A closer look at how the dispersive liquid-liquid microextraction method works. Investigation of the effect of solvent mixture composition on the quality and stability of the cloudy state.

The dispersive liquid-liquid microextraction (DLLME) is one of the most popular miniaturized extraction procedures. In this paper, the degree of dispersion and dispersion stability were studied with the aim to assess the correlations of these parameters with efficiency for the selected analytical application. The dependence between the degree of dispersion (cloudy state quality) and its stability obtained by various emulsification procedures, such as solvent-assisted emulsification (using various dispersive solvents) and mechanical emulsification (using auxiliary energies), is investigated and discussed. It was found out that the degree of dispersion depends on the type of emulsification procedure and decreases in the series: solvent-assisted (SA-) = ultrasound-assisted (UA-) > air-assisted (AA-) > vortex-assisted (VA-) emulsification. The emulsion stability depends on the degree of dispersion and there were 1810 and 2070 s for the most effective emulsification procedures, such us solvent-assisted and ultrasound-assisted emulsification, respectively. A comparison between the sensitivity of the analytical methods (using spectrophotometric determination of the anionic surfactants) and the degree of dispersion have been made. The sensitivity of the methods was ranked as follows: DLLME > UA-LLME > VA-LLME > AA-LLME.

Deep eutectic solvent-based shaking-assisted extraction for determination of bioactive compounds from Norway spruce roots.

Polyphenolic compounds play an essential role in plant growth, reproduction, and defense mechanisms against pathogens and environmental stresses. Extracting these compounds is the initial step in assessing phytochemical changes, where the choice of extraction method significantly influences the extracted analytes. However, due to environmental factors, analyzing numerous samples is necessary for statistically significant results, often leading to the use of harmful organic solvents for extraction. Therefore, in this study, a novel DES-based shaking-assisted extraction procedure for the separation of polyphenolic compounds from plant samples followed by LC-ESI-QTOF-MS analysis was developed. The DES was prepared from choline chloride (ChCl) as the hydrogen bond acceptor (HBA) and fructose (Fru) as the hydrogen bond donor (HBD) at various molar ratios with the addition of 30% water to reduce viscosity. Several experimental variables affecting extraction efficiency were studied and optimized using one-variable-at-a-time (OVAT) and confirmed by response surface design (RS). Nearly the same experimental conditions were obtained using both optimization methods and were set as follows: 30 mg of sample, 300 mg of ChCl:Fru 1:2 DES containing 30% w/w of water, 500 rpm shaking speed, 30 min extraction time, 10°C extraction temperature. The results were compared with those obtained using conventional solvents, such as ethanol, methanol and water, whereby the DES-based shaking-assisted extraction method showed a higher efficiency than the classical procedures. The greenness of the developed method was compared with the greenness of existing procedures for the extraction of polyphenolic substances from solid plant samples using the complementary green analytical procedure index (ComplexGAPI) approach, while the results for the developed method were better or comparable to the existing ones. In addition, the practicability of the developed procedure was evaluated by application of the blue applicability grade index (BAGI) metric. The developed procedure was applied to the determination of spruce root samples with satisfactory results and has the potential for use in the analysis of similar plant samples.

Innovative approaches in cloud-point extraction.

Cloud-point extraction (CPE) is a pre-treatment technique for the extraction and preconcentration of different chemical compounds, such as metal ions, pesticides, drugs, phenols, vitamins etc., from various samples. CPE is based on the phenomenon of two phases (micellar and aqueous) forming after the heating of an aqueous isotropic solution of a non-ionic or zwitterionic surfactant above the cloud-point temperature. If analytes are added to the surfactant solution under suitable conditions, they should be extracted into the micellar phase, also called the surfactant-rich phase. Recently, the traditional CPE procedure is being increasingly replaced by improved CPE procedures. In this study, recent advances in CPE over the last three years (2020 - 2022), including the application of various innovative approaches, are reviewed. In addition to the basic principle of CPE, alternative extraction media in CPE, CPE supported by various auxiliary energies, a different modified CPE procedure and the use nanomaterials and solid-phase extraction in combination with CPE are presented and discussed. Finally, some future trends for improved CPE are presented.

