Micellar Extraction Research Articles

Analysis of ionic and nonionic surfactants blends used for the reverse micellar extraction of Lactoperoxidase from whey

AbstractBovine Lactoperoxidase (LP), a minor whey protein, is used as an antimicrobial in cosmetic, food, and pharmaceutical preparations. Industries are in pursuit of reliable, cheap, and scalable purification methods as the conventional techniques for LP purification like chromatography and membrane separation suffer from several drawbacks. The present work investigates the selective reverse micellar extraction of LP using the reverse micellar system formed by mixing food grade nonionic (Tween, Span, and Triton series) and ionic (AOT) surfactant blends. The analysis of LP extraction efficiency was performed by varying the concentration of nonionic surfactants with a constant AOT concentration of 100 mM and the initial pH of the system. Complete LP solubilization was achieved with reverse micelles formed by 100 mM AOT and 20 mM Tween 80 at pH 8. It was found that the extraction efficiency was dependent on the chain length or the number of ethylene oxide units in the Triton surfactant tail and the carbon–carbon double bond in Tween 80 tail, that is, on oleic acid. Span series however showed poor extraction in the organic phase substantiating the lesser water content. The forward extracted LP was successfully back‐extracted into a fresh aqueous phase containing 1 M KCl at pH 10.5. The aqueous phase (whey) from the forward transfer can be further used to fractionate other whey proteins.

Micellar extraction with vesicle coated multi-walled carbon nanotubes to assist the dispersive micro-solid-phase extraction of natural phenols in Dendrobium

Here, catanionic surfactant vesicles were prepared by varying the types and compositions of anions and cations and the number of alkyl tails of the surfactants. The formed vesicles were employed to disaggregate and stabilize multiwalled carbon nanotubes bundles in aqueous solutions. Furthermore, the vesicle coated carbon nanotubes were used as the adsorbent in the dispersive micro-solid phase extraction. Additionally, micellar extraction was employed for the sample pre-extraction to avoid the use of toxic organic extraction solvents. The relative parameters that affect the extraction efficiency of targets were optimized using response surface methodology. Under the optimal microextraction conditions, the analytical performance of the established method was evaluated. The limits of detection (2.3−13 ng/mL) and quantification (7.6−42 ng/mL), inter- and intra- day precision (1.2–4.0 %, 2.0–5.0 %), and spiked recovery values (80–91 %) were obtained. The proposed method showed high sensitivity, precision and trueness. It was successfully applied to analyze phenols in Dendrobium genus samples.

Micellar Extraction Preconcentration and Colorimetric Determination of Some Phenols

We studied the extraction and preconcentration of phenol and its hydroxy derivatives using individual and mixed micelles of nonionic and cationic surfactants with the digital recording of the analytical signal. This approach is based on the color reaction of phenols with diazotized 4-nitroaniline and micellar extraction of the products of their interaction (azo compound). The micellar extraction parameters (pH, the concentration of reagents, NaOH, and ethanol) were optimized for the efficient extraction of azo compounds of the studied phenols. The proposed method enables the determination of phenol and resorcinol at the MPC level with an error not exceeding 10%. For a qualitative assessment of the presence of phenols, the geometric parameters (area S, perimeter P) of the corresponding radar plots were used along with colorimetric parameters (R, G, B). The calibration dependences are linear within 1 × 10–7–3 × 10–5 M for phenol and 1 × 10–7–2 × 10–5 M for resorcinol and phloroglucinol. The developed colorimetric method was tested in the determination of resorcinol in the Rezortsinol pharmaceutical preparation.

PREPARATION, CHARACTERIZATION, AND FORMULATION OF SOLID LIPID NANOPARTICLES LOTION FROM MULBERRY ROOTS (MORUS ALBA L.)

