg/L and 0.1–500 µg/L, respectively. The one-day RSDs and day-to-day RSDs were gained in the range of 4.2–9.5 % and 5.0–12.5 %, respectively. The enrichment factors and extraction recoveries were obtained in the range of 208–378 and 41.6–75.6 %, respectively. Eventually, the outlined method was employed to analyze diverse water/wastewater samples satisfactorily.

In-situ formation of CO2-incorportaed solid sorbent for dispersive solid phase extraction of phenolic compounds from water and wastewater samples prior to gas chromatography-flame ionization detector

BackgroundHerein, a new extraction procedure based on in-situ formation of carbon dioxide-incorporated solid sorbent was introduced for dispersive solid phase extraction of phenolic compounds from aqueous samples. In this study, incorporation of carbon dioxide into the structure of a diamine led to the formation of a solid compound in the sample solution that adsorbed the analytes. ResultsThe sample solution was mixed with isophorone diamine and placed under carbon dioxide stream. By doing so, isophorone diamine reacted with carbon dioxide and produced a carbamic acid analogue. It was dispersed into the sample solution as tiny particles that adsorbed the analytes. The adsorbed analytes were eluted by a volatile organic solvent and concentrated more by the vaporization of the eluate. The extraction procedure was done at low temperature to limit the releasing carbon dioxide from the produced compound. To obtain the reliable results, the method was validated and the obtained limits of detection and quantification were in the ranges of 0.29–41 and 0.96–1.3 ng/mL, respectively. Acceptable relative standard deviation (≤7.3%) and coefficient of determination (≥0.994) values confirmed the method repeatability and linearity. High enrichment factors (410–435) and extraction recoveries (82–87%) were attained with the introduced method. Significance and noveltyIn this work, a chemical reaction was done between isophorone diamine and carbon dioxide in solution. The produced product (sorbent) was insoluble in solution and dispersed in whole parts of the solution as tiny particles. A high contact area between the sorbent and analytes provided high extraction efficiency for the analytes. The method was successful utilized in determining target analytes in real samples and the matrix effect of the samples had no important effect on the obtained results.

Microextraction with smartphone detection of thiocyanate in saliva of tobacco smokers using paper-based analytical method.

This study presents a novel, cost-effective approach involving spectrophotometric and smartphone paper-based (SPB) methods and a distinctive salting-out air-assisted dispersive microextraction procedure to quantify thiocyanate in saliva samples. The method relies on the inhibitory effect of thiocyanate on quinoneimine dye formation during the Emerson reaction with sodium hypochlorite. Spectrophotometry quantifies the extracted dye by monitoring quinoneimine color intensity reduction at 525nm. In the SPB method, extracted dye is applied to a paper strip, a smartphone captures the colored paper, and an application analyzes red, green, and blue components. All analyte determination and extraction variables were explored. Both methods exhibit good linearity (10-100μg/L) with a coefficient of determination of 0.9991 and a limit of detection of 7.5μg/L for the spectrophotometric method, and a coefficient of determination of 0.9988 and a limit of detection of 8.8μg/L for the SPB method. The calculated values for the enrichment factor and extraction recovery of the developed extraction methodology were 46% and 93%, respectively. The methods detect thiocyanate in saliva samples, producing results comparable to a validated method.

In Situ Capture and Real-Time Enrichment of Marine Chemical Diversity.

Analyzing the chemical composition of seawater to understand its influence on ecosystem functions is a long-lasting challenge due to the inherent complexity and dynamic nature of marine environments. Describing the intricate chemistry of seawater requires optimal in situ sampling. Here is presented a novel underwater hand-held solid-phase extraction device, I-SMEL (In Situ Marine moleculELogger), which aims to concentrate diluted molecules from large volumes of seawater in a delimited zone targeting keystone benthic species. Marine benthic holobionts, such as sponges, can impact the chemical composition of their surroundings possibly through the production and release of their specialized metabolites, hence termed exometabolites (EMs). I-SMEL was deployed in a sponge-dominated Mediterranean ecosystem at a 15 m depth. Untargeted MS-based metabolomics was performed on enriched EM extracts and showed (1) the chemical diversity of enriched seawater metabolites and (2) reproducible recovery and enrichment of specialized sponge EMs such as aerothionin, demethylfurospongin-4, and longamide B methyl ester. These EMs constitute the chemical identity of each targeted species: Aplysina cavernicola, Spongia officinalis, and Agelas oroides, respectively. I-SMEL concentrated sponge EMs from 10 L of water in a 10 min sampling time. The present proof of concept with I-SMEL opens new research perspectives in marine chemical ecology and sets the stage for further sustainable efforts in natural product chemistry.

