New robust and efficient liquid membranes for conductive vial electromembrane extraction of acids with low to moderate hydrophilicity in human plasma.

The current paper reports two new, robust, and efficient conditions for electromembrane extraction of acidic substances from human plasma. Two systems were developed based on eutectic solvents: A1 ("A" for acid) comprised dodecyl methyl sulfoxide and thymol in 1:2 ratio (w/w) as liquid membrane, while A2 used [6-methylcoumarin:thymol (1:2)]:2-nitrophenyl octyl ether in 2:1 ratio (w/w). The performance of A1 and A2 was characterized by extraction of 31 acidic model analytes (pharmaceutical drugs and nutrients) spiked into 100 µL human plasma diluted 1:1 (v/v) with phosphate buffer pH 7.4. The acceptor solution was 50 mM NH4HCO3 buffer pH 10.0, and extraction was performed at an agitation rate of 750 RPM. Voltage and extraction time were 30 V for 30 min and 10 V for 20 min for A1 and A2, respectively. Under optimal conditions, A1 extracted analytes with 1.8 ≤ log P ≤ 6.0 with an average recovery (R) of 85.1%, while A2 extracted in a range of 0.5 ≤ log P ≤ 6.0 with an average recovery of 79.9%. Meanwhile, extraction current was low at 9 and 26 µA, respectively, which is indicative of good system robustness. Using UHPLC-MS/MS analysis of the acceptor solution, repeatability of the A1 and A2 methods was determined to be 2.8-7.7% and 3.3-9.4% for R > 40%, matrix effects were 82-117% and 84-112%, respectively, and linear calibration curves were obtained. The performance and compatibility with human plasma represent a major improvement over previous state-of-the-art liquid membranes for acidic analytes, namely 1-octanol.

Electromembrane extraction of peptides based on charge, hydrophobicity, and size - A large-scale fundamental study of the extraction window.

This study investigated the capability of electromembrane extraction (EME) as a general technique for peptides, by extracting complex pools of peptides comprising in total of 5953 different substances, varying in size from seven to 16 amino acids. Electromembrane extraction was conducted from a sample adjusted to pH 3.0 and utilized a liquid membrane consisting of 2-nitrophenyl octyl ether and carvacrol (1:1 w/w), containing 2% (w/w) di(2-ethylhexyl) phosphate. The acceptor phase was 50 mM phosphoric acid (pH 1.8), the extraction time was 45 min, and 10 V was used. High extraction efficiency, defined as a higher peptide signal in the acceptor than the sample after extraction, was achieved for 3706 different peptides. Extraction efficiencies were predominantly influenced by the hydrophobicity of the peptides and their net charge in the sample. Hydrophobic peptides were extracted with a net charge of +1, while hydrophilic peptides were extracted when the net charge was +2 or higher. A computational model based on machine learning was developed to predict the extractability of peptides based on peptide descriptors, including the grand average of hydropathy index and net charge at pH 3.0 (sample pH). This research shows that EME has general applicability for peptides and represents the first steps toward in silico prediction of extraction efficiency.

Parallel electromembrane extraction of peptides with monoterpene and medium-length fatty acid deep eutectic solvents.

BACKGROUND: Electromembrane extraction (EME) involves the process of mass transfer of charged analytes from an aqueous sample through an organic liquid membrane into an aqueous acceptor medium under the influence of an electrical field. Successful solvation of the analyte within the liquid membrane is of paramount importance and involves molecular interactions with the liquid membrane. In this comprehensive investigation, parallel EME was examined using a training set of 13 model peptides employing deep eutectic solvents as the liquid membrane. These deep eutectic solvents were formulated by mixing specific monoterpenes (thymol, menthol, camphor) with medium-chain fatty acids (1-octanoic acid and 1-decanoic acid). RESULTS: From an array of different liquid membrane compositions explored, it was revealed that the combination of camphor and 1-decanoic acid (in a 1:1 w/w ratio) with 2% di (2-ethylhexyl) phosphate (DEHP) delivered the most efficient extraction system. The solvation of the model peptides within this liquid membrane predominantly relied on ionic interactions between protonated basic functionalities and DEHP, along with hydrogen bond interactions between the deprotonated acid functionalities (hydrogen bond acceptor) and 1-decanoic acid (hydrogen bond donor). Selectivity was modulated by the pH of the sample and acceptor solutions, with a direct correlation to the polarity and net charge of the model peptides. The ionization of 1-decanoic acid in the interfacial region between the sample and liquid membrane emerged as an important factor influencing the selectivity. SIGNIFICANCE AND NOVELTY: Although parallel EME of peptides has been reported previously, the current liquid membrane provides an extraction system with sufficient stability for the first time. Selective extraction of peptides through EME holds substantial promise within the realm of next-generation environmentally-friendly sample preparation methodologies. The findings presented in this paper contribute significantly to our fundamental understanding of these processes, and may serve as an important reference for the development of future methods in this field.