Objective: Tween 80 has been used as a solvent for the extraction of phenolic compounds because this surfactant has both hydrophilic and hydrophobicproperties. Solid lipid nanoparticles (SLNs) have been developed to improve penetration through the skin layer. We investigated the efficacy of usingthe microwave-assisted micellar extraction (MAME) approach for extracting oxyresveratrol from Morus alba roots and also to develop an SLN lotion.Methods: The M. alba roots were extracted with Tween 80 in a microwave for 18 min, and the extract was used to develop SLN with differentconcentrations of glyceryl monostearate. The SLNs from M. alba root extracts were prepared by a high-speed homogenization technique (25,000 rpmfor 15 min). The SLNs produced were characterized as per particle size, polydispersity index (PDI), and zeta potential. The SLNs with the bestcharacteristics were used to formulate a lotion using a high-pressure homogenizer.Results: Extraction using MAME showed improved extraction efficiency. The oxyresveratrol concentration from the extract was 2.77%. The SLN with2.5% glyceryl monostearate showed the optimum result, with a particle size of 130.20 nm, a PDI of 0.278, and a zeta potential of −21.8 mV. The SLNlotion exhibited a particle size of 285.9 nm and a PDI of 0.360. The SLN lotion also had good penetration, with a flux of 4.70 μg cm−2/h.Conclusion: MAME is an efficient method for extracting oxyresveratrol from M. alba roots. The SLN with 2.5% glyceryl monostearate exhibited theoptimum characteristics, and the SLN lotion showed good characteristics, including skin penetration.

Reverse micellar extraction of dyes based on fatty acids and recoverable organic solvents

Reverse micelle extraction occurs when a substrate is removed from the aqueous phase to an organic phase by reverse micelles in an immiscible solvent with water. In this work, three fatty acids: stearic acid (C18), palmitic acid (C16) and lauric acid (C12) were investigated in the extraction of the methylene blue (MB+) dye, as a model of cationic compound, from aqueous solutions. Experiments were performed in quartz cuvettes with 2.0 mL of MB+ 6.25 x 10-4 mol L-1 at pH 10, and 1.0 mL of surfactants in isopentanol, from 3.0 to 6.0 mmol L-1. At pH 10, the three fatty acids presented efficiency in MB+ removal superior to 99%. However, for all tested conditions, C18 at 3.7 mmol L-1 presented the fastest separation of phases. By chemometric factorial design, the maximum efficiency of separation was superior to 99% and occurred with at least 21% of organic phase, the most important variable for this system. In optimal condition, the total time of separation of phases was around 5h. After the forward process, the organic phase containing reverse micelles and MB+ was acidified with diluted H2SO4 for protonation and precipitation of fatty acids. Therefore, in this case, reverse micelles were disrupted and MB+ was concentrated in a small volume of acid aqueous solution, thus allowing around 90% of recuperation of isopentanol for reuse. The main advantages of this process include selectivity, mild thermal conditions, low energy consumption, potential for use at the industrial scale and possibility of application in the removal of other cationic pollutants.

Response surface optimization of the forward extraction of jacalin from jackfruit seeds using AOT/isooctane reverse micellar system

Jacalin is the major protein contained in the crude extract of jackfruit (Artocarpus heterophyllus) seed that specifically recognizes and binds reversibly to galactose. Conventionally, purification of jacalin is carried out using the tedious and costly chromatographic techniques. In this study, extraction of jacalin from jackfruit seed crude extract were done using the sodium bis(2-ethylhexyl) sulfosuccinate (AOT)-based reverse micellar system. Reverse micellar extraction is an attractive alternative for downstream processing of various proteins. A successful reverse micellar extraction consists of two basic steps: forward and backward extraction. Forward extraction transfers a target protein from an aqueous solution into the reverse micellar solution, while backward extraction releases the protein from the reverse micelles structure into a new aqueous solution. The effects of the aqueous phase pH, NaCl concentration and AOT concentration on the forward extraction efficiency (FEE) are investigated using the response surface methodology (Box-Behnken Design). The main effects and interactions of the parameters are analyzed through the 3D surface plots. The optimum conditions for forward extraction were determined as follows: aqueous phase pH 4.58, 125 mM NaCl and 40 mM AOT. Under the optimal conditions, the FEE reached 88.04±1.30%, closer to 87.99% predicted by the model. The results indicated that AOT/isooctane reverse micelle system is effective in extracting jacalin from the jackfruit seed crude extract and verified the practicability of the BBD model for optimizing the main parameters in the forward extraction of jacalin.