Ionic liquid-modified UiO-66-NH2 as sorbent of dispersive solid-phase extraction for rapid adsorption and enrichment of benzoylurea insecticides.

Ionic liquid (IL)-modified UiO-66-NH2 composite was prepared and used as sorbent of dispersed solid-phase extraction (dSPE) for extracting trace benzoylurea insecticides (BUs) from complex environmental matrices. The IL in framework endowed the prepared material had electropositive characteristics, which can produce interaction with electron rich guest molecules, such as BUs. The high thermal and chemical stability of UiO-66-NH2/IL enabled it to be reused for 16 times without significant reduction in adsorption performance. Due to the multiple forces including π-π, hydrogen bonding, and fluorine-fluorine interaction, UiO-66-NH2/IL showed good adsorption performance, short adsorption time (20 s) and rapid desorption ability (60 s) for BUs. Under the optimal conditions, the method exhibited wide linear range (0.02-500 ng mL-1) with correlation coefficient (R2) not worsethan 0.9928, high enrichment factor (252-300), and low detection limit (0.005-0.4 ng mL-1). The dispersed solid phase extraction coupling with high-performance liquid chromatography-diode array detector (dSPE-HPLC-DAD) was successfully used to detection of BUs in real environmental samples and satisfactory recoveries were obtained (80.5%±2.4-118%±3.2). The results indicated that UiO-66-NH2/IL composite can be a potential sorbent for the preconcentration of trace insecticidesin environmental samples.

Antiviral, virucidal and antioxidant properties of Artemisia annua against SARS-CoV-2

Natural products are a rich source of bioactive molecules that have potential pharmacotherapeutic applications. In this study, we focused on Artemisia annua (A. annua) and its enriched extracts which were biologically evaluated in vitro as virucidal, antiviral, and antioxidant agents, with a potential application against the COVID-19 infection. The crude extract showed virucidal, antiviral and antioxidant effects in concentrations that did not affect cell viability. Scopoletin, arteannuin B and artemisinic acid (single fractions isolated from A. annua) exerted a considerable virucidal and antiviral effect in vitro starting from a concentration of 50 µg/mL. Data from Surface Plasmon Resonance (SPR) showed that the inhibition of the viral infection was due to the interaction of these compounds with the 3CLpro and Spike proteins of SARS-CoV-2, suggesting that the main interaction of compounds may interfere with the viral pathways during the insertion and the replication process. The present study suggests that natural extract of A. annua and its components could have a key role as antioxidants and antiviral agents and support the fight against SARS-CoV-2 variants and other possible emerging Coronaviruses.

Combination of Ultrasound-Assisted Extraction and Ultrasound-Assisted Emulsification Microextraction for Separation and Enrichment of Permethrin and Deltamethrin Residues in Spinach Samples

Combination of Ultrasound-Assisted Extraction and Ultrasound-Assisted Emulsification Microextraction for Separation and Enrichment of Permethrin and Deltamethrin Residues in Spinach Samples

A Highly Selective Analytical Method Based on Salt-Assisted Liquid-Liquid Extraction for Trace-Level Enrichment of Multiclass Pesticide Residues in Cow Milk for Quantitative Liquid Chromatographic Analysis.

