See Patented via Nth Cycle for metal electro-extraction process. [1]
Electroextraction (EE) is a sample enrichment technique that focuses charged analytes from a large volume of one phase into a small volume of aqueous phase through the application of an electric current. [2] The technique was originally developed as a separation technique for chemical engineering, but has since been coupled to capillary electrophoresis and liquid chromatography–mass spectrometry as a means of improving limits of detection, analysis time, and selectivity. [2] [3] [4] The use of EE-CE has made capillary electrophoresis more applicable to use in the pharmaceutical industry.
The term electroextraction is used to describe multiple processes. In this article, electroextraction describes the extraction of charged particles through a liquid phase barrier. The term can be used to describe electrowinning, which is the extraction of metals from their compounds via electrochemical processes. Electroextraction also describes a process of permeating bioproducts from a cell membrane using an electric field. [5]
Johann Stichlmair developed electroextraction at the University of Essen in Germany in 1987 as an improvement on liquid-liquid extraction in an electric field. Electric fields are applied to enhance the demixing of a sample in a two-phase system. However, as current flows through the system, the resulting convective mixing disrupts separation. Electroextraction corrects for this. Two or three liquid phases that are electrically conductive and immiscible with one another are kept between electrodes, and upon addition of an electric field, charged particles travel from one phase to another separating anions and cations. A two-phase system brings anions into one phase and cations into the other. A three phase system extracts anions and cations into the two outer phases while leaving uncharged particles in the middle phase. [6] Convective mixing is restricted to each phase and does not travel between phases. A diagram is given in figure 1. Organic phases that are used typically have small amounts of water added in order to be conductive. Other possible phases include mixtures of water and highly polymerized substances, or water with non-ionic surfactants. [3] Electroextraction has also influenced the development of similar electrophoretic separation techniques involving a membrane between the two-phase systems.
Electroextraction has been successfully employed in the separation of dyestuffs from wastewater. Electroextraction is better suited over other techniques for its ability to extract small amounts of dye from very dilute solutions. EE has also been effectively used in the separation of amino acids. This separation was done using an aqueous two-phase system of dextran-polyethylene glycol-water to stabilize the amino acids. [7] The velocity of a particle crossing the phase barrier is directly proportional to the strength of the applied electric field, so 100% separation is achieved with a strong enough field. [8]
A diagram of an electroextraction apparatus is shown in figure 2. The apparatus consists of a vial with a conical bottom, a grounded platinum electrode, a capillary to inject the aqueous solution, and an adjustable gold anode with a circular bottom that contacts the entire organic phase.
EE is also often performed in a capillary electrophoresis capillary. This is referred to as capillary electroextraction, or cEE. In this set up, shown in figure 3, a capillary containing aqueous phase is placed in a vial of organic phase and surrounded by a hallow cathode. The outlet of the capillary is then grounded. [9]
When EE is coupled to isotachophoresis coupled to capillary electrophoresis, limits of detection decrease to the nanomolar range and isotachophoresis takes only a few minutes to complete. Similar limits of detections are obtained when EE is coupled to liquid chromatography-mass spectrometry. [2] Similar limits of detections are obtained when EE is coupled to liquid chromatography-mass spectrometry. [4]
EE is ideal for pharmaceuticals with low concentrations of active ingredient, such as those containing proteins and peptides, because of its ability to lower limits of detection for liquid chromatography and capillary electrophoresis. [10] In addition, sample enrichment by EE helps overcome the low injection volumes and short optical path length of UV-Vis detectors that accompany CE. [2] EE coupled to CE has been used to separate and analyze antisense oligonucleotides. Antisense oligonucleotides inhibit protein expression from their complementary base pair sequence and can treat certain diseases and genetic disorders. [11] EE coupled to liquid chromatography also successfully detects low concentration metabolites in urine for the purpose of studying metabolic processes. [12] In addition, EE-ITP-CE has been used in the determination of the drugs clenbuterol, salbutamol, terbutaline, and fenoterol. [13]
Analytical chemistry studies and uses instruments and methods to separate, identify, and quantify matter. In practice, separation, identification or quantification may constitute the entire analysis or be combined with another method. Separation isolates analytes. Qualitative analysis identifies analytes, while quantitative analysis determines the numerical amount or concentration.