Міцелярна екстракція катіонних ПАР та їх іонних асоціатів з додецилсульфат-аніоном фазами неіонної ПАР при температурі помутніння

The distribution of individual cationic surfactants and their ionic associates with the anionic surfactant in the cloud point extraction systems, based on non-ionic surfactant OP-7 was investigated. Determined that the dependence of the extraction degree of halides alkylpyridiniums on carboxylic change length in optimal conditions for extraction is characterized by the existence of maximum for cationic surfactant where n = 12. Decrease as well as increase in carboxylic change length of cation of alkylpyridiniums diminishes the effectiveness of their extraction into the surfactant-rich phase. The peculiarities of inter phase distribution of ionic associates cationic surfactant – anionic surfactant in the micellar extraction systems are investigated. It was shown, that with the increase in carboxylic change length of cationic surfactant the dependence of extraction degree of associates with anionic surfactant is characterized by the existence of the maximum. Most effective into the surfactant-rich phase extracted of ionic associates of three- and tetradecylpyridinium. Decreasing and increasing of hydrophobicity of alkylpyridiniums diminishes the effectiveness of extraction of associates. The established effects are explained by the hydrophobic correspondence between the substrate and the surfactant-rich phase of non-ionic surfactant.

The Environmental Impact of Polycyclic Aromatic Hydrocarbons: Mechanism of Extraction by Bio-Surfactant in a Microwave

In N’Djamena, the use and marketing of certain hydrocarbons does not comply with any standard in force or with regulations provided for this purpose. Their evaporation and unregulated release into the wild significantly affects the ecosystem. The present work consists in developing a method of extraction from sediments polluted by bio surfactant (rhamnolipid) assisted by microwaves. The goal here is to look for the presence of polycyclic aromatic hydrocarbons in polluted sediments. The rhamnolipid used consists of monorhamnolipids and di-rhamnolipids, its emulsion index is 64.66% and is composed of saturated and unsaturated fatty acids having carbon numbers ranging from 12 to 18. We used the microwave micellar extraction process. It was made by keeping the concentration values fixed at 0.15 g/L and the power at 400 W and more. We observed a positive interaction of the rhamnolipid concentration factors and the power of the microwave to obtain the optimal conditions at the time of 50 S, at the concentration of 0.16 g/L and at the power of 443 W for a rate of optimal extraction of 0.91%. Chromatographic analysis by GC-MS of the optimum extracts allowed us to identify twelve (12) C10 to C43 n-alkanes and eight (8) PAHs. It emerges from this analysis that the rhamnolipid extracts seven (7) Polycyclic Aromatic Hydrocarbons in equivalent proportions while the tween 80 extracts only four (4) Polycyclic Aromatic Hydrocarbons with a high proportion (80.02%) of benzo [a] antracene.

Structural changes of a protein extract from apple with polyphenoloxidase activity obtained by cationic reversed micellar extraction induced by high-pressure carbon dioxide and thermosonication

