ESI-MS/MS

Warfarin in Plasma.
Experiment procedures
1. Sample treatment
SPE (solid phase extraction)
3 layer polimers used to extract hydrophobic compounds. To ''warm up'' the polymer, 5 mL of MeOH is added to SP (Oasis HLB, 200mg, water corporation, Milford, MA, US). use vacumn pump to make MeOH pass through SP.
Then use 5mL MiliQ water pouring on the SP.
After that add 0.5 mL filtered through 0.45 uL membrane to the SP.
water+5% MeOH to wash SPE column
Elute SPE with 5mL MeOH followed by 3 mL ethyl acetate.
collect elute then evaporate to 20uL
- dilute in up to 1mL in mobile phase (10% acetonitrile in water(0.1% formic acid)
Run LC-MS/MS
before that prepare 3 standard 1000,800,600 ppb warfarin.