Electromembrane extraction of peptides based on hydrogen bond interactions.

BACKGROUND: Electromembrane extraction (EME) of peptides reported in the scientific literature involve transfer of net positively charged peptides from an aqueous sample, through a liquid membrane, and into an aqueous acceptor solution, under the influence of an electrical field. The liquid membrane comprises an organic solvent, containing an ionic carrier. The purpose of the ionic carrier is to facilitate peptide solvation in the organic solvent based on ionic interactions. Unfortunately, ionic carriers increase the conductivity of the liquid membrane; the current in the system increases, the electrolysis in sample and acceptor is accelerated, and the extraction system tend to be unstable and suffers from drifting pH. RESULTS: In the present work, a broad selection of organic solvents were tested as pure liquid membrane for EME of peptides, without ionic carrier. Several phosphates provided high mass transfer, and tri(pentyl) phosphate was selected since this solvent also provided high operational stability. Among 16 different peptides used as model analytes, tri(pentyl) phosphate extracted those with net charge +1 and with no more than two polar side chains. Tri(pentyl) phosphate served as a very strong hydrogen bond acceptor, while the protonated peptides were hydrogen bond donors. By such, hydrogen bonding served as the primary interactions responsible for mass transfer. Tri(pentyl) phosphate as liquid membrane, could exhaustively extract leu-enkephalin, met-enkephalin, and endomorphin from human blood plasma and detected by LC-MS/MS. Calibration curves were linear (r2 > 0.99) within a concentration range from 1 to 500 ng/mL, and a relative standard deviation within 12% was observed for precision studies. SIGNIFICANCE: The current experiments are important because they indicate that small peptides of low polarity may be extracted selectively in EME based on hydrogen bond interactions, in systems not suffering from electrolysis.

Generic Liquid Membranes for Electromembrane Extraction of Bases with Low or Moderate Hydrophilicity.

For the first time, this paper introduces the idea of generic extraction conditions in electromembrane extraction (EME), where the selection of the liquid membrane is based on the charge (z) and hydrophobicity (log P) of the analyte. A broad range of organic solvents were tested as liquid membranes, and 90 basic pharmaceuticals were used as model analytes (-4.2 < log P < 8.1). 2-Nitrophenyl octyl ether (NPOE) was confirmed as a highly efficient liquid membrane for mono- and dibases (+1.0 ≤ z ≤ +2.0) of low polarity in the log P range of 2.2-6.4. This log P range was set as the extraction window (operational range) of NPOE. NPOE provided very high operational stability. At 50 V, the current was at a 1 µA level, and gas formation and drifting pH due to electrolysis were insignificant. 2-Undecanone was discovered as a new and robust alternative. This solvent extracted monobasic analytes (z = +1) in the log P range of 1.0-5.8 and was efficient even for bases of moderate polarity. The current was at the 1-3 µA level when 2-undecanone was operated at 50 V. Tri(pentyl) phosphate emerged as another new alternative for bases in the log P range of 0.5 to 5.5, providing greater selectivity differences. This solvent provided a higher current (30-50 µA), but the EME system stability was not compromised. 2-Undecanone and tri(pentyl) phosphate extracted protonated bases mainly by hydrogen bond interactions. NPOE, on the other hand, extracted based on a combination of hydrogen bond and π-type interactions and was consequently less selective.

A green microfluidic method based liquid phase microextraction for the determination of parabens in human urine samples.

Development of green approaches have emerged as a challenge that highlight the pressing need for nontoxic solvents, miniaturized method and bio-degradable materials. In this regard, an environmentally-friendly microfluidic system based on natural deep eutectic solvents (DESs) immobilized in agarose membranes was developed to extract parabens from urine samples for the first time. A comprehensive study of the support liquid membrane showed that only 3 µL of camphor and thymol (2:1 molar ratio) was an interesting option as a substitute for conventional (toxic) solvents used to date. Other experimental conditions were optimized and pH 4 (HCl) and 12 (NaOH) were selected as sample and acceptor solution, respectively. Both solutions (sample and acceptor) were fixed at 1 µL min-1 as flow rate. The proposed green microfluidic device was successfully applied for the determination of parabens in urine samples with relative recoveries between 86 and 100% for all analytes. Detection limits and quantitation limits were between 0.011-0.093 and 0.31-0.38 µg mL-1, respectively. Relative standard deviation was below 7% for all analytes. Furthermore, the environmentally-friendly solvent (Ca:Ty 2:1) used as SLM offered the same advantages in terms of membrane stability allowing consecutive extractions. Results were compared with experiments previously conducted using conventional (polypropylene) membranes, observing that highly green microextraction systems based on natural and biodegradable materials have proven to be an attractive alternative in microfluidic systems.