Deep Eutectic Solvents or Eutectic Mixtures? Characterization of Tetrabutylammonium Bromide and Nonanoic Acid Mixtures.

Deep eutectic solvents have quickly attracted the attention of researchers because they better meet the requirements of green chemistry and thus have the potential to replace conventional hazardous organic solvents in some areas. To better understand the nature of these mixtures, as well as expand the possibilities of their use in different industries, a detailed examination of their physical properties, such as density, viscosity, the nature of the interactions between their constituents, the phase diagrams, depression of their melting point, and interpretation of these results is necessary. In this work, the mixtures of tetrabutylammonium bromide (TBAB) and nonanoic acid (NA) in different molar ratios are theoretically and experimentally investigated by applying a phase diagram constructed on the basis of differential scanning calorimetry measurements and COSMO-RS model. Spectral properties are investigated based on Fourier transform infrared spectroscopy and density functional theory. The observed eutectic point indicates the formation of a DES in the TBAB-NA system in a 1:2 molar ratio. This is due to the presence of hydrogen bonds between the carboxyl group from the NA molecule and the bromine atom from the TBAB molecule. Other eutectic mixtures are most likely the solutions of TBAB in NA, in which hydrogen bonds predominate between acid molecules.

Application of liquid-phase microextraction to the analysis of plant and herbal samples.

INTRODUCTION: The analysis of plant and herbal samples is a challenging task for analytical chemists due to the complexity of the matrix combined with the low concentration of analytes. In recent years different liquid-phase microextraction (LPME) techniques coupled with a variety of analytical equipment have been developed for the determination of both organic and inorganic analytes. OBJECTIVE: Over the past few years, the number of research papers in this field has shown a markedly growing tendency. Therefore, the purpose of this review paper is to summarise and critically evaluate research articles focused on the application of LPME techniques for the analysis of plant and herbal samples. RESULTS: Due to the complex nature of the samples, the direct application of LPME techniques to the analysis of plants has not often been done. LPME techniques as well as their modalities have been commonly applied in combination with other pretreatment techniques, including a solid-liquid extraction technique supported by mechanical agitation or auxiliary energies for plant analysis. Applications and the most important parameters are summarised in the tables. CONCLUSION: This review summarises the application of the LPME procedure and shows the major benefits of LPME, such as the low volume of solvents used, high enrichment factor, simplicity of operation and wide selection of applicable detection techniques. We can expect further development of microextraction analytical methods that focus on direct sample analysis with the application of green extraction solvents while fully automating procedures for the analysis of plant materials.

In situ decomposition of deep eutectic solvent as a novel approach in liquid-liquid microextraction.

In this study, a novel approach for effective liquid-liquid microextraction based on deep eutectic solvent (DES) decomposition was suggested for the first time. It was established that DESs synthesized from tetrabutylammonium bromide and long-chain alcohols decomposed in aqueous phase resulting in in situ dispersion of organic phase and extraction of hydrophobic analyte(s). It this process long-chain alcohol acted as an extraction solvent and tetrabutylammonium bromide acted as a dispersive agent and promoted mass transfer between aqueous and organic phases as a salting out agent. Phenomenon of DES decomposition was studied in detail and applied for separation and preconcentration in chemical analysis for the first time. The developed approach was applied for 17ß-estradiol microextraction from transdermal gel samples as a proof-of-concept example. The results showed that the in situ dispersed organic phase obtained can provide efficient extraction of 17ß-estradiol with good extraction recovery (95 ±â€¯5%) and excellent reproducibility (6%). The reported approach proves to be fast, simple, and inexpensive.

Liquid Phase and Microwave-Assisted Extractions for Multicomponent Phenolic Pattern Determination of Five Romanian Galium Species Coupled with Bioassays.

