A salt/salt aqueous two-phase system based on pH-switchable deep eutectic solvent for the extraction and separation of mulberry polysaccharides.

With the aim of expanding the potential application scope of mulberries, eleven pH-switchable deep eutectic solvents were screened for the ultrasonic-assisted extraction of mulberry polysaccharides, and a salt/salt aqueous two-phase system was constructed for the efficient separation of mulberry polysaccharides by regulating the system pH. DES-9 (tetraethylammonium chloride: octanoic acid molar ratio = 1: 2) with a critical response pH value of approximately 6.1 was concluded to be the best extraction solvent for extracting mulberry polysaccharides. A maximum polysaccharide extraction yield of 270.71 mg/g was obtained under the optimal conditions. The maximum polysaccharide extraction efficiency was 78.09 % for the pH-driven tetraethylammonium chloride/K2HPO4 aqueous two-phase system. An acidic ß-pyran mulberry polysaccharide with a low-molecular weight of 9.26 kDa and a confirmed monosaccharide composition were obtained. This efficient and environmentally friendly polysaccharide separation method offers a new approach for the efficient extraction and utilization of other plant polysaccharides.

Emerging deep eutectic solvents for food waste valorization to achieve sustainable development goals: Bioactive extractions and food applications.

Food waste, accounting for about one-third of the total global food resources wasted each year, is a substantial challenge to global sustainability, contributing to adverse environmental impacts. The utilization of food waste as a valuable source for bioactive extraction can be facilitated through the application of DES (Deep Eutectic Solvents). Acknowledging the significant need to tackle this issue, the United Nations integrated food waste management into its Sustainable Development Goals, hence, the present review explores the role of DES in bioactive compounds extraction from food waste. Various extraction processes using the DES system are thoroughly studied and the application of bioactive components as antioxidants, antimicrobials, flavourings, nutraceuticals, functional ingredients, additives, and preservatives is investigated. Most importantly, regulatory considerations and safety aspects of DES in food applications are discussed in-depth along with consumer perception and acceptance of DES in the food sector. The key hypothesis of the review is to evaluate emerging DES systems for their efficiency in bioactive extraction technologies and various food applications. Overall, this review provides a comprehensive understanding of utilizing DES for synthesizing valuable food waste-derived bioactive components, offering a sustainable approach to waste management and the development of high-value products.

Supramolecular deep eutectic solvents: Current advances and critical evaluation of cyclodextrins from solute to solvent in emerging functional food.

The acceptance of nonconventional solvents as viable substitutes for traditional organic solvents has been widely recognized in order to comply with food-safety and sustainability regulations. Cyclodextrins (CDs), derived from starch, are cyclic oligosaccharides with the ability to form inclusion complexes with a variety of functional substances as the result of their distinctive structure, which enables them to effectively encapsulate bioactive compounds, rendering them highly sought after for use in food applications. In the implementing plan to achieve carbon-neutral emissions by 2050, the novel generation of supramolecular deep eutectic solvents (SUPRADES) has garnered increased attention and interest. The approach of utilizing SUPRADES as emerging solvents was just beginning to be applied to food studies. This review summarizes a revision of the current advances and critical evaluation of cyclodextrin-based SUPRADES (CD-based SUPRADES) as promising solvents for the enhancement of the extraction efficiency, solubilization and stability of bioactive compounds, adsorption and separation of food components, packaging materials, and modification of biopolymers. To meet the sustainable processing needs of the food industry, the emerging categories of CD-based SUPRADES need to be further fabricated. Herein, our review will sort out the potential application of CD-based SUPRADES in the food industry, aiming to provide a better understanding of CD-based SUPRADES within the viewpoint of food science. Formulation intricacies and scalability issues in real functional foods using CD-based SUPRADES as media need more efforts.

Development of Thin Film Microextraction with Natural Deep Eutectic Solvents as 'Eutectosorbents' for Preconcentration of Popular Sweeteners and Preservatives from Functional Beverages and Flavoured Waters.

