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SPE - What can you learn in 5min?

Solid phase extraction (SPE) is a routinely used technique to clean and / or concentrate samples prior to chromatography.

This is how it works :

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An SPE method follows 5 steps :

  1. Sample pretreatment – e.g. pH adjustment, protein precipitation, filtration
  2. Column conditioning – the column is made ready for the sample being loaded
  3. Sample loading – the sample is loaded onto the cartridge
  4. Column wash – interferences are washed from the column
  5. Sample elution – the sample is eluted by a suitable solvent and collected – interference left behind on cartridge

The three main parameters to optimize are the choice of column sorbent, the choice of wash solvent and the choice of eluting solvent.


One of main problems encountered with an SPE method is poor or non-repeatable recovery of the sample.
The three main reasons for this are (using mechanism 1 as an example) :

1) Incorrect choice of sorbent bed. The sorbent bed must FULLY retain the analyte of interest, any leaching of sample during load or wash (known as breakthrough) will cause poor recoveries.
There is a diverse range of stationary phases available – reversed phase, ion exchange, mixed mode (blend of reversed phase and ion exchange on the same column) and normal phase. Some knowledge of the chemical properties of the analyte(s) of interest is required in order to produce an effective method.


2) Incorrect wash solvent. The wash solvent must be strong enough to remove the most interference without eluting any sample.

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3) Incorrect elution solvent. The eluting solvent must be strong enough to elute 100% of the sample, but weak enough to leave as much interfering contaminants on the column as possible.

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Finding the correct balance of these three parameters is critical in achieving a reliable and robust method and can be intimidating to new users. However, the choice of solvents is strongly dictated by the stationary phase choice.

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