Background: Galium is a plant rich in iridoid glycosides, flavonoids, anthraquinones, and small amounts of essential oils and vitamin C. Recent works showed the antibacterial, antifungal, antiparasitic, and antioxidant activity of this plant genus. Methods: For the determination of the multicomponent phenolic pattern, liquid phase microextraction procedures were applied, combined with HPLC-PDA instrument configuration in five Galium species aerial parts (G. verum, G. album, G. rivale, G. pseudoaristatum, and G. purpureum). Dispersive Liquid⁻Liquid MicroExtraction (DLLME) with NaCl and NAtural Deep Eutectic Solvent (NADES) medium and Ultrasound-Assisted (UA)-DLLME with ß-cyclodextrin medium were optimized. Results: The optimal DLLME conditions were found to be: 10 mg of the sample, 10% NaCl, 15% NADES or 1% ß-cyclodextrin as extraction solvent-400 µL of ethyl acetate as dispersive solvent-300 µL of ethanol, vortex time-30 s, extraction time-1 min, centrifugation at 12000× g for 5 min. Conclusions: These results were compared with microwave-assisted extraction procedures. G. purpureum and G. verum extracts showed the highest total phenolic and flavonoid content, respectively. The most potent extract in terms of antioxidant capacity was obtained from G. purpureum, whereas the extract obtained from G. album exhibited the strongest inhibitory effect against tyrosinase.

Use of sequential injection analysis with lab-at-valve and an optical probe for simultaneous spectrophotometric determination of ascorbic acid and cysteine by mean centering of ratio kinetic profiles.

Two new, simple, relatively fast and robust methods for the simultaneous kinetic determination of binary mixtures of ascorbic acid (Asc) and cysteine (Cys) were developed using the mean centering of ratio kinetic profiles method. The methods are based on the difference in the reaction rates of Asc and Cys with 18-molybdodiphosphate at pH 5.1. An optical probe as well as the sequential injection analysis lab-at-valve (SIA-LAV) method were used to carry out simultaneous kinetic analysis. The benefits of the mean centering of ratio kinetic profiles method were shown in comparison with other spectrophotometric kinetic methods. Asc and Cys can be determined in the concentration ranges 20-200 and 8-90 µmol L-1 with the batch spectrophotometric method and 10-200 and 4-40 µmol L-1 with the SI-LAV method, respectively. The method was successfully used to determine Asc and Cys in dietary supplements.

Development of novel techniques to extract phenolic compounds from Romanian cultivars of Prunus domestica L. and their biological properties.

In the present work, fourteen cultivars of Prunus domestica were analyzed to investigate their phenolic pattern with the purpose of using the leaves as potential resources of bioactive compounds in the pharmaceutical and food industry. Microwave-assisted extraction (MAE), dispersive liquid-liquid microextraction and sugaring-out liquid-liquid extraction techniques were optimized in order to obtain an exhaustive multi-component panel of phenolic compounds. The best phenolic-enriched recovery was achieved using MAE in water:methanol (30:70), and this procedure was further applied for quantitative analysis of phenolic compounds in real samples. In order to prove the safeness of these extracts, the biological potential of the Prunus cultivars was tested by several in vitro antioxidant and enzyme inhibitory assays. Moreover, their cytotoxicity was evaluated on human gingival fibroblasts (HGFs), and in most of the cases the treatment with different concentrations of extracts didn't show cytotoxicity up to 500 µg/mL. Only 'Carpatin' and 'Minerva' cultivars, at 250 and 500 µg/mL, reduced partially cell viability of HGFs population. Noteworthy, Centenar cultivar was the most active for the α-glucosidase inhibition (6.77 mmolACAE/g extract), whereas Ialomița cultivar showed the best antityrosinase activity (23.07 mgKAE/g extract). Overall, leaves of P. domestica represent a rich alternative source of bioactive compounds.

Simultaneous determination of rutin and ascorbic acid in a sequential injection lab-at-valve system.