In chemical analysis, chromatography is a laboratory technique for the separation of a mixture into its components. The mixture is dissolved in a fluid solvent called the mobile phase, which carries it through a system on which a material called the stationary phase is fixed. Because the different constituents of the mixture tend to have different affinities for the stationary phase and are retained for different lengths of time depending on their interactions with its surface sites, the constituents travel at different apparent velocities in the mobile fluid, causing them to separate. The separation is based on the differential partitioning between the mobile and the stationary phases. Subtle differences in a compound's partition coefficient result in differential retention on the stationary phase and thus affect the separation.
High-performance liquid chromatography (HPLC), formerly referred to as high-pressure liquid chromatography, is a technique in analytical chemistry used to separate, identify, and quantify each component in a mixture. It relies on pumps to pass a pressurized liquid solvent containing the sample mixture through a column filled with a solid adsorbent material. Each component in the sample interacts slightly differently with the adsorbent material, causing different flow rates for the different components and leading to the separation of the components as they flow out of the column.
Micellar electrokinetic chromatography (MEKC) is a chromatography technique used in analytical chemistry. It is a modification of capillary electrophoresis (CE), extending its functionality to neutral analytes, where the samples are separated by differential partitioning between micelles and a surrounding aqueous buffer solution.
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Liquid chromatography–mass spectrometry (LC–MS) is an analytical chemistry technique that combines the physical separation capabilities of liquid chromatography with the mass analysis capabilities of mass spectrometry (MS). Coupled chromatography - MS systems are popular in chemical analysis because the individual capabilities of each technique are enhanced synergistically. While liquid chromatography separates mixtures with multiple components, mass spectrometry provides spectral information that may help to identify each separated component. MS is not only sensitive, but provides selective detection, relieving the need for complete chromatographic separation. LC-MS is also appropriate for metabolomics because of its good coverage of a wide range of chemicals. This tandem technique can be used to analyze biochemical, organic, and inorganic compounds commonly found in complex samples of environmental and biological origin. Therefore, LC-MS may be applied in a wide range of sectors including biotechnology, environment monitoring, food processing, and pharmaceutical, agrochemical, and cosmetic industries. Since the early 2000s, LC-MS has also begun to be used in clinical applications.
Electrochromatography is a chemical separation technique in analytical chemistry, biochemistry and molecular biology used to resolve and separate mostly large biomolecules such as proteins. It is a combination of size exclusion chromatography and gel electrophoresis. These separation mechanisms operate essentially in superposition along the length of a gel filtration column to which an axial electric field gradient has been added. The molecules are separated by size due to the gel filtration mechanism and by electrophoretic mobility due to the gel electrophoresis mechanism. Additionally there are secondary chromatographic solute retention mechanisms.
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Two-dimensional chromatography is a type of chromatographic technique in which the injected sample is separated by passing through two different separation stages. Two different chromatographic columns are connected in sequence, and the effluent from the first system is transferred onto the second column. Typically the second column has a different separation mechanism, so that bands that are poorly resolved from the first column may be completely separated in the second column. Alternately, the two columns might run at different temperatures. During the second stage of separation the rate at which the separation occurs must be faster than the first stage, since there is still only a single detector. The plane surface is amenable to sequential development in two directions using two different solvents.
Capillary electrophoresis–mass spectrometry (CE-MS) is an analytical chemistry technique formed by the combination of the liquid separation process of capillary electrophoresis with mass spectrometry. CE-MS combines advantages of both CE and MS to provide high separation efficiency and molecular mass information in a single analysis. It has high resolving power and sensitivity, requires minimal volume and can analyze at high speed. Ions are typically formed by electrospray ionization, but they can also be formed by matrix-assisted laser desorption/ionization or other ionization techniques. It has applications in basic research in proteomics and quantitative analysis of biomolecules as well as in clinical medicine. Since its introduction in 1987, new developments and applications have made CE-MS a powerful separation and identification technique. Use of CE-MS has increased for protein and peptides analysis and other biomolecules. However, the development of online CE-MS is not without challenges. Understanding of CE, the interface setup, ionization technique and mass detection system is important to tackle problems while coupling capillary electrophoresis to mass spectrometry.
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