Polyphenoloxidase from apple was extracted and further concentrated by cationic reversed micellar extraction. Previous to reversed micellar extraction a crude protein extract was obtained using AG2-X8 as adsorbent of phenolic compounds and the detergent Triton X-100. Forward and backward extraction conditions were optimized by using dodecyl trimethyl ammonium bromide as surfactant in the organic phase. Optimization was carried out to obtain the highest value of PPO activity recovery and the purification fold at the different experimental conditions. Under the optimum extraction conditions, PPO activity recovery was 99% and purification fold reached a value of 17, showing that reversed micellar extraction was a good technique as a first step to concentrate on a targeted enzyme. After removing some impurities by centrifuge ultrafiltration, the protein extract with PPO activity was treated by pressurized carbon dioxide and thermosonication achieving residual PPO activity values of 16 ± 3 and 9 ± 1%, respectively. Quenching experiments by iodide performed in the non-treated extract and in the treated extracts revealed conformational changes of this protein fraction reflected in the greater exposure of the fluorophore to the quencher.

Cationic reverse micellar based purification of recombinant glutaminase free L-asparaginase II of Bacillus subtilis WB800N from fermentation media

Reverse micellar extraction (RME), a liquid-liquid based separation is a versatile tool for protein purification. A statistical approach was employed for the purification of recombinant glutaminase free anti-cancerous enzyme viz., l-asparaginase II to evaluate the effects of RME in current study. The cationic system (CTAB/iso-octane/hexanol/butanol) was used in RME to optimize both forward and backward protein extraction efficiency. By adapting Taguchi's orthogonal array (OA), maximum forward extraction efficiency (FEE) of 86.98% with 84.82% enzyme activity recovery and 1.04 times purification fold achieved with the optimized parameters. Under the optimal levels, the back extraction efficiency (BEE) was observed to be 96.97% with 93.07% enzyme activity recovery and 1.38 times purification fold. Further, mass transfer kinetic studies of RME indicated the mass transfer coefficients of forward and backward extraction to be 0.049 min−1 and 0.036 min−1 respectively.

Fractional Factorial Analysis for Identifying Significant Factors in Extraction of Jacalin From Jackfruit Seeds Using Anionic Reverse Micellar System

Abstract Reverse micellar extraction is emerging as one of the most reliable method for extraction and purification of various proteins and enzymes. In this study, fractional factorial design was used to identify the significant factors having the largest effects on the forward extraction process of jacalin using reverse micelles formed by an anionic surfactant, sodium bis(2-ethylhexyl) sulfosuccinate (AOT). Effect of five different factors, namely the aqueous phase pH, surfactant (AOT) concentration, salt (NaCl) concentration, contact time and volume ratio of organic to aqueous phase on the forward extraction efficiency were investigated using the 25-1 fractional factorial design. The results showed that only the aqueous phase pH, NaCl concentration and concentration of AOT had significant effects on the forward extraction efficiency of jacalin into AOT reverse micelles. Results obtained indicated that the fractional factorial design was a useful method for screening many process factors with minimum number of experiments.

Role of Surfactants Cetyl Pyridinium Chloride (CPC) and Cetyltrimethyl Ammonium Bromide (CTAB) in the Reverse Micellar Extraction of Ternary Mixture of Acid Dyes from Textile Effluent

AbstractTextile industry produces colored effluent which is the major cause of water pollution. Removal of color from effluent is very important before discharging. Extraction of ternary mixture of anionic dyes, namely, Acid Orange 7 (AO-7), Acid Blue 1(AB-1) and Acid Yellow 17 (AY-17) was carried out from aqueous solutions using reverse micelles of Cetyl Pyridinium Chloride (CPC) and Cetyltrimethyl ammonium bromide (CTAB) in isoamyl alcohol. Extraction efficiency of CPC and CTAB was studied under various experimental conditions, such as pH of feed solution, concentration of surfactant, extraction time, temperature and initial dye concentration. Under the optimized experimental conditions, 93–98 % of dyes were extracted. It was observed that CTAB is more efficient than CPC for extraction of dyes from aqueous medium.

