Protein Precipatation vs. Ion Suppression of Medium Polar Drugs on strata Impact Square Well Plate

Application Detail (App ID: 15712)

Part No:00F-4439-Y0
Dimensions:150 x 3 mm ID
Elution Type:Gradient
Elution A:0.1% Formic acid/Water
Elution B:0.08% Formic acid/Acetonitrile
Gradient Profile:
Step No. Time (min) Pct A Pct B
1 0 90 10
2 5 50 50
3 7 50 50

Flow Rate:0.5 mL/min
Col. Temp: ambient
Detection: Mass Spectrometer (MS) @ 249 amu (22°C)
Note:Protein Precipitation Protocol:
Phase: Strata Impact Square Well Plate, 2 mL (CEO-7565)
1. Dispense 300 uL acetonitrile into each well using an automatic pipettor.
2. Place the protein precipitation plate onto a suitable 96-well vacuum manifold . Make sure that a 96-well collection plate is positioned inside the manifold to collect the filtrate.
3. Dispense 100 uL of Porcine plasma into each well ( acetonitrile:plasma = 3:1). Let it stand for 2 mins (no vortex /mixing required).
4. Apply 5-10" of mercury for 30-40 secs.
5. Collect the filtrate and blow down to dryness under slow stream of nitrogen @ 40 deg. C.
6. Reconstitute with 100 uL of mobile phase containing 10.0 ng of analye.
Note: For ion suppression or enhancement estimation, a set of 4 blank (100 uL of water instead of plasma) was run in parallel.

Filtrate looked very clean and clear
Analyte logP m/z % Variation Effect
1. Pindolol 1.75 249 15% Suppression
2. Metoprolol 1.88 268 13% Suppression

Analyte Details

Analyte Detail not available