A green, simple, accurate and highly sensitive sequential injection lab-at-valve procedure has been developed for the simultaneous determination of ascorbic acid (Asc) and rutin using 18-molybdo-2-phosphate Wells-Dawson heteropoly anion (18-MPA). The method is based on the dependence of the reaction rate between 18-MPA and reducing agents on the solution pH. Only Asc is capable of interacting with 18-MPA at pH 4.7, while at pH 7.4 the reaction with both Asc and rutin proceeds simultaneously. In order to improve the precision and sensitivity of the analysis, to minimize reagent consumption and to remove the Schlieren effect, the manifold for the sequential injection analysis was supplemented with external reaction chamber, and the reaction mixture was segmented. By the reduction of 18-MPA with reducing agents one- and two-electron heteropoly blues are formed. The fraction of one-electron heteropoly blue increases at low concentrations of the reducer. Measurement of the absorbance at a wavelength corresponding to the isobestic point allows strictly linear calibration graphs to be obtained. The calibration curves were linear in the concentration ranges of 0.3-24mgL-1 and 0.2-14mgL-1 with detection limits of 0.13mgL-1 and 0.09mgL-1 for rutin and Asc, respectively. The determination of rutin was possible in the presence of up to a 20-fold molar excess of Asc. The method was applied to the determination of Asc and rutin in ascorutin tablets with acceptable accuracy and precision (1-2%).

Use of Innovative (Micro)Extraction Techniques to Characterise Harpagophytum procumbens Root and its Commercial Food Supplements.

INTRODUCTION: For the determination of harpagoside and the wide phenolic pattern in Harpagophytum procumbens root and its commercial food supplements, dispersive liquid-liquid microextraction (DLLME), ultrasound-assisted DLLME (UA-DLLME), and sugaring-out liquid-liquid extraction (SULLE) were tested and compared. OBJECTIVES: In order to optimise the extraction efficiency, DLLME and UA-DLLME were performed in different solvents (water and aqueous solutions of glucose, ß-cyclodextrin, (2-hydroxypropyl)-ß-cyclodextrin, sodium chloride, natural deep eutectic solvent, and ionic liquid). MATERIAL AND METHODS: The plant material was ground and sieved to obtain a uniform granulometry before extraction. Commercial food supplements, containing H. procumbens are commercially available in Italy. RESULTS: The most effective sodium chloride-aided-DLLME was then optimised and applied for analyses followed by HPLC-PDA. For comparison, microwave-assisted extraction was performed using the same solvents and the best results were obtained using 1% of ß-cyclodextrin or 15% of sodium chloride. CONCLUSION: All commercial samples respected the European Pharmacopoeia monograph for this plant material, showing a harpagoside content ≥ 1.2%. Copyright © 2017 John Wiley & Sons, Ltd.

Flow method based on liquid-liquid extraction using deep eutectic solvent for the spectrofluorimetric determination of procainamide in human saliva.

In the current study, liquid-liquid extraction, using deep eutectic solvent (DES) as a "green" extraction solvent, was coupled with a stepwise injection system for the first time. The suggested approach was applied for the development of spectrofluorimetric method for procainamide determination. The method is based on aspiration of saliva sample and DES (choline chloride with glycerol at a 1:2M ratio) solution into the mixing chamber of a flow system, followed by injection of acetonitrile into the mixed DES-sample solution. The extraction process and final phase separation were then promoted by air-bubbling. After phase separation, the DES phase, containing the extracted procainamide, was transported to a spectrofluorimetric detector. The excitation and emission wavelengths were set at 280nm and 347nm, respectively. The calibration plot was linear in the range of 5×10-6 to 5×10-5molL-1. The limit of detection, calculated as 3σ of a blank test (n=10), was found to be 1.5×10-6molL-1. The developed method was successfully applied for the determination of procainamide in human saliva samples, and the analytical results agreed rather well with the results obtained by the reference HPLC-UV method.

Automated alkaline-induced salting-out homogeneous liquid-liquid extraction coupled with in-line organic-phase detection by an optical probe for the determination of diclofenac.