In this study, a simple, inexpensive, selective, and fast salting-out assisted liquid-liquid extraction (SALLE) technique coupled with high-pressure liquid chromatography-diode array detection (HPLC-DAD) was developed for the extraction, preconcentration, and analysis of trace level seven multiclass pesticide residues in pasteurized and raw cow milk samples. The significant factors that affect the extent to which the target analytes are extracted, such as the type of extraction solvent and its volume, the type and concentration of salting-out salts, the pH of the solution, and the extraction time, have been investigated. Under optimum conditions, the correlation coefficient (r2) was obtained within a range of 0.9982-0.9997 for a broad linear range concentration of 2-1500 ng·mL-1. Reliable sensitivity was achieved with limits of detection (LODs) and limits of quantification (LOQs) ranging from 0.58-2.56 ng·mL-1 and 1.95-8.51 ng·mL-1, respectively. While precision with interday and intraday in terms of relative standard deviations (RSDs) was observed in the range of 1.97 - 7.88% and 4.52 - 8.04%, respectively. The results of the precision studies reveal that good repeatability and reproducibility (RSDs <9) were achieved, thus showing a low variability extraction of the developed method. Finally, the proposed and validated approach was effectively used to extract and determine pesticide residues in real milk matrices; however, the target analytes were not detected in all samples.

Determination of releasing plasticizers from dental floss using ultrasound–assisted extraction–dispersive liquid–liquid microextraction followed by gas chromatography–flame ionization detector

Recently, the wide existenceof phthalate esters in healthcare products has attracted great attention from scientists due to their negative effects on human health. As a result, the current study was carried out to determine the presence of some plasticizers in dental floss such as di-n-butyl phthalate, diethyl phthalate, di-2-ethylhexyl phthalate, di-iso-butyl phthalate, and bis-2-ethylhexyl adipate. The studied analytes were extracted from dental floss by an ultrasound–assisted extraction procedure and then enriched by a dispersive liquid–liquid microextraction method. For this purpose, a piece of dental floss was immersed in acetonitrile under sonication to extract the analytes. Then, the acetonitrile phase containing the extracted analytes was mixed with a proper extraction solvent at µL–level and injected into sodium chloride solution. The extracted analytes were determined by gas chromatography–flame ionization detector and gas chromatography–mass spectrometry. Under the best extraction conditions, limits of detection and quantification were found in the ranges 0.20–0.33 and 0.67–1.1 µg/L in solution and 0.06–0.10 and 0.19–0.33 µg g−1 in dental floss, respectively. Relative standard deviations were in the ranges 4.0–6.5% and 5.1–7.9% for intra-day and inter-day precisions for five replicates at 60 µg/L of each analyte. Enrichment factors and extraction recoveries of the analytes ranged from 350 to 460 and 70–92%, respectively.

Application of temperature-sensitive hyperbranched polysiloxanes for the determination of polycyclic aromatic hydrocarbons in foodstuffs

In the present study, a novel temperature-sensitive hyperbranched polysiloxane dispersive liquid–liquid microextraction combined with high-performance liquid chromatography method (TS-HBPSi-DLLME-HPLC-UV) has been developed for the determination of five polycyclic aromatic hydrocarbons (PAHs) in instant noodles, in which temperature-sensitive hyperbranched polysiloxane (p(Ph-HBPSi-co-NIPAM)) was used as the extraction solvent. The method adsorbs the target analytes based on π-π and hydrophobic interaction between p(Ph-HBPSi-co-NIPAM) and PAHs, and then separates the polymer with the adsorbed target analytes from the solution by a phase transition. Microextraction parameters of the method including polymer ratio, type and concentration of salt, pH, extraction time and volume of acetonitrile were examined in detail and optimized. At the optimum conditions, the established method showed good linearity (R2 ≥ 0.9971) with satisfactory limits of detection (0.089–0.16 ng mL−1) and limits of quantitation (0.27–0.48 ng mL−1). The values for intra-day and inter-day precisions were lower than 3.2% and 4.4%, respectively. Enrichment factors (EFs) and extraction recoveries (ERs) ranged from 62 to 70 and 75 to 85%, respectively. Finally, the proposed microextraction technique was validated by analyte addition and recovery studies, and applied to the determination of five PAHs in instant noodles, demonstrating that temperature-sensitive hyperbranched polymers have great potential for application in sample pretreatment techniques.

