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Non-polar SPE for extraction of drugs from biological fluids

Non-polar SPE for extraction of drugs from biological fluids. Argonaut Technologies 1101 Chess Drive, Foster City CA 94404 USA. Relative selectivity of various sample preparation techniques. Protein precipitation Liquid-liquid extraction Non-selective resin SPE (hydrophobic only)

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Non-polar SPE for extraction of drugs from biological fluids

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  1. Non-polar SPE for extraction of drugs from biological fluids Argonaut Technologies 1101 Chess Drive, Foster City CA 94404 USA

  2. Relative selectivity of various sample preparation techniques • Protein precipitation • Liquid-liquid extraction • Non-selective resin SPE(hydrophobic only) • Supported liquid-liquid extraction • C18 silica based SPE • C2 silica based SPE • Mixed-mode SPE - resin based sorbents • Ion exchange SPE - silica based sorbents • Mixed-mode SPE - silica based sorbents • Immunoaffinity, MIPs Non selective Dirty extracts Highly selective Clean extracts

  3. Typical non-polar SPE procedure 1. Sample pre-treatment 2. Column solvation 3. Column equilibration 4. Sample application 5. Interference elution 6. Analyte elution

  4. Non-polar SPE 1 2/3 4 5 6

  5. Generic non-polar SPE method Column: ISOLUTE XXX 25mg / 1ml or 96-well plate Adjust pH 1 mL methanol 500L 0.05Mbuffer Apply sample (0.1 – 1 mL) 500 L buffer / MeOH 95:5 500 L methanol Sample pre-treatment Column solvation Column equilibration Sample application Interference elution Analyte elution

  6. Sorbent screening - breakthrough C8 C18 C2 CN PH

  7. Sorbent Screening – analyte elution C18 C8 PH

  8. Increase conc. MeOH to improve extract cleanliness (check for analyte breakthrough) Optimising the method Acidify to maximise recovery of acidic analytes; add base to maximise recovery of basic analytes Adjust pH Methanol 0.05Mbuffer Apply sample Buffer / MeOH 95:5 Methanol Ensures good flow in 96-well SPE + maximises capacity Same pH as sample Optimise for minimum elution volume, including soak step

  9. Limitations with non-selective sample preparation • Potential interferences are not efficiently removed from the sample, leading to: • Matrix effects • Inaccurate recoveries • Variable LC/MS quantification • Shorter HPLC column life

  10. Summary • Non selective nature of the non-polar sorbents allows a wide range of drug candidates to be extracted • Parallel screening rapidly identifies optimum sorbent • Narrow particle size distribution allows sorbent mass optimisation • Optimum sorbent mass leads to minimum elution volumes

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