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SPE method development interactive learning tool

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This month, we look at a simple and structured approach to developing a reversed phase Solid Phase Extraction (SPE) method.

We’ve provided a interactive learning tool to allow you to evaluate different sets of conditions and see what effect they have on the resulting chromatogram.

With this type of SPE, the sample is loaded onto the cartridge and is fully retained by the sorbent bed. While retained to the bed, the sample is washed with a suitable solvent that removes poorly retained interferences, whilst not eluting any of the target analyte. Find out more here >>

We then elute the target analyte using the lowest strength organic solution that removes all the target, whilst leaving behind any strongly retained interferences.

In the interactive experiment below choose a suitable wash and elution percentage that gives the chromatogram with the least interference and a peak height of not less than 10000 for the target (blue - X) peak.

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To perform an approach like this in the lab you would prepare the following solutions :

  1. Condition the cartridge in the normal way

  2. Load the sample onto the cartridge

  3. Wash the sample with an appropriate amount (depending on bed size) of Sol 1

  4. Collect the liquid eluting from the column for analysis of analyte of interest

  5. Repeat steps 3 and 4 sequentially with Sol 2 – 11

  6. Perform an additional 2 washes (3 in total) using sol 11

  7. Plot % organic (0 – 100) on the x-axis and analyte concentration on y-axis

Aqueous Organic Solution Identity
100 0 Sol 1
90 10 Sol 2
80 20 Sol 3
70 30 Sol 4
60 40 Sol 5
50 50 Sol 6
40 60 Sol 7
30 70 Sol 8
20 80 Sol 9
10 90 Sol 10
0 100 Sol 11

The wash solution is the highest Sol solution which gives no analyte concentration.

The elution solution is the lowest Sol solution in which all analyte has eluted (i.e. analyte conc. on y-axis has returned to 0). The graph below illustrates this:





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