An Interpretable High-Accuracy Method for Rice Disease Detection Based on Multisource Data and Transfer Learning.

With the evolution of modern agriculture and precision farming, the efficient and accurate detection of crop diseases has emerged as a pivotal research focus. In this study, an interpretative high-precision rice disease detection method, integrating multisource data and transfer learning, is introduced. This approach harnesses diverse data types, including imagery, climatic conditions, and soil attributes, facilitating enriched information extraction and enhanced detection accuracy. The incorporation of transfer learning bestows the model with robust generalization capabilities, enabling rapid adaptation to varying agricultural environments. Moreover, the interpretability of the model ensures transparency in its decision-making processes, garnering trust for real-world applications. Experimental outcomes demonstrate superior performance of the proposed method on multiple datasets when juxtaposed against advanced deep learning models and traditional machine learning techniques. Collectively, this research offers a novel perspective and toolkit for agricultural disease detection, laying a solid foundation for the future advancement of agriculture.

Hierarchical porous MOFs@COFs-based molecular trap solid-phase microextraction for efficient capture of PAHs and adsorption mechanisms

In view of the serious carcinogenic, teratogenic and mutagenic properties of polycyclic aromatic hydrocarbons (PAHs), which cause significant harm to human beings and the environment. Therefore, developing rapid adsorption and efficient analytical methods are research topic with practical significance. Herein, an innovative MOFs@COFs-based molecular trap materials were prepared with hierarchical porous core–shell structure and used as solid-phase microextraction (SPME) fiber coating, which combined with high performance liquid chromatography (HPLC) to achieve the efficient separation, capture, and detection for 7 PAHs in real samples. The possible adsorption mechanism was systematically verified and explained. Under the optimized conditions, the method developed in this experiment had a wide linear range (0.005–400 µg·L−1) with a good linear correlation coefficient (R2 > 0.9984), low detection limits (0.002–0.405 µg·L−1, S/N = 3) and low quantification limits (0.005–1.351 µg·L−1, S/N = 10). The relative standard deviations of individual fibers and different batches of fibers were 0.26–3.64% and 0.40–5.91%, respectively. Besides, the fiber showed higher enrichment capacity and extraction efficiency for PAHs compared to commercial SPME fibers and other material fibers, and exhibited excellent acid-base stability and reproducibility. This work provided a favorable synthetic pathway for the preparation of MOFs@COFs composites by layer-modified assembly synthesis strategy, showed excellent practical applicability as a SPME fiber coating combined with HPLC, and could be realized to provide technical support for future studies on the separation, adsorption, and analysis of PAHs in the petrochemical industrial zone of the Yellow River Basin.

Utilization of red and yellow Coffea arabica var. Caturra pulp: macronutrient analysis, carotenoid extraction, and encapsulation for dairy product enrichment.

This study aimed to investigate the macronutrient and carotenoid content of red and yellow Coffea arabica var. Caturra pulp, a by-product of coffee processing in Colombia. The study employed ultra-sound-assisted extraction (UAE) to extract carotenoids, and a 23 factorial design was used to evaluate the effects of pulp color, biomass-solvent ratio, and solvent mixture composition on carotenoid content and extraction yield. The condition that provided the highest carotenoid extraction was further encapsulated by spray drying and added to a dairy product. The results showed that coffee pulp has significant dietary fiber content and high levels of carotenoids, with yellow pulp having a higher content than red pulp. Lutein isomers and lutein esters were the most abundant carotenoids found in both red and yellow coffee pulp. The highest carotenoid extraction was achieved using a 1:40 (g/mL) biomass:solvent ratio and a 20:80% v/v Ethanol:Ethyl Acetate solvent mixture for the yellow pulp. The carotenoid extract also demonstrated high encapsulation efficiency (46.57 ± 4.03%) and was found to be stable when added to a fermented milk product. This study presents an alternative solution for utilizing coffee by-products in Colombia, which could positively impact the families of over half a million Colombian coffee producers.