Protein Precipatation vs. Ion Suppression of Medium Polar Drugs on strata Impact Square Well Plate
LC Conditions
Column
Gemini 3 µm C18 110 Å, LC Column 150 x 3 mm, Ea
Brand
Gemini
Part No
00F-4439-Y0
Phase Name
C18
Elution Type
Gradient
Mobile Phase
A: 0.1% Formic acid/Water
B: 0.08% Formic acid/Acetonitrile
Gradient Profile
Step No.
Time(min)
%A
%B
1
0
90
10
2
5
50
50
3
7
50
50
Flow Rate
0.50 mL/min
Temperature
22°C
Detection
Mass Spectrometer (MS) @ (22°C)
Detection Info
Sample Notes
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. Observation: Filtrate looked very clean and clear Results: Analyte logP m/z % Variation Effect 1. Pindolol 1.75 249 15% Suppression 2. Metoprolol 1.88 268 13% Suppression
Order Items Used in This Application
- 1. Pindolol
- 2. Metoprolol
-OEChem-06211003152D<br><br> 38 39 0 1 0 0 0 0 0999 V2000<br> 5.4641 -0.5530 0.0000 O 0 0 0 0 0 0 0 0 0 0 0 0<br> 5.4641 1.4470 0.0000 O 0 0 0 0 0 0 0 0 0 0 0 0<br> 3.7320 2.4470 0.0000 N 0 0 0 0 0 0 0 0 0 0 0 0<br> 7.2764 -3.3577 0.0000 N 0 0 0 0 0 0 0 0 0 0 0 0<br> 4.5981 0.9470 0.0000 C 0 0 3 0 0 0 0 0 0 0 0 0<br> 3.7320 1.4470 0.0000 C 0 0 0 0 0 0 0 0 0 0 0 0<br> 2.8660 2.9470 0.0000 C 0 0 0 0 0 0 0 0 0 0 0 0<br> 4.5981 -0.0530 0.0000 C 0 0 0 0 0 0 0 0 0 0 0 0<br> 6.3301 -2.0530 0.0000 C 0 0 0 0 0 0 0 0 0 0 0 0<br> 5.4641 -1.5530 0.0000 C 0 0 0 0 0 0 0 0 0 0 0 0<br> 2.8660 3.9470 0.0000 C 0 0 0 0 0 0 0 0 0 0 0 0<br> 2.0000 2.4470 0.0000 C 0 0 0 0 0 0 0 0 0 0 0 0<br> 6.3301 -3.0530 0.0000 C 0 0 0 0 0 0 0 0 0 0 0 0<br> 7.2764 -1.7482 0.0000 C 0 0 0 0 0 0 0 0 0 0 0 0<br> 4.5981 -2.0530 0.0000 C 0 0 0 0 0 0 0 0 0 0 0 0<br> 5.4641 -3.5530 0.0000 C 0 0 0 0 0 0 0 0 0 0 0 0<br> 7.8600 -2.5530 0.0000 C 0 0 0 0 0 0 0 0 0 0 0 0<br> 4.5981 -3.0530 0.0000 C 0 0 0 0 0 0 0 0 0 0 0 0<br> 4.5981 1.5670 0.0000 H 0 0 0 0 0 0 0 0 0 0 0 0<br> 3.1215 1.5547 0.0000 H 0 0 0 0 0 0 0 0 0 0 0 0<br> 3.5200 0.8644 0.0000 H 0 0 0 0 0 0 0 0 0 0 0 0<br> 2.8660 2.3270 0.0000 H 0 0 0 0 0 0 0 0 0 0 0 0<br> 4.2690 2.7570 0.0000 H 0 0 0 0 0 0 0 0 0 0 0 0<br> 3.9875 0.0547 0.0000 H 0 0 0 0 0 0 0 0 0 0 0 0<br> 4.3860 -0.6356 0.0000 H 0 0 0 0 0 0 0 0 0 0 0 0<br> 3.4860 3.9470 0.0000 H 0 0 0 0 0 0 0 0 0 0 0 0<br> 2.8660 4.5670 0.0000 H 0 0 0 0 0 0 0 0 0 0 0 0<br> 2.2460 3.9470 0.0000 H 0 0 0 0 0 0 0 0 0 0 0 0<br> 1.6900 2.9840 0.0000 H 0 0 0 0 0 0 0 0 0 0 0 0<br> 1.4631 2.1370 0.0000 H 0 0 0 0 0 0 0 0 0 0 0 0<br> 2.3100 1.9101 0.0000 H 0 0 0 0 0 0 0 0 0 0 0 0<br> 7.4690 -1.1589 0.0000 H 0 0 0 0 0 0 0 0 0 0 0 0<br> 4.0611 -1.7430 0.0000 H 0 0 0 0 0 0 0 0 0 0 0 0<br> 5.4641 2.0670 0.0000 H 0 0 0 0 0 0 0 0 0 0 0 0<br> 7.4690 -3.9470 0.0000 H 0 0 0 0 0 0 0 0 0 0 0 0<br> 5.4641 -4.1730 0.0000 H 0 0 0 0 0 0 0 0 0 0 0 0<br> 8.4800 -2.5530 0.0000 H 0 0 0 0 0 0 0 0 0 0 0 0<br> 4.0611 -3.3630 0.0000 H 0 0 0 0 0 0 0 0 0 0 0 0<br> 1 8 1 0 0 0 0<br> 1 10 1 0 0 0 0<br> 2 5 1 0 0 0 0<br> 2 34 1 0 0 0 0<br> 3 6 1 0 0 0 0<br> 3 7 1 0 0 0 0<br> 3 23 1 0 0 0 0<br> 4 13 1 0 0 0 0<br> 4 17 1 0 0 0 0<br> 4 35 1 0 0 0 0<br> 5 6 1 0 0 0 0<br> 5 8 1 0 0 0 0<br> 5 19 1 0 0 0 0<br> 6 20 1 0 0 0 0<br> 6 21 1 0 0 0 0<br> 7 11 1 0 0 0 0<br> 7 12 1 0 0 0 0<br> 7 22 1 0 0 0 0<br> 8 24 1 0 0 0 0<br> 8 25 1 0 0 0 0<br> 9 10 1 0 0 0 0<br> 9 13 2 0 0 0 0<br> 9 14 1 0 0 0 0<br> 10 15 2 0 0 0 0<br> 11 26 1 0 0 0 0<br> 11 27 1 0 0 0 0<br> 11 28 1 0 0 0 0<br> 12 29 1 0 0 0 0<br> 12 30 1 0 0 0 0<br> 12 31 1 0 0 0 0<br> 13 16 1 0 0 0 0<br> 14 17 2 0 0 0 0<br> 14 32 1 0 0 0 0<br> 15 18 1 0 0 0 0<br> 15 33 1 0 0 0 0<br> 16 18 2 0 0 0 0<br> 16 36 1 0 0 0 0<br> 17 37 1 0 0 0 0<br> 18 38 1 0 0 0 0<br>M END
Compound Name
Pindolol
CID
4828
Molecular Formula
C14H20N2O2
Molecular Weight
248
No. Hydrogen Bond Acceptors
3
No. Hydrogen Bond Donors
3
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