An eco-friendly method for the determination of sweeteners (aspartame, acesulfame-K) and preservatives (benzoic acid, sorbic acid, methylparaben, ethylparaben) in functional beverages and flavoured waters using thin film microextraction (TFME) and high-performance liquid chromatography with UV detection (HPLC-UV) was proposed. A series of fourteen green and renewable solidified natural deep eutectic solvents (NADESs) were prepared and tested as 'eutectosorbents' in TFME for the first time. In the proposed method, the NADES containing acetylcholine chloride and 1-docosanol at a 1:3 molar ratio was finally chosen to coat a support. Four factors, i.e., the mass of the NADES, pH of the samples, extraction time, and desorption time, were tested in the central composite design to select the optimal TFME conditions. Limits of detection were equal to 0.022 µg mL-1 for aspartame, 0.020 µg mL-1 for acesulfame-K, 0.018 µg mL-1 for benzoic acid, 0.026 µg mL-1 for sorbic acid, 0.013 µg mL-1 for methylparaben, and 0.011 µg mL-1 for ethylparaben. Satisfactory extraction recoveries between 82% and 96% were achieved with RSDs lower than 6.1% (intra-day) and 7.4% (inter-day). The proposed 'eutectosorbent' presented good stability that enabled effective extractions for 16 cycles with recovery of at least 77%. The proposed NADES-TFME/HPLC-UV method is highly sensitive and selective. However, the use of a solid NADES as a sorbent, synthesized without by-products, without the need for purification, and with good stability on a support with the possibility of reusability increases the ecological benefit of this method. The greenness aspect of the method was evaluated using the Complex modified Green Analytical Procedure Index protocol and is equal to 84/100.

Advances in Extracting Bioactive Compounds from Food and Agricultural Waste and By-Products Using Natural Deep Eutectic Solvents: A Circular Economy Perspective.

Due to the urgent need for a transition to sustainable, zero-waste green technology, the extraction of bioactives from food and agricultural by-products and waste has garnered increasing interest. Traditional extraction techniques often involve using organic solvents, which are associated with environmental and health risks. Natural deep eutectic solvents (NADESs) have emerged as a promising green alternative, offering advantages such as low toxicity, biodegradability, and the ability to dissolve a wide range of biomolecules. This review provides a comprehensive overview of recent trends in the application of NADESs for extracting bioactive compounds from sustainable sources. The review explains the composition and principles of preparation and highlights various applications of NADESs in extracting different classes of bioactive compounds, emphasizing their potential to revolutionize extraction processes. By summarizing the latest advancements and trends, this review aims to support research and industrial applications of NADESs, promoting more sustainable and efficient extraction methods in the food and agricultural sectors.

A Comprehensive Review on Deep Eutectic Solvents: Their Current Status and Potential for Extracting Active Compounds from Adaptogenic Plants.

Deep eutectic solvents (DESs) have attracted attention from researchers as novel compounds for extracting active substances because of their negligible toxicity, polarity, and ability to be tailored depending on the experiment. In this review, we discuss deep eutectic solvents as a promising medium for the extraction of adaptogenic compounds. In comparison to traditional methods, extraction with the use of DESs is a great alternative to the excessive usage of harmful organic solvents. It can be conducted in mild conditions, and DESs can be designed with different precursors, enhancing their versatility. Adaptogenic herbs have a long medicinal history, especially in Eastern Asia. They exhibit unique properties through the active compounds in their structures, including saponins, flavonoids, polysaccharides, and alkaloids. Therefore, they demonstrate a wide range of pharmaceutical effects, such as anti-inflammatory, antibacterial, and anticancer abilities. Since ancient times, many different adaptogenic herbs have been discovered and are well known, including Panax ginseng, Scutellaria baicalensis, and Schisandra chinensis. Active compounds can be extracted using standard methods, such as hydrolyzation, maceration, and conventional reflux extraction. However, due to the limitations of classical processing technologies, there has been a need to develop new and eco-friendly methods. We focus on the types of solvents, extraction efficiency, properties, and applications of the obtained active compounds. This review highlights the potential of DESs as eco-friendly alternatives for extracting bioactive compounds.

Preconcentration of Cinnamic Acid Derivatives in Traditional Chinese Medicines by an Effective Dispersive Liquid-Liquid Microextraction Based on a Hydrophobic Deep Eutectic Solvent.

