Solid Phase Extraction (SPE)
We offer SPE columns, cartridges, multi 96‑well plates and accessories (e.g., vacuum manifolds) for your SPE applications. Beside standard silica gel based SPE phases our broad product range includes special phases and polymeric materials like CHROMABOND HR‑X or HLB as well as QuEChERS mixes or automated SPE products.
SPE is a powerful method for sample preparation and is used by most chromatographers today.
About 30 years ago MACHEREY‑NAGEL designed and introduced CHROMABOND SPE cartridges containing silica-based adsorbents.
SPE is a form of digital (step-wise) chromatography designed to extract, partition, and / or adsorb one or more components from a liquid phase (sample) onto a stationary phase (adsorbent or resin). An adsorbed substance can be removed from the adsorbent by stepwise increase of elution strength of the eluent (step gradient technique).
SPE extends a chromatographic system’s lifetime, improves qualitative and quantitative analysis, and the demand placed on an analytical instrument is considerably lessened.
- Environmental analysis
- Pharmaceutical and biochemical analysis
- Organic chemistry
- Food analysis
Video: Setup of SPE vacuum manifold
Video: Solid phase extraction demonstration
Main steps of the SPE procedure
1) Conditioning Conditioning of the adsorbent is necessary in order to ensure reproducible interaction with the analyte. Conditioning, also called solvation, results in a wetting of the adsorbent and thus produces an environment, which is suitable for adsorption of the analyte. Nonpolar adsorbents are usually conditioned with 2–3 column volumes of a solvent, which is miscible with water (methanol, THF, 2-propanol etc.), followed by the solvent in which the analyte is dissolved (pure matrix, e.g., water, buffer). Polar adsorbents are conditioned with nonpolar solvents. After the conditioning step the adsorbent bed must not run dry, because otherwise solvation is destroyed (deconditioning).
2) Sample application (adsorption) Sample application can be performed with positive or negative pressure with a flow rate of ~3 mL/min. Sample volumes vary from a few mL up to liters.
3) Washing Washing of the adsorbent is usually achieved with a special wash solution; however, in some cases it may not be necessary. If the polarity difference between wash solution and eluent is very large, or if both are not miscible, drying of the adsorbent bed after washing is recommended to improve elution and recovery.
4) Elution Elution with a suitable eluent should not be too fast. The elution speed depends on the column or cartridge dimension and the quantity of adsorbent (about 1 mL/min).
Since analytes can either be adsorbed on the SPE packing material or directly flown through while the interfering substances are retained, two general separation procedures are possible – both cases are shown in the figure below.
a) Retention of the analyte (top path)
- Analyte molecules are enriched on the adsorbent
- Interfering components and solvent molecules (matrix) are not retained
- Remaining interfering components are washed from the adsorbent
- The analyte is removed from the adsorbent by elution with a suitable solvent
b) Retention of interfering components (bottom path)
- Analyte molecules show no interaction with the adsorbent
- Interfering components and solvent molecules (matrix) are retained
- Analyte molecules are “washed” from the adsorbent
- The solid phase is used to “filter” the sample
- We have developed the widest range of phases for SPE based on silica, polymeric and other materials.
- CHROMABOND columns, cartridges and 96-well plates are manufactured from polypropylene (PP) with lowest content of extractables (plasticizers, stabilizers, …) offering blank value free results when using most common solvents.
- CHROMABOND columns are also available as glass columns (3 mL and 6 mL volume).
- The high quality CHROMABOND adsorbents are kept in place by chemically very inert polyethylene (PE) or glass fiber (GF) filter elements.