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SiliaPrep Tips Micro-SPE Cartridges Experimental Procedure

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The following lines present the general experimental procedure for the purification and enrichment of small molecules, peptides and proteins using SiliaPrep Tips Micro-SPE Cartridges.

1. Conditionning Step
Attach the SiliaPrep Tips to a micropipette.
Aspirate / expel the elution solution 5 times and the binding solution 3 times.

2. Loading Step
Aspirate / expel the sample 20 to 50 times to allow compounds to adsorb onto the sorbent.

3. Washing Step
Aspirate / expel the binding solution 10 times and discard the expelled solution each time.

4. Elution Step
Aspirate / expel the elution solution 10 times and collect the expelled solution in a suitable clean tube. Repeat with a fresh portion of elution solution if you want to be sure to collect all of the adsorbed compounds.
Note: repeat 3 - 5 times for carbon black sorbent.

 

SiliaPrep Tips Micro-SPE Cartridges Experimental Procedure - Loading Step SiliaPrep Tips Micro-SPE Cartridges Experimental Procedure - Washing Step SiliaPrep Tips Micro-SPE Cartridges Experimental Procedure - Elution Step

 

SiliaPrep Tips Micro-SPE Application

Micro-Extraction of Dextromethorphan from Plasma
Cartridge
SiliaPrepX Tips C18 10 μl/30 μg
Part Number: SPET-C18-T1
Sample Pretreatment
8 μl of plasma sample was mixed with 2 μl of internal standard (Dextromethorphan-d3 at 10 ng/ml in Methanol)
Conditionning step
8 μl of Methanol (10 aspirate / expel)
Equilibration step
8 μl of Water (10 aspirate / expel)
Loading step
Plasma sample (30 aspirate / expel)
Washing step
8 μl of water (10 aspirate / expel) then 8 μl of 25 % Methanol in water (10 aspirate / expel)
Elution step
8 μl of Acetonitrile (30 aspirate / expel)
Further treatment
Quantification by LDTD/MS/MS (collaboration with Phytronix)
Recovery at 10 ng/ml
- Dextromethorphan: 86 %
- Dextromethorphan d-3: 80 %

 

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