Theory of SPE
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Theory of SPE
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SPE
Nonequilibrium, exhaustive removal of chemical constituents from a flowing liquid sample via retention on a contained solid sorbent and subsequent recovery of selected constituents by elution from the sorbent
M.J.M. Wells, Essential guides to method development in solid-phase extraction, in I.D. Wilson, E.R. Adlard, M. Cooke, and C.F. Poole, eds., Encyclopedia of Separation Science, Vol. 10, Academic Press, London, 2000, pp. 4636-4643.
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Ref.: E.M. Thurman, and M.S. Mills, “Solid-Phase Extraction
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Four Typical Steps of SPE
Conditioning
Loading Adsoption
Washing
Elution
Loading/Adsorption
Gentle vacuum, or pump
At reasonable rate, depend on column dimension, particle size
Small particles, more efficient, permit faster flow rate
The sorbent should not be allow to go dry at any point
Air in the column prevent efficient interfacial contact
between liquid and solid phase
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Washing
Remove interferences coadsorbed from the SPE column
The wash solution must not be too strong to partially eluted the analyte of interest
For RP-SPE, when the eluting solvent is immiscible with water the sorbent must be dried so that the eluting solvent can interact with all area of the sorbent
Gentle vacuum for a few minutes
Compressed air or N2
Centrifugation
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Elution
Eluting solvent should be strong enough to completely removed adsorbed analytes from the sorbent as small a volume as possible (small k) Compatible with the analytical method (i.e., Low BP –GC)
Free from impurity
Low cost and non toxic
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Choice of Eluting Solvent
Reversed Phase SPE Nonpolar solvent
Methanol
Acetonitrile
Ethyl acetate
Acetone
Methylene chloride Normal Phase SPE
A function of the
eluotropic strength
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Eluting Solvent
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Parameters Affecting N
Flow rate
Particle size and plate number
Column length
Temperature
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Sample Breakthrough
The analytes are no longer retained by the sorbent.
The capacity of the sorbent has been exceeded or overloaded
The maximum sample volume that can be passed through the SPE column
Retention volume (V R) and the number of theoretical plates (N)
Frontal Chromatography
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k w = Capacity factor of solute eluted by water obtained by
linear extrapolation from methanol -water solutions
k w can be estimated from a linear relationship between the average log k w and log P oct
Estimation of Breakthrough Volume
V R = V o (1+k w )
Log k w = 0.988 (±0.051) log P oct + 0.02 (±0.060)P oct = Octanol -water partition coefficient
(The affinity of the solute for the stationary phase)Ref.: T. Braumann , J. Chromatogr ., 373, 191-225 (1986)
Recovery
Recovery = sorption efficiency x desorption efficiency
Sample pH
Sample volume
Sorbent mass
Sample concentration
Eluting Strength
Solvent volume
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SPE Method Development
What is solute structure? The clue to effective SPE
Identify the goal
Obtain physical constants
Choose the mode (mechanism) of SPE
Elute the SPE cartridge
Perform the sorption experiment and determine breakthrough
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Troubleshooting
Incomplete elution
Increase eluting solvent, change solvent, pH adjustment, back elution
Breakthrough of analyte
Increase the amount of sorbent, change the sorbent, reduce
flow rate
Interfering substances
Wash with solvent selectively elute the interferences, clean up the eluent with another sorbent
SPE bleed, wash the sorbent, change the solvent
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