A dispersive liquid-liquid microextraction based on hydrophobic deep eutectic solvent (hDES) was developed for the extraction and quantification of four cinnamic acid derivatives in traditional Chinese medicines coupled with high-performance liquid chromatography-ultraviolet detection. In this method, a hDES (tetrabutylammonium chloride-hexanoic acid, molar ratio of 1:2) was prepared as the extractant. It only took 15 s to handle multiple samples simultaneously by hand-assisted dispersion. The use of a narrow-bore tube reduced the amount of the hydrophobic extractant with easier recovery. The approach was influenced by several key parameters, including the composition and consumption of the DES, sample phase pH, salt amount, extraction time, and centrifugation time, all of which had been investigated and optimized. Moreover, the formation of the DES was characterized by Fourier-transform infrared spectroscopy and differential scanning calorimetry. Under the optimal conditions, enrichment factors of the target analytes ranged from 135 to 220. Satisfactory linearities (r ≥ 0.9977), detection limits (0.2-0.4 ng/mL), precision (<8.5%), and accuracy (recoveries: 90.0%-104.6%) were obtained. The method has been successfully applied to the simultaneous extraction and preconcentration of four cinnamic acid derivatives in Chinese medicinal samples with rapidness, high efficiency, and convenience.

Exploration of the Solubility Hyperspace of Selected Active Pharmaceutical Ingredients in Choline- and Betaine-Based Deep Eutectic Solvents: Machine Learning Modeling and Experimental Validation.

Deep eutectic solvents (DESs) are popular green media used for various industrial, pharmaceutical, and biomedical applications. However, the possible compositions of eutectic systems are so numerous that it is impossible to study all of them experimentally. To remedy this limitation, the solubility landscape of selected active pharmaceutical ingredients (APIs) in choline chloride- and betaine-based deep eutectic solvents was explored using theoretical models based on machine learning. The available solubility data for the selected APIs, comprising a total of 8014 data points, were collected for the available neat solvents, binary solvent mixtures, and DESs. This set was augmented with new measurements for the popular sulfa drugs in dry DESs. The descriptors used in the machine learning protocol were obtained from the σ-profiles of the considered molecules computed within the COSMO-RS framework. A combination of six sets of descriptors and 36 regressors were tested. Taking into account both accuracy and generalization, it was concluded that the best regressor is nuSVR regressor-based predictive models trained using the relative intermolecular interactions and a twelve-step averaged simplification of the relative σ-profiles.

Lipase Substrate Anchoring Dynamics Guided the Rational Design of Novel Deep Eutectic Solvents for Glucose Ester Synthesis.

Glucose esters, heralded for their emulsifying and solubilizing properties, have found increasing application in food, pharmaceutical, and cosmetic sectors. However, the environmental impact of chemical synthesis and the problem of enzyme inactivation in enzymatic synthesis have hindered their application. To this end, deep eutectic solvents (DESs) with stabilized enzyme activity and better solubility properties are considered a promising solution. In the study, 12 kinds of hydrophilic DESs and 22 kinds of hydrophobic DESs were screened, and we examined their efficiency in the synthesis of glucose esters. The results indicated that d,l-menthol-based DESs have excellent performance, with the highest yield of 92.85%. The molecular dynamics simulations suggested that it can be attributed to DESs changing the size of substrate binding pocket, which in turn affects the substrate anchoring ability and the catalytic efficiency of lipase. To further evaluate the effects of the solvent on the substrate anchoring ability of enzymes, a novel strategy, namely, substrate anchoring index, which is based on the stability of the tetrahedral intermediate, was proposed and used for the designing of more efficient DESs. Fortunately, novel DESs based on cyclohexanone (menthol analogues) were successfully developed with a yield of 98.85%.

New Green Biorefinery Strategies to Valorize Bioactive Fractions from Palmaria palmata.

A biorefinery process was developed to isolate phycobiliproteins, sulfated polysaccharides, and phenolic compounds from Palmaria palmata. The extraction process was carried out in three stages using ultrasound-assisted extraction (UAE) and pressurized liquid extraction (PLE) integrated with different natural deep eutectic solvents (NaDESs). In general, PLE provided higher phycobiliprotein contents than UAE in the first step of the process. In fact, the hydrolysis product of the PLE-NaDES extracts achieved a higher antioxidant capacity than that of the UAE-NaDES extracts. Particularly, glycerol:glucose (2:1) with 50% water in combination with PLE was the most suitable NaDES to recover the highest phycobiliprotein, protein, and sulfated polysaccharide contents from Palmaria palmata in the first and second steps of the biorefinery process. Finally, a PLE-NaDES using choline chloride:glycerol (1:2) with 60% water as the NaDES was employed for the recovery of antioxidant and neuroprotective phenolic compounds from the residue of the second step, obtaining a higher total phenolic content than employing PLE with ethanol/water (70:30, v/v) as the extraction solvent. Moreover, a forced stability study revealed that the NaDESs provided a protective effect compared to the water extracts against the degradation of phycobiliproteins, preserving their color over time. This study contributes to the recovery of high-value components from an undervalued biomarine source through a sustainable biorefinery process.

