Phree Phospholipid Removal Solutions - Phenomenex

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3 Steps Above Traditional Protein Precipitation1. Protein Precipitation Does Not Remove PhospholipidsEndogenous phospholipids are a primary source of ion suppression and resulting matrix effects inbioanalytical LC/MS work. Ion suppression caused by the presence of phospholipids can result in:• irreproducible results• quantitation issues• loss in method sensitivity• matrix to matrix biasTotal Phospholipid ProfileProtein Precipitation vs. Phree Phospholipid Removal ProductsIntensity, cps200114.5e64.0e63.5e63.0e62.5e62.0e6Plasma Cleanup: 100 µL plasma plus 300 µL acetonitrilewith 1 % formic acidColumn: Kinetex ® 2.6 µm C18 100 ÅDimensions: 50 x 2.1 mmPart No.: 00B-4462-ANMobile Phase: A: 0.1% Formic acid in WaterB: 0.1% Formic acid in MethanolGradient: Time (min)00.515.515.5119.5% B6095956060Flow Rate: 400 µL/minDetection: Mass Spectrometer (MS) @ 425 °C;184 amuTemperature: 22 °C1.5e61.0e65.0e5App ID 2001100 1 23 4 5 6 7 8 9 1011 min~ 50x Zoom20012Intensity, cps9.0e48.5e48.0e47.5e47.0e46.5e46.0e45.5e45.0e44.5e44.0e43.5e43.0e42.5e42.0e41.5e41.0e45000.00.0Selectively removephospholipids with PhreePhree extracted plasmaProtein precipitated plasma0 1 23 4 5 6 7 8 9 1011 minApp ID 20012Phospholipid profile monitored using m/z 184-1844USA Tel: 310-212-0555 | Canada Tel: 800-543-3681 | Puerto Rico Tel: 800-541-4752Phenomenex| Fax: 310-328-7768 Email: info@phenomenex.com Web: www.phenomenex.com
Phospholipid Removal Solutions2. Reduce Ion SuppressionThe presence of phospholipids in plasma samples produces zones of ion suppression that correlateexactly with the phospholipid elution profile when analyzed via mass spectrometer (MS).1.2e61.0e6Protein Precipitated Plasma20013Ion suppressionis observedPhree Extracted Plasma200141.2e61.0e6Intensity, cps8.0e56.0e5Intensity, cps8.0e56.0e5Suppression zoneis eliminated4.0e52.0e5App ID 200134.0e52.0e5App ID 200140.00.2 0.6 1.0 1.4 1.8 2.2 2.6 min00 0.2 0.6 1.0 1.4 1.8 2.2 2.6 minSuppression ZonePhospholipids m/z 184-184Amoxapine was infused post-column to establish an ion suppression/enhancement profile with both protein precipitatedplasma (left) and Phree extracted plasma (right), showing that Phree can successfully reduce ion suppression.Amoxapine m/z 314-2713. Maximize Sensitivity and Column LifetimePhospholipids reduce the sensitivity of the MS signal and shorten column lifetime whenthey build up over time.Column Sensitivity after 250 Injections300000250000~5,000 µL plasma injected withoutsignificant loss of sensitivity200000Peak Area Counts150000Reduced sensitivity isimmediately observedDiclofenac in PhreeExtracted PlasmaDiclofenac in ProteinPrecipitated Plasma100000Phospholipid build up results insignificant loss of sensitivity5000000 50 100 150 200 250Injection Number (20 µL of Plasma)To assess the effect of phospholipid build up, repetitive 20 µL injections of diclofenac in protein precipitated plasma versus diclofenac in Phree extracted plasma were made.USA Tel: 310-212-0555 | Canada Tel: 800-543-3681 | Puerto Rico Tel: 800-541-4752Phenomenex| Fax: 310-328-7768 Email: info@phenomenex.com Web: www.phenomenex.com5
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Phree Phospholipid Removal Solutions - Phenomenex

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