Construction of Surfactin--Polysorbate-80 reversed micelle systems and its application on extraction of cellulasee

This work studied a novel method to construct a mixed reverse micelle system and its application in extracting and purifying enzymes. Mixed reversed micellar extraction formed by biosurfactant surfactin and ionic surfactant polysorbate-80 was applied for extracting cellulase. 40% surfactin and 60% polysorbate-80 were found to be the best system. The activity and protein recovery were obtained with the best conditions: in forward extraction, surfactant concentration was 3.0 mM; pH 7.0; temperature 40°C; KCl 0.05 M. In backward extraction, pH 7.5; temperature 40°C; KCl 0.1 M.

Optimization and in Silico Analysis of a Cold-Adapted Lipase from an Antarctic Pseudomonas sp. Strain AMS8 Reaction in Triton X-100 Reverse Micelles

A moderate yield of a purified enzyme can be achieved by using the simple technique of reverse micellar extraction (RME). RME is a liquid–liquid extraction method that uses a surfactant and an organic solvent to extract biomolecules. Instead of traditional chromatographic purification methods, which are tedious and expensive, RME using the nonionic surfactant Triton X-100 and toluene is used as an alternative purification technique to purify a recombinant cold-adapted lipase, AMS8. Various process parameters were optimized to maximize the activity recovery of the AMS8 lipase. The optimal conditions were found to be 50 mM sodium phosphate buffer, pH 7, 0.125 M NaCl, and 0.07 M Triton X-100 in toluene at 10 °C. Approximately 56% of the lipase activity was successfully recovered. Structural analysis of the lipase in a reverse micelle (RM) was performed using an in silico approach. The predicted model of AMS8 lipase was simulated in the Triton X-100/toluene reverse micelles from 5 to 40 °C. The lid 2 was slightly opened at 10 °C. However, the secondary structure of AMS8 was most affected in the non-catalytic domain compared to the catalytic domain, with an increased coil conformation. These results suggest that an AMS8 lipase can be extracted using Triton X-100/water/toluene micelles at low temperature. This RME approach will be an important tool for the downstream processing of recombinant cold-adapted lipases.

Green and Efficient Reverse Micellar Extraction and Recovery of Mixed Ionic Dyes from Textile Effluent

Abstract Colored effluents discharged from textile and other industries pose a major threat to the environment. These effluents must be decolorized before discharging into the main streams. The present study explores a solvent extraction method for selective removal of dye from a cationic and anionic dye mixture by treating the dye mixture with reverse micelles of anionic and cationic surfactants in a stepwise manner. The effect of different parameters such as surfactant and dye concentration was also investigated. It has been found that the percentage extraction of dyes increased with increasing surfactant concentration and decreasing dye concentration. UV-visible absorption spectra were analyzed to evaluate the state of the dyes in bulk water before and after extraction with reverse micelles. The dyes were recovered successively by backward extraction and then reused. Their K/S values were also measured.

Separation of Monascus pigments from extractive fermentation broth with a high concentration of triton X-100

ABSTRACTIn this study, Monascus pigments were separated from the extractive fermentation broth by the combination of reverse micellar extraction and macroporous resins (MACR) adsorption with the alkalizing S-8 MACR (S-8 ALR). A 95.07% pigment adsorption was achieved together with a relative low Triton X-100 co-adsorption that was only 2.53%. The liquid intraparticle diffusion and film diffusion limit the pigment adsorption rate. A 79.48–84.03% of the pigment could be eluted from the resin using the methanol solution containing a 0.5% (w/v) hydrochloric acid. The S-8 ALR could be recovered for cycle use without a deficit of the adsorption or desorption efficiency.Abbreviations: ACR—Acidified resin; ALR—Alkalizing resin; HPLC—High performance liquid chromatography; PSM—pigment- surfactant micelle; TEM—Transmission electron microscopy.