Novel stationary basket reactor for effective biomass delignification with deep eutectic solvent.

Biomass pretreatment and conversion are crucial for sustainable development, but lack information on equipment that ensures effective mass transfer and easy biomass separation post-process. This work introduces a novel basket reactor with a stationary bed (StatBioChem) for biomass processing using deep eutectic solvents (DESs). We compared the delignification efficiencies of soft and hard biomass samples processed in the StatBioChem reactor, a stirred tank reactor (STR), and a commercial SpinChem® reactor. The StatBioChem design allowed DES to flow evenly through biomass in the basket, achieving the highest delignification degree, particularly for hard biomass. This effect was not observed in the SpinChem® basket reactor. High delignification led to increased glucose yields in subsequent enzymatic hydrolysis. The StatBioChem effectively combines the simplicity and efficiency of an STR with the ease of solvent recovery typical of basket reactors.

Enhanced furfural extraction using neoteric hydrophobic solvents for sustainable biomass recovery and bioenergy applications.

The recovery of furfural from hemicellulosic biowastes is important for developing sustainable and renewable energy alternatives to fossil fuels. However, current methods are inefficient and environmentally questionable. To address this issue, this study introduces neoteric hydrophobic solvents, specifically deep eutectic solvents (DESs) and ionic liquids (ILs). Of the 32 solvents tested, thymol:decanoic acid 1:1 (Thy:DecA) DES and trihexyltetradecyl phosphonium bis(trifluoro methylsulfonyl) imide [P14,6,6,6][NTf2] IL were the most effective, with extraction efficiencies of 94.1% and 97.1%, respectively. These solvents outperformed the reference solvent toluene, with an efficiency of 81.2%, while also showing favorable characteristics in multiple investigated criterions. For the first time, excellent performance stability was demonstrated under various operational conditions and reusability over multiple extraction and regeneration cycles. Furthermore, to provide insights into the molecular mechanisms of extraction, computational quantum chemistry modeling was employed, which showed a strong agreement with the experimental results. The development of these new neoteric solvents for furfural recovery from biowaste offers a highly effective, sustainable, and eco-friendly alternative to traditional solvents, representing a significant breakthrough in the field of renewable bioenergy production and sustainable materials recovery.

Sustainable reuse of date palm biomass via extraction of cellulose using natural deep eutectic solvent (NaDES) and microwave-assisted process.

In Saudi Arabia, date palm biomass is often considered bulk waste, posing health risks to humans and the environment when incinerated or disposed of in landfills. This study explores the reuse of date palm biomass through cellulose extraction using Natural Deep Eutectic Solvent (NaDES) and Microwave-assisted process as compared with conventional alkali peroxide method. Characterization of the extracted cellulose was conducted using scanning electron microscopy (SEM), thermogravimetric analysis (TGA), differential scanning calorimetry (DSC), x-ray diffractometry (XRD), and fourier-transform infrared spectroscopy (FTIR). Our NaDES-microwave method produced a cellulosic product with a yield of 89 %, double that of conventional non-microwave methods and more rapid cellulosic extraction, indicating its favorable efficiency. Moreover, the incorporation of the microwave process resulted in larger cellulosic crystallite sizes and higher thermal stabilities in the produced samples. The utilization of NaDES and microwave-assisted process for cellulose extraction from date palm biomass not only demonstrates superior efficiency, but also accentuates a significant sustainability advantage by repurposing a prevalent waste stream, thereby mitigating environmental and health risks associated with traditional disposal methods.

Natural deep eutectic solvent-functionalized mesoporous graphitic carbon nitride-reinforced electrospun nanofiber: a promising sorbent in miniaturized on-chip thin film micro-solid-phase extraction prior to liquid chromatography-tandem mass spectrometry for measuring NSAIDs in saliva.

