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QUICKSTART GUIDE TO SPE
Appendix 1. Generic SPE methodology for extraction of drugs from biological fluids
a.i EVOLUTE ABN
EVOLUTE ABN is designed for the simultaneous extraction of acidic, basic and neutral drugs from biological fluid
samples. The method below is optimized for the extraction of 100 µL sample volumes, using the 25 mg sorbent
mass.
1. Sample Pre-treatment: Dilute sample (typically plasma) 1:3 (v/v) with aqueous formic acid
(1%, v/v). Add internal standard and mix thoroughly.
2. Conditioning: Condition each column or well with methanol (1 mL).
3. Equilibration: Equilibrate each column or well with aqueous formic acid (0.1%,
v/v, 1 mL).
4. Sample Load: Load sample (typically 400 µL diluted plasma).
5. Interference Wash: Elute interferences with water/methanol (95:5, v/v, 1 mL).
6. Analyte Elution: Elute analytes with methanol (500 µL).
7. Post-extraction: If required, evaporate extracts to dryness and reconstitute in mobile
phase or other suitable solvent prior to analysis.
Request Chemistry Data Sheet TN131 for further details.
a.ii EVOLUTE CX
EVOLUTE CX is designed to efficiently extract basic drugs from biological fluids using a dual retention mechanism.
This allows additional interference elution steps to be used, giving extremely clean extracts free from
proteins, salts and phospholipids, with minimal matrix effects in LC-MS/MS analysis.
The method below is optimised for the extraction of 100 µL sample volumes using the 25 mg sorbent mass
columns or 96-well plates.
1. Sample Pre-treatment: Dilute sample (typically plasma) with ammonium acetate (0.05M, pH 6)
(1:3, v/v). *Alternatively, dilute sample with aqueous formic acid (1%,
1:3, v/v).
2. Conditioning: Condition each column or well with methanol (1 mL).
3. Equilibration: Equilibrate each column or well with ammonium acetate (0.05M, pH6,
1 mL).
4. Sample Load: Load sample (typically 400µL diluted plasma).
5. Interference Wash 1: Rinse each well or column with ammonium acetate (0.05M, pH6, 1 mL).
6. Interference Wash 2: Rinse each well or column with methanol (1 mL).
7. Analyte elution: Elute analytes with methanol: ammonium hydroxide (95:5, v/v,
500 µL).
8. Post extraction: If required, evaporate extracts to dryness and reconstitute in mobile
phase or other suitable solvent prior to analysis.
*Alternative sample pre-treatment is useful for analytes which are strongly protein bound.
Request Chemistry Data Sheet TN139 for further details
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