A fully automated alkaline-induced salting-out homogeneous liquid-liquid extraction (AI-SHLLE) procedure coupled with in-line organic-phase detection by an optical probe has been suggested. Diclofenac was used as a proof-of-concept analyte. The method is based on the oxidation of diclofenac with potassium ferricyanide in an alkaline medium followed by separation of the acetonitrile phase from the homogeneous sample solution and simultaneous extraction of the derivative. Sodium hydroxide serves as both the alkaline agent for the derivatization of diclofenac and as the salting-out agent for the acetonitrile-rich phase formation. Absorbance of the derivative in the acetonitrile-rich phase was measured in-line using an optical probe. The calibration graph was linear over the range of 2.5-60µmolL-1 with the regression coefficient equal to 0.9997. The LOD calculated from the calibration plot based on 3σ was 0.8µmolL-1. The sample throughput was 7 samplesh-1. The method was applied for the determination of diclofenac in spiked saliva samples and pharmaceutical preparations and the results were compared with those obtained by the reference method.

Automated sugaring-out liquid-liquid extraction based on flow system coupled with HPLC-UV for the determination of procainamide in urine.

A fully automated sugaring-out assisted liquid-liquid extraction procedure was suggested. The procedure was based on the separation of the acetonitrile phase, containing a target analyte from the homogeneous sample solution after injection of sugaring-out reagent (glucose) into a mixing chamber of the flow system. Air bubbling was used to promote the extraction process and phase separation. After the fast phase separation in the mixing chamber, the acetonitrile phase containing the target analyte was transferred to an HPLC-UV system. Under the optimal conditions, the detector response of procainamide was linear in the concentration range of 6×10-7-4×10-5molL-1. The limit of detection, calculated from a blank test based on 3σ, was 2×10-7molL-1. The proposed method was successfully applied for the determination of procainamide in human urine samples and the analytical results agreed fairly well with the results obtained by reference CE method.

Using an Optical Probe as the Microdrop Holder in Headspace Single Drop Microextraction: Determination of Sulfite in Food Samples.

A novel headspace single-drop microextraction method (HS-SDME) for determination of sulfite in the form of sulfur dioxide was developed. An optical probe was used as the droplet holder in the HS-SDME procedure, and the analytical signal (absorbance) was monitored online during the extraction process. The method is based on the conversion of sulfite to volatile sulfur dioxide by acidification of the analyzed solution. The liberated SO2 was absorbed by 25 µL of an aqueous mixed reagent solution placed on the optical probe tip and containing Fe(III), 1,10-phenantroline, and an acetic buffer solution of pH 5.6. During the extraction process, Fe(III) reduces to Fe(II) and the Fe(II) formed then reacts with 1,10-phenantroline to form a colored complex. Absorbance was measured at 510 nm. The calibration plot was linear in the range 0.032-0.320 mg L-1 of sulfite (as SO2), with a correlation coefficient of 0.9989. The limit of detection (LOD), calculated as three times the standard deviation of the blank test (n = 10), was found to be 8 µg L-1. The method was applied for analysis of real food samples, such as wine, jam, and juice.

Interfacial reaction using particle-immobilized reagents in a fluidized reactor. Determination of glycerol in biodiesel.

A novel fluidized beads strategy for utilization of particle-immobilized reagents in flow analysis was developed in this study. The performance of the suggested strategy was demonstrated by the determination of glycerol in biodiesel. This analytical task was used as a proof-of-concept example. The method is based on on-line extraction of glycerol from biodiesel into aqueous stationary phase of extraction-chromatographic column, followed by elution and spectrophotometric determination in the form of copper glycerate formed in a fluidized reactor of stepwise injection system. The floating of cation exchange resin Dowex(®) 50WX4, saturated with Cu(II) ions in liquid phase, was accomplished by air-bubbling. The linear range was from 100 to 1000 mg kg(-1), and the limit of detection, calculated as 3s of a blank test (n = 5), was found to be 30 mg kg(-1). The method was successfully applied to the analysis of biodiesel and biodiesel-blend (B 20) samples.

A novel vortex-assisted liquid-liquid microextraction approach using auxiliary solvent: Determination of iodide in mineral water samples.

