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Solid phase extraction
Solid-phase extraction (SPE) is a separation process that is used to extract compounds (called analytes) from a mixture of impurities. Analytical laboratories use solid phase extraction to concentrate and purify samples for analysis. Solid phase extraction can be used to isolate analytes of interest from a wide variety of matrices, including urine, blood, water samples, beverages, soil, animal tissue, and consumer products.
The separation ability of solid phase extraction is based on the preferential affinity of desired or undesired solutes in a liquid, mobile phase for a solid, stationary phase through which the sample is passed. Impurities in the sample are either washed away while the analyte of interest is retained on the stationary phase, or vice-versa. Analytes that are retained on the stationary phase can then be eluted from the solid phase extraction cartridge with the appropriate solvent.
The stationary phase is usually packed in a syringe-shaped cartridge or a 96 well plate, which can be mounted on a commercially available extraction manifold. A typical SPE manifold can accommodate 12 or 24 SPE cartridges so that many samples can be processed simultaneously. Many SPE manifolds are equipped with a vacuum port; application of a vacuum speeds up the extraction process by pulling the liquid sample through the stationary phase. The analyte is collected with a sample tube rack inside the manifold, which collects the sample as it is eluted off the cartridge.
Solid phase extraction cartridges are available with a variety of different stationary phases, each of which can separate analytes by a different chemical mechanism. Most stationary phases are based on a bonded silica material that is derivatized with a specific functional group. Some of these functional groups include hydrocarbon chains of variable length (for reversed phase SPE), quaternary ammonium or amino groups (for anion exchange), and sulfonic acid or carboxyl groups (for cation exchange).
Additional recommended knowledge
A typical solid phase extraction involves four basic steps. First, the cartridge is equilibrated with a non-polar solvent, which wets the surface and penetrates the bonded phase. Then water, or buffer of the same composition as the sample, is typically washed through the column to wet the silica surface. The sample is then added to the cartridge. As the sample passes through the stationary phase, the analytes in the sample will interact and retain on the sorbent while the solvent, salts, and other impurities pass through the cartridge. After the sample is loaded, the cartridge is washed with buffer or solvent to remove further impurities. Then, the analyte is eluted with a non-polar solvent or a buffer of the appropriate pH.
Reversed phase SPE
Reversed phase SPE separates analytes based on their polarity. The stationary phase of a reversed phase SPE cartridge is derivatized with hydrocarbon chains, that retain compounds of mid to low polarity due to the hydrophobic effect. The analyte can be eluted by washing the cartridge with a non-polar solvent, which disrupts the interaction of the analyte and the stationary phase.
Ion exchange SPE
Ion exchange sorbents separate analytes based on electrostatic interactions between the analyte of interest and the positively charged groups on the stationary phase. For ion exchange to occur, both the stationary phase and sample must be at a pH where both are charged.
Anion exchange sorbents are derivatized with positively charged functional groups that interact and retain negatively charged anions, such as acids. Strong anion exchange sorbents contain quaternary ammonium groups that have a permanent positive charge in aqueous solutions, and weak anion exchange sorbents use amine groups which are charged when the pH is below about 9. Strong anion exchange sorbents are useful because any strongly acidic impurities in the sample will bind to the sorbent and usually will not be eluted with the analyte of interest; to recover a strong acid a weak anion exchange cartridge should be used. To elute the analyte from either the strong or weak sorbent, the stationary phase is washed with a solvent that neutralizes the charge of either the analyte, the stationary phase, or both. Once the charge is neutralized, the electrostatic interaction between the analyte and the stationary phase no longer exists and the analyte will elute from the cartridge.
Cation exchange sorbents are derivatized with functional groups that interact and retain positively charged cations, such as bases. Strong cation exchange sorbents contain aliphatic sulfonic acid groups that are always negatively charged in aqueous solution, and weak cation exchange sorbents contain aliphatic carboxylic acids, which are charged when the pH is above about 5. Strong cation exchange sorbents are useful because any strongly basic impurities in the sample will bind to the sorbent and usually will not be eluted with the analyte of interest; to recover a strong base a weak cation exchange cartridge should be used. To elute the analyte from either the strong or weak sorbent, the stationary phase is washed with a solvent that neutralizes ionic interaction between the analyte and the stationary phase.
|This article is licensed under the GNU Free Documentation License. It uses material from the Wikipedia article "Solid_phase_extraction". A list of authors is available in Wikipedia.|