Extraction and purification of polyphenol oxidase: A review

Abstract Polyphenol oxidase (PPO) is a copper containing enzyme widely occurred in many plants, animals, bacteria and some fungi. Polyphenol oxidase has wide applications in the field of biosensor formation, food industry, medicine and waste water treatment. Extraction and purification of PPO was studied by many researchers from wide variety of plants and fermentation process using various methods such as salt precipitation, three phase partitioning (TPP), aqueous two phase extraction (ATPE), reverse micellar extraction (RME) and chromatographic techniques. Physio-chemical characterization of the same was performed by the researchers using different techniques. Diversity in the resources of the enzyme also made scientist to study its substrate specificity, optimized activity conditions and inhibitors in detail. Aim of this review is to summarize the detail study regarding extraction and purification of polyphenol oxidase performed by researchers from wide variety of sources.

A Green and Efficient Method for the Preconcentration and Determination of Gallic Acid, Bergenin, Quercitrin, and Embelin from Ardisia japonica Using Nononic Surfactant Genapol X-080 as the Extraction Solvent.

A simple cloud point preconcentration method was developed and validated for the determination of gallic acid, bergenin, quercitrin, and embelin in Ardisia japonica by high-performance liquid chromatography (HPLC) using ultrasonic assisted micellar extraction. Nonionic surfactant Genapol X-080 was selected as the extraction solvent. The effects of various experimental conditions such as the type and concentration of surfactant and salt, temperature, and solution pH on the extraction of these components were studied to optimize the conditions of Ardisia japonica. The solution was incubated in a thermostatic water bath at 60°C for 10 min, and 35% NaH2PO4 (w/v) was added to the solution to promote the phase separation and increase the preconcentration factor. The intraday and interday precision (RSD) were both below 5.0% and the limits of detection (LOD) for the analytes were between 10 and 20 ng·mL−1. The proposed method provides a simple, efficient, and organic solvent-free method to analyze gallic acid, bergenin, quercitrin, and embelin for the quality control of Ardisia japonica.

Purification of recombinant human interferon gamma from fermentation broth using reverse micellar extraction: A process optimization study

ABSTRACTIn the present investigation, we have shown a single step purification of IFN-γ from the fermentation broth of recombinant Kluyveromyces lactis (K. lactis) strain, using cationic surfactant based reverse micellar extraction (RME) system. The forward and back extraction processes were optimized in order to improve the overall extraction process. 78%, 93% and 98% forward extraction efficiencies (FEE) were obtained after the process by using aqueous phase pH 12, 150 mM CTAB and 0.2 M NaCl respectively. Afterwards, back extraction efficiency (BEE) was maximized by the optimization of stripping phase pH, isopropyl alcohol (IPA) and potassium chloride (KCl) concentration by employing Taguchi’s method of orthogonal array. Stripping phase pH 7, 15% IPA & 0.8 M KCl were found to be the best possible parameters producing 83% BEE. The usage of IPA has proven to reduce the effect of micellar-micellar based interaction, which was reflected as an improvement in BEE. This study demonstrates that the reverse micellar system is a highly promising and efficient tool for the direct extraction of recombinant proteins from fermentation broth. Moreover, this system holds a high potential to be an integral part of the downstream process used in biopharmaceutical industry.

Efficient Micellar Extraction of Ternary Mixture of Cationic Dyes from Aqueous Medium

In the present work an attempt has been made to extract ternary mixture of Basic Yellow-2 (BY-2), Basic Red-12 (BR-12) and Basic Blue-26 (BB-26) dyes from simulated textile effluent by solvent extraction using anionic surfactant Sodium Dodecylsulfate (SDS). This study proposes an efficient, fast, simple and effective microemulsion system for the removal of cationic dyes using reverse micelles of anionic surfactant from wastewaters. In this method, dyes are dissolved in aqueous phase and mixed with organic solvent in the presence of surfactant. The effects of the concentration of feed mixture (dye and surfactant), temperature, pH and salt concentration on extraction of dyes from aqueous phase have been studied after the complete separation of two phases. The method poses no environmental threat, so it is an eco-friendly way to treat dye house effluents.