A novel vortex-assisted liquid-liquid microextraction (VA-LLME) for determination of iodide was developed. The method includes the oxidation of iodide with iodate in the presence of hydrochloric acid followed by VA-LLME of the ion-pair formed between ICl2(-) and Astra Phloxine reagent (AP) and subsequent absorbance measurement at 555nm. The appropriate experimental conditions were investigated and found to be: 5mL of sample, 0.27molL(-)(1) HCl, 0.027mmolL(-1) KIO3 as the oxidation agent, 250µL of extraction mixture containing amyl acetate as the extraction solvent and carbon tetrachloride as the auxiliary solvent (1:1, v/v), 0.04mmolL(-1) AP reagent, vortex time: 20s at 3000rpm, centrifugation: 4min at 3000rpm. The calibration plot was linear in the range 16.9-169µg L(-1) of iodide, with a correlation coefficient (R(2)) of 0.996, and the relative standard deviation ranged from 1.9 to 5.7%. The limit of detection (LOD) and limit of quantification (LOQ) were 1.75 and 6.01µgL(-)(1) of iodide, respectively. The suggested procedure was applied for determination of iodide in real mineral water samples.

A fully automated effervescence-assisted switchable solvent-based liquid phase microextraction procedure: Liquid chromatographic determination of ofloxacin in human urine samples.

A novel fully automated effervescence-assisted switchable solvent-based liquid phase microextraction procedure has been suggested. In this extraction method, medium-chain saturated fatty acids were investigated as switchable hydrophilicity solvents. The conversion of fatty acid into hydrophilic form was carried out in the presence of sodium carbonate. The injection of sulfuric acid into the solution decreased the pH value of the solution, thus, microdroplets of the fatty acid were generated. Carbon dioxide bubbles were generated in-situ, and promoted the extraction process and final phase separation. The performance of the suggested approach was demonstrated by the determination of ofloxacin in human urine samples using high-performance liquid chromatography with fluorescence detection. This analytical task was used as a proof-of-concept example. Under the optimal conditions, the detector response of ofloxacin was linear in the concentration ranges of 3·10(-8)-3·10(-6) mol L(-1). The limit of detection, calculated from a blank test based on 3σ, was 1·10(-8) mol L(-1). The results demonstrated that the presented approach is highly cost-effective, simple, rapid and environmentally friendly.

A fully automated effervescence assisted dispersive liquid-liquid microextraction based on a stepwise injection system. Determination of antipyrine in saliva samples.

A first attempt to automate the effervescence assisted dispersive liquid-liquid microextraction (EA-DLLME) has been reported. The method is based on the aspiration of a sample and all required aqueous reagents into the stepwise injection analysis (SWIA) manifold, followed by simultaneous counterflow injection of the extraction solvent (dichloromethane), the mixture of the effervescence agent (0.5 mol L(-1) Na2CO3) and the proton donor solution (1 mol L(-1) CH3COOH). Formation of carbon dioxide microbubbles generated in situ leads to the dispersion of the extraction solvent in the whole aqueous sample and extraction of the analyte into organic phase. Unlike the conventional DLLME, in the case of EA-DLLME, the addition of dispersive solvent, as well as, time consuming centrifugation step for disruption of the cloudy state is avoided. The phase separation was achieved by gentle bubbling of nitrogen stream (2 mL min(-1) during 2 min). The performance of the suggested approach is demonstrated by determination of antipyrine in saliva samples. The procedure is based on the derivatization of antipyrine by nitrite-ion followed by EA-DLLME of 4-nitrosoantipyrine and subsequent UV-Vis detection using SWIA manifold. The absorbance of the yellow-colored extract at the wavelength of 345 nm obeys Beer's law in the range of 1.5-100 µmol L(-1) of antipyrine in saliva. The LOD, calculated from a blank test based on 3σ, was 0.5 µmol L(-1).

Vibrational spectroscopic study of dehydroacetic acid and its cinnamoyl pyrone derivatives.

The infrared and Raman spectra of dehydroacetic acid and some of its derivatives were measured. The assignments of the vibrational bands were based on quantum chemical calculations and normal coordinate analysis. The optimized structures, atomic net charges and dipole moments of the investigated molecules were also results of our quantum chemical calculations. The analysis of the last properties made possible a deeper insight into the structure and substituent effect on the investigated molecules. One of them is presented in the graphical abstract.