Materials and Reagents
Reagent
1. Ammonium Acetate
2. Methanol
2. Methoyl Tertiary Butyl Ether(MTBE)
Material
1. EP tube
Procedure
Step 1
TissueMince tissue into small pieces in 0.1% ammonium acetate water solution followed by homogenization.
Serum/Plasma
Step 2
100 μL Step 1 sample aliquot add 750 μL methanol and vortex for at least 10s in the EP tube.
Add 2.5 mL MTBE and mix well by vortex or ultrasonic
Incubate the mixture at RT for 1 h in a shaker.
Step 3
Add 625 μL H2O and mix well.
Stay at RT for 10 min.
Centrifuge at 1000 g for 10 min.
Step 4
Take the supernate (organic phase) to another EP tube.
Re-extract the lower phase with 2 mL of extraction agent (will it be incubate as Step 2?).
Combine the 4.2 supernate with 4.1.
Step 5
Concentrate the combined supernate with nitrogen or vacuum centrifuge.
Redissolve the 5.1 by 100 μL mobile phase A (ACN:H2O, 60%:40%) for injection.
Recipes