To meet the needs of developing efficient extractive materials alongside the evolution of miniaturized sorbent-based sample preparation techniques, a mesoporous structure of g-C3N4 doped with sulfur as a heteroatom was achieved utilizing a bubble template approach while avoiding the severe conditions of other methods. In an effort to increase the number of adsorption sites, the resultant exfoliated structure was then modified with thymol-coumarin NADES as a natural sorbent modifier, followed by introduction into a nylon 6 polymer via an electrospinning process. X-ray diffraction (XRD), Fourier transform infrared (FT-IR) spectroscopy, field-emission scanning electron microscopy (SEM), energy-dispersive X-ray spectroscopy (EDS), and Brunauer-Emmett-Teller (BET) surface area analysis validated S-doped g-C3N4 and composite production. The prepared electrospun fiber nanocomposite, entailing satisfactory processability, was then successfully utilized as a sorbent in on-chip thin film micro-solid-phase extraction of non-steroidal anti-inflammatory drugs (NSAIDs) from saliva samples prior to liquid chromatography-tandem mass spectrometry (LC-MS/MS) analysis. Utilizing a chip device, a thin film µ-SPE coupled with LC-MS/MS analysis yielded promising outcomes with reduced sample solution and organic solvents while extending lifetime of a thin film sorbent. The DES-modified S-doped g-C3N4 amount in electrospun was optimized, along with adsorption and desorption variables. Under optimal conditions, selected NSAIDs were found to have a linear range of 0.05-100.0 ng mL-1 with an R2 ≥ 0.997. The detection limits were ranged between 0.02 and 0.2 ng mL-1. The intra-day and inter-day precisions obtained were less than 6.0%. Relative recoveries were between 93.3 and 111.4%.

Spectrophotometric determination for green hydrophobic deep eutectic solvent-based microextraction of Brilliant Blue FCF (E133) from beverages.

In this study, a simple, sensitive, and rapid method called green hydrophobic deep eutectic solvent-based liquid-liquid microextraction was developed to extract Brilliant Blue FCF dye from beverages. This method utilizes hydrophobic DES obtained by forming tetrabutylammonium bromide and 1-octanol in a 1:5 ratio as green extraction solvent. The transition of Brilliant Blue FCF to the DES phase occurred on its own, without the need for any reagents such as added salt or tetrahydrofuran. Several crucial factors were tried to get the best extraction efficiency, including species, DES volume and molar ratio, solution pH, ultrasonication, and centrifugation time. Under optimum conditions, extraction recoveries were achieved in the range of 95.1-101.3 % with the method developed for Brilliant Blue FCF. The detection and determination limits were observed to be 4.1 µg l-1 and 12.1 µg l-1, respectively. In addition, the relative standard deviation values for the method's accuracy were found to be 2.23 % and 3.48 % within and between days, respectively. It has been established that the developed method is highly environmentally friendly thanks to the application of the Analytical GREENness (AGREE) and Green Analytical Procedure Index (GAPI) tools. This study shows that DES applications can be carried out without the use of emulsifiers and dispersants by prioritizing the use of hydrophobic DES compounds as environmentally friendly and green extraction solvents in food samples.

Preparation of straw/PLA composites with excellent flame retardancy based on deep eutectic solvent sulfonation.

There have been numerous studies on flame retardant modification of natural fiber/PLA composite materials due to the demand for applications. However, the existing flame retardant modification methods mostly involve adding flame retardants, which have a negative impact on the mechanical properties. Based on this, this study aims to introduce sulfonic groups into the cellulose of straw fibers via modification with a sulfamic acid-based deep eutectic solvent (SDES), thereby achieving flame retardance without affecting the inherent mechanical properties of the composite material. The performance enhancement of DS/PLA is manifested in the following specific aspects: the LOI reaches 36.53 %, the thermal stability is improved from 7.8 % of the residual carbon of PS/PLA to 38.4 %, and the tensile modulus is increased by 69.5 %. The preparation scheme for straw/PLA composite materials in this study is simple, economical, and efficient, and the flame retardant performance of the composite material is excellent, providing valuable references for flame retardant modification of natural fiber/plastic composite materials.

High-performance removal of methylene blue dye using porous lignin extracted from sugarcane bagasse by deep eutectic solvent.

This study evaluated the ability of triethyl benzyl ammonium chloride/lactic acid deep eutectic solvent extracted lignin (TEBAC/LA-DES-L) to adsorb methylene blue (MB) without additional functional group modification. The structure and morphology of TEBAC/LA-DES-L were characterized using SEM, BET, FT-IR, and TGA techniques. Various factors influencing MB adsorption, such as extraction temperature, solution pH, adsorbent dose, initial MB concentration, adsorption time, and reaction temperature, were investigated. The Redlich-Peterson isotherm displayed a good fit for the experimental data, with a maximum adsorption capacity of 85.16 mg/g. Kinetic analysis suggested that the adsorption process followed the pseudo-second-order model, with adsorption occurring in <100 min on DES-L-4 h. The mechanism of MB adsorption on DES-L-4 h was attributed to electrostatic attraction, hydrophobic interactions, and hydrogen bonding forces. Overall, DES-L-4 h demonstrated high adsorption capacity and rapid adsorption rate, making it a promising adsorbent for effectively removing cationic dyes from wastewater.