Microfluidic liquid-phase microextraction based on natural deep eutectic solvents immobilized in agarose membranes.

In liquid-phase microextraction (LPME), the sample and the acceptor are separated by a synthetic organic solvent, which is immobilized in a porous polymeric membrane of polypropylene or polyvinylidene fluoride. The organic solvent serves as extraction phase, while the polymeric membrane serves as support membrane. The combination of extraction phase and support membrane is termed supported liquid membrane (SLM). In this paper, we developed for the first time fully green and biodegradable supported SLMs, based on natural deep eutectic solvents as extraction phase and agarose as support membrane. This highly green approach was developed and studied with sulfonamide pharmaceuticals as model analytes, and performance was compared with LPME using conventional SLMs. All experiments were conducted in a microfluidic device. Model analytes were extracted from acidic sample (pH1.0) and into alkaline acceptor (pH12.0). Both sample and acceptor were pumped at 1 µL min-1 into the microfluidic device, and the optimal SLM was based on 3 µL of coumarin and thymol (1:2 molar ratio) as the extraction phase. The proposed green microfluidic device was successfully applied for the determination of sulfonamides in urine samples with spiking recoveries in the range of 77-100%. LPME with deep eutectic solvent immobilized in agarose showed similar performance as with conventional SLMs. Thus, the data presented in this paper demonstrate that highly green microextraction systems may be developed in the future, based on natural solvents and biodegradable materials.

A microchip device based liquid-liquid-solid microextraction for the determination of permethrin and cypermethrin in water samples.

In this work, for the first time, a microchip device integrating liquid-liquid-solid phase microextraction is presented. As a novel approach to microchip systems, liquid-liquid-solid microextraction was performed in a sandwiched microchip device. The microchip device consisted of three poly(methyl methacrylate) layers along with a double "Y"-shaped microchannel. As the stationary phase, polyacrylonitrile-C18 was synthesized and immobilized in the upper channel, while the beneath channel was used as a reservoir for the stagnant volume ratio of sample-to-extraction solvent phase. In this way, analytes were extracted from an aqueous sample through an organic phase into the stationary phase. The analytes were finally desorbed with a minimum amount of acetonitrile as the desorption solvent. Permethrin and cypermethrin were selected as the model analytes for extraction and subsequent analysis by gas chromatography-flame ionization detection. Under optimum conditions (extraction solvent; n-hexane, sample -to-extraction solvent volume ratio; 2:1, extraction time; 20 min, desorption solvent; acetonitrile, desorption volume; 200 µL, and desorption time; 15 min) detection limits were 3.5 and 6.0 ng mL-1 for permethrin and cypermethrin, respectively. Relative standard deviations for intra- and inter-day reproducibility were below 8.3%. Device-to-device precision was in the range of 8.1-9.6%. The proposed microchip device was successfully applied to determine permethrin and cypermethrin in water samples with recoveries in the range of 73-96%.

A selective and efficient microfluidic method-based liquid phase microextraction for the determination of sulfonamides in urine samples.

Liquid phase microextraction (LPME) into a microfluidic has undergone great advances focused on downscaled and miniaturized devices. In this work, a microfluidic device was developed for the extraction of sulfonamides in order to accelerate the mass transfer and passive diffusion of the analytes from the donor phase to the acceptor phase. The subsequent analysis was carried out by high performance liquid chromatography with UV-DAD (HPLC-DAD). Several parameters affecting the extraction efficiency of the method such as the supported liquid membrane, composition of donor and acceptor phase and flow rate were investigated and optimized. Tributyl phosphate was found to be a good supported liquid membrane which confers not only great affinity for analytes but also long-term stability, allowing more than 20 consecutive extractions without carry over effect. Under optimum conditions, extraction efficiencies were over 96 % for all sulfonamides after 10 minutes extraction and only 10 µL of sample was required. Relative standard deviation was between 3-5 % for all compounds. Method detection limits were 45, 57, 54 and 33 ng mL-1 for sulfadiazine (SDI), sulfamerazine (SMR), sulfamethazine (SMT) and sulfamethoxazole (SMX), respectively. Quantitation limits were 0.15, 0.19, 0.18 and 0.11 µg mL-1 for SDI, SMR, SMT SMX, respectively. The proposed microfluidic device was successfully applied for the determination of sulfonamides in urine samples with extraction efficiencies within the range of 86-106 %. The proposed method improves the procedures proposed to date for the determination of sulfonamides in terms of efficiency, reduction of the sample volume and extraction time.

