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Phospholipid Removal Phree vs. Traditional PPT(Zoomed Scale)

Sample Preparation Details

Part No:8E-S133-TGB
Solid Phase Extraction (SPE) procedure:Note: The solvent volumes shown below are for a 30 mg bed mass. The solvent volumes will need to be adjusted for a smaller or larger bed mass.
Load:Dispense 300 uL ACN with 1% Formic acid into the wells of the plate.
Add 100 uL of plasma.
Mix by vortexing or aspiration.
Filter via centrifugation, vacuum, or positive pressure.
Final Prep and Analysis: Inject: 2 µL on HPLC / Mass Spectrometer (MS) @ 184 amu (425 °C)

Analytes:

Name Spiked
Conc.
Log P pKa % Rec %RSC (n=0)
1 Phospholipid Profile

Application Detail (App ID: 20012)

Part No:00B-4462-AN
Dimensions:50 x 2.1 mm ID
Elution Type:Gradient
Elution A:0.1% Formic Acid in Water
Elution B:0.1% Formic Acid in Methanol
Gradient Profile:
Step No. Time (min) Pct A Pct B
1 0 40 60
2 0.5 5 95
3 15.5 5 95
4 15.51 40 60
5 19.5 40 60

Flow Rate:400 µL/min
Col. Temp: ambient
Detection: Mass Spectrometer (MS) @ 184 amu (425°C)
Note:Extracted sample was prepped with Phenomenex Phree Phospholipid Removal Plate. Step 1: Add 300uL of 1% formic acid in acetonitrile directly to Phree plate. Step 2: Add 100uL of plasma directly to Phree plate. Vortex and extract. This was compared to standard protein precipitated plasma using the same volume of plasma and crash solvent

Analyte Details

Analyte Detail not available

1
Phospholipid Profile