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Content
Liquid / liquid Extraction I
Liquid / liquid Extraction II
Drawbacks of Liquid/liquid Extraction
Emulsions
Support-Assisted Liquid / liquid Extraction I
Support-Assisted Liquid / liquid Extraction II
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If the analytes are not yet adequately pure, a second extraction of the organic solvent
using an aqueous phase is sometimes used (referred to as a back-extraction). This
approach works best with basic compounds, which can be neutralized (making them more
organic-soluble) by raising the pH of the original sample, extracting into an organic
solvent, then back-extracting into an aqueous phase using an acidic buffer (ionizing the
analytes, thus making them more water-soluble).
Another approach to liquid/liquid extraction for highly water-soluble analytes is extraction
of non-polar, organic-soluble interferences from the sample, leaving the analytes behind in
the aqueous phase. This approach has the specific drawback that the low-volatility
aqueous phase is not readily concentrated, as is an organic solvent.
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Another drawback of liquid/liquid extraction for large numbers of samples is that it is not
well-suited to automation. Other approaches, SPE in particular, are much easier to
automate.
Emulsions
One major drawback of liquid/liquid extraction is the formation of emulsions. Emulsions
occur when the sample contains a high level of surfactant-like compounds (notably
phospholipids), that prevent clean separation of the two phases. With emulsions, a midzone between the two phases is created, having intermediate solubility in each of the two
phases, and making it difficult to quantitatively collect one phase or another.
Emulsion formation
Emulsions often occur with samples where the animal (or human) diet is high in fats.
Thus, emulsions sometimes appear when passing from pre-clinical trials, with animals on
low-fat, controlled diets, to clinical trials, with humans who may be on high-fat diets. This
characteristic problem makes liquid/liquid extraction a less dependable procedure if it is
expected that the same extraction protocol will be used for both pre-clinical and clinical
samples. If this problem is anticipated, it is worth trying high-fat samples during method
development in addition to the standard test matrices.
Emulsions may sometimes be disrupted, or broken-up, by the addition of salt to the
emulsion. The salt changes the capacity of the aqueous phase to accommodate the
marginally-soluble components in the system, thus driving them into the organic phase.
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The organic solvents most commonly used are the same solvents used in conventional
liquid/liquid extraction; ethyl acetate, methyl tert-butyl ether (MTBE), methylene chloride,
hexane, and mixtures thereof.
Another advantage of SALL is that the nature of the column technique allows for superior
automation as compared to conventional liquid/liquid extraction.
Finally, the SALL approach is essentially free of the emulsion problems common to
conventional liquid/liquid extraction. In pharmaceutical applications, in particular, this
allows for the use of the technique across the range of samples from pre-clinical to
clinical, with reduced concern for method ruggedness.
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