A novel nanocomposite based on covalent organic polymer and nanocellulose for thin-film microextraction of imipramine from biological samples.

A novel covalent organic polymer was prepared using 1,5-diaminonaphthalene as a linker and cyanuric chloride as a node. A thin-film nanocomposite of 1,5-diaminonaphthalene covalent organic polymer and cellulose nanocrystalline was then fabricated via filtering and casting method. The effect of incorporation of various amounts of 1,5-diaminonaphthalene covalent organic polymer and cellulose nanocrystalline was studied to obtain an efficient nanocomposite thin-film with a large number of polar functional groups and high mechanical stability. Field emission scanning electron microscopy, X-ray diffraction, Fourier transform infrared spectrometry, and thermogravimetric analysis techniques were applied for the characterization of physicochemical properties of the prepared materials. Imipramine was determined in the biological samples using thin-film microextraction followed by gas chromatography flame ionization detection. Parameters affecting the extraction efficiency of imipramine were investigated. Under the optimized conditions, the limit of detection was 0.5 ng/mL. Film-to-film reproducibility for three different films fabricated under the same conditions (at three concentration levels) varied between 8.9 and 9.7%. The linear dynamic range covered more than three orders of magnitude (2-5000 ng/mL) with a determination coefficient of 0.9985. The method was successfully applied for preconcentration and determination of imipramine in biological samples with spiking recoveries between 78 and 93%.

A silica-based three-dimensional molecularly imprinted coating for the selective solid-phase microextraction of difenoconazole from wheat and fruits samples.

In this work, a selective silica-based molecular imprinted solid-phase microextraction (SPME) sorbent was prepared through the sol-gel process. Difenoconazole was used as a template to prepare imprinted materials on the surface of mesoporous silica. The SPME fiber followed by gas chromatography-electron capture detection was applied for the extraction and determination of difenoconazole. Fourier transform infrared spectrometry, field emission scanning electron microscopy, X-ray diffraction, and thermal gravimetry were used to characterize the imprinted SPME fiber. Two different procedures were presented to prepare MCM-41@SiO2-difenoconazole. Also, a non-MCM molecularly imprinted polymer was synthesized to investigate the effect of MCM-41 on the selectivity and extraction efficiency of the sorbent. The important parameters (i.e., desorption time and temperature, ionic strength, stirring rate, pH, extraction temperature and time) affecting the extraction performance of the method were optimized. Under optimum conditions, the limits of detection and quantification were found to be 0.002 and 0.005 ng mL-1, respectively. Linear dynamic range was in the range of 0.01-1 ng mL-1. The intra- and inter-day relative standard deviations and fiber-to-fiber reproducibility were in the range of 4.3-7.5, 3.3-8.2 and 7.4-9.7%, respectively. The fiber was successfully applied for the selective extraction of difenoconazole from wheat and fruit samples and satisfactory results with extraction recoveries >73% were obtained.

A solid-phase microextraction platinized stainless steel fiber coated with a multiwalled carbon nanotube-polyaniline nanocomposite film for the extraction of thymol and carvacrol in medicinal plants and honey.

A mechanically hard and cohesive porous fiber, with large surface area, for more strong attachment of the coating was provided by platinizing a stainless steel wire. Then, the platinized stainless steel fiber was coated with a multiwalled carbon nanotube/polyaniline (MWCNT/PANI) nanocomposite using electrophoretic deposition (EPD) method and applied for the extraction of thymol and carvacrol with direct-immersion solid-phase microextraction (DI-SPME) method followed by high-performance liquid chromatography-ultraviolet detection (HPLC-UV) quantification. To provide a larger coarse surface for the tightened attachment of coating on the fiber, a stainless steel wire was platinized using a suitable optimized EPD method. Different experimental parameters were studied and the optimal conditions were obtained as: pH of the sample solution: 2; extraction time: 60min; salt content in the sample solution: 1% w/v NaNO3; desorption time: 60min; type and volume of the desorption solvent: acetonitrile, 100µL. Under the optimized conditions, limits of detection (LODs) were 0.6 and 0.8µgmL(-1) for thymol and carvacrol, respectively. Linear dynamic range (LDR) for the calibration curves of both analytes were 1-80µgmL(-1). Relative standard deviation (RSD%, n=6) was 6.8 for thymol and 12.7 for carvacrol. The proposed fiber was successfully applied for the recovery and determination of thymol and carvacrol in thyme, savory, and honey samples.