Multifunctional composite film of curcumin Pickering emulsion stabilized by lignocellulose nanofibrils isolated from bamboo shoot shells for monitoring shrimp freshness.

Concerns about food safety and environmental impact from chemical surfactants have prompted interest in natural lignocellulosic materials as alternatives. In this study, we combined hydrated deep eutectic solvent (DES) pretreatment with ultrasound treatment to prepare lignocellulosic nanofibrils (LCNF) from bamboo shoot shells with appropriate surface properties for stabilizing Pickering emulsions. The pretreatment intensity effectively modulated the surface characteristics of LCNF, achieving desirable wettability through lignin retention and in-situ esterification. The resulting LCNF/curcumin Pickering emulsion (CPE) demonstrated curcumin protection and pH-responsive color changes, while the ensuing CPE/PVA composite film exhibited ultraviolet shielding, mechanical strength, oxygen barrier, and antioxidant properties. Furthermore, the CPE/PVA film showed promise as a real-time indicator for monitoring shrimp freshness, maintaining sensitivity to spoilage even after six months of storage. These findings advance the advancement of green LCNF technologies, providing eco-friendly solutions for valorizing bamboo shoot shells and enhancing the application of LCNF in Pickering emulsions.

Polyol-assisted ternary deep eutectic solvent protective lignocellulose pretreatment for high-efficiency xylan utilization and ethanol production.

The traditional lignocellulose pretreatment by deep eutectic solvent (DES) was usually conducted under higher acidic, alkaline and high temperature conditions, which leads to the severe degradation of xylan, decreasing the subsequent reducing sugar concentration by enzymatic hydrolysis and further ethanol fermentation. It is essential to develop an effective DES that selectively removes lignin while preventing excessive xylan degradation during lignocellulose pretreatment. An effective ethylene glycol-assisted ternary DES was designed to treat corn straw (CS) at 100 °C for 6 h. 65.51 % lignin removal was achieved, over 93.46 % cellulose and 50.22 % xylan were retained in pretreated CS with excellent enzymatic digestibility (glucan conversion of 77.05 % and xylan conversion of 71.72 %), total sugar conversion could reach 75.93 %, implying the unique capacity to selectively remove lignin while preserving carbohydrate components. Furthermore, the universality of the selective removal of lignin and effective retention of xylan by ternary DES has been successfully proven by other polyols. The enzymatic hydrolysate of ternary DES-pretreated CS fermented over our genetically engineered yeast strain SFA1OE gave a high ethanol yield of 0.488 g/g total reducing sugar, demonstrating the effectiveness of the polyol-assisted ternary DES pretreatment in achieving high-efficiency cellulosic ethanol production.

Optimization and Validation of a Cheaper, Safer, and More Sustainable Methodology for Aflatoxins Determination in Rich-Lipidic Matrices (Pistachio Nuts) Using Deep Eutectic Solvent Extraction and UHPLC-FLD Analysis.

Aflatoxins pose a major health concern and require strict monitoring in food products. Existing methods rely on hazardous organic solvents for extraction, prompting the development of a greener alternative. This study explores deep eutectic solvents (DESs) for aflatoxin extraction from pistachios, a valuable food product prone to aflatoxin contamination. The proposed method utilizes DES extraction followed by solid-phase extraction cleanup and ultrahigh-performance liquid chromatography coupled with fluorescence detector analysis. Recovery rates ranged from 85.5 to 99.1% for pistachios spiked with 1-8 ng/g aflatoxins, in compliance with EU regulations, with coefficients of variation less than 2.94%. The method demonstrates good sensitivity with limits of detection and quantification in the range of 0.02-0.22 ng/g and 0.05-0.72 ng/g, respectively. Greenness assessment using AGREEPrep and White Analytical Chemistry metrics confirms its environmental sustainability. This approach offers a promising, safer, and more eco-friendly alternative for aflatoxin extraction from complex food matrices like pistachios.