Phree Phospholipid Removal Solutions - Phenomenex
Phree Phospholipid Removal Solutions - Phenomenex
Phree Phospholipid Removal Solutions - Phenomenex
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One Quick Method, Three Big Advantages12Remove Proteins that can clog HPLC columnsEliminate <strong>Phospholipid</strong>s that cause ion suppressionand increase your column lifetime!3No Method Development; one method for acids,bases, and neutralsRemove ProteinsSolvent Shielding Technology preventsdripping of organic solvent, allowing forprotein precipitation within the <strong>Phree</strong><strong>Phospholipid</strong> <strong>Removal</strong> Product.Eliminate <strong>Phospholipid</strong>sThe <strong>Phree</strong> sorbent selectively removesphospholipids from precipitated plasmasamples.Proteins<strong>Phospholipid</strong>sTarget Analyte© 2013 <strong>Phenomenex</strong>, Inc. All rights reserved.2USA Tel: 310-212-0555 | Canada Tel: 800-543-3681 | Puerto Rico Tel: 800-541-4752<strong>Phenomenex</strong>| Fax: 310-328-7768 Email: info@phenomenex.com Web: www.phenomenex.com
<strong>Phospholipid</strong> <strong>Removal</strong> <strong>Solutions</strong>Be <strong>Phree</strong> of <strong>Phospholipid</strong>s and Proteins“We have demonstrated that <strong>Phree</strong> <strong>Phospholipid</strong><strong>Removal</strong> Plates resulted in >98 % decrease in thepeak area of the six phospholipids we monitoredin rat plasma compared to traditional protein crash.We highly recommend the use of these plates forhigh throughput LC-MS/MS bioanalytical samplepreparation.”Nina KhoshabaWIL Research3 Steps Above Traditional Protein Precipitation.............................pp. 4-5An Improved <strong>Phospholipid</strong> <strong>Removal</strong> Solution.................................pp. 6-7<strong>Phree</strong> Yourself of Method Development.........................................pp. 8-9Technical Support...............................................................................p. 10Ordering Information..........................................................................p. 11USA Tel: 310-212-0555 | Canada Tel: 800-543-3681 | Puerto Rico Tel: 800-541-4752<strong>Phenomenex</strong>| Fax: 310-328-7768 Email: info@phenomenex.com Web: www.phenomenex.com3
3 Steps Above Traditional Protein Precipitation1. Protein Precipitation Does Not Remove <strong>Phospholipid</strong>sEndogenous 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 <strong>Phospholipid</strong> ProfileProtein Precipitation vs. <strong>Phree</strong> <strong>Phospholipid</strong> <strong>Removal</strong> 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 <strong>Phree</strong><strong>Phree</strong> extracted plasmaProtein precipitated plasma0 1 23 4 5 6 7 8 9 1011 minApp ID 20012<strong>Phospholipid</strong> profile monitored using m/z 184-1844USA Tel: 310-212-0555 | Canada Tel: 800-543-3681 | Puerto Rico Tel: 800-541-4752<strong>Phenomenex</strong>| Fax: 310-328-7768 Email: info@phenomenex.com Web: www.phenomenex.com
<strong>Phospholipid</strong> <strong>Removal</strong> <strong>Solutions</strong>2. 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 observed<strong>Phree</strong> 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 Zone<strong>Phospholipid</strong>s m/z 184-184Amoxapine was infused post-column to establish an ion suppression/enhancement profile with both protein precipitatedplasma (left) and <strong>Phree</strong> extracted plasma (right), showing that <strong>Phree</strong> can successfully reduce ion suppression.Amoxapine m/z 314-2713. Maximize Sensitivity and Column Lifetime<strong>Phospholipid</strong>s 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 <strong>Phree</strong>Extracted PlasmaDiclofenac in ProteinPrecipitated Plasma100000<strong>Phospholipid</strong> 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 <strong>Phree</strong> extracted plasma were made.USA Tel: 310-212-0555 | Canada Tel: 800-543-3681 | Puerto Rico Tel: 800-541-4752<strong>Phenomenex</strong>| Fax: 310-328-7768 Email: info@phenomenex.com Web: www.phenomenex.com5
An Improved <strong>Phospholipid</strong> <strong>Removal</strong> SolutionRemove All Classes of <strong>Phospholipid</strong>sLysophosphatidylcholines and phosphatidylcholines both contribute to matrix effects. Remove allclasses of phospholipids using <strong>Phree</strong> <strong>Phospholipid</strong> <strong>Removal</strong> Products.1.5e6Intensity, cpsIntensity, cps1.4e61.5e61.3e61.4e61.2e61.3e61.1e61.2e61.0e61.1e69.0e51.0e68.0e59.0e57.0e58.0e56.0e57.0e55.0e56.0e54.0e55.0e53.0e54.0e52.0e53.0e51.0e52.0e501.0e5acetonitrile with 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 MethanolSupelco ® HybridSPE ® 60Plasma Cleanup: 100 µL plasma plus 300 µL<strong>Phenomenex</strong> ® Gradient: Time (min) % B<strong>Phree</strong>0 600.5 95Agilent ® Captiva NDLipids15.5 9515.51 60Flow Rate: 400 µL/minDetection: Mass Spectrometer (MS) @ 425 °C;184 amuTemperature: 22 °C0 12 3 4 5 6 7 8 9 10 min00 12 3 4 5 6 7 8 9 10 min~ 25x Zoom4.8e4Intensity, cpsIntensity, cps4.4e44.8e44.0e44.4e43.6e44.0e43.2e43.6e42.8e43.2e42.4e42.8e42.0e42.4e41.6e42.0e41.2e41.6e480001.2e44000800004000<strong>Phree</strong> is most effective atremoving all classes of phospholipids1 2 3 4 5 6 7 8 9 10 minApp ID 200150<strong>Phospholipid</strong> profile monitored1using m/z2184-1843 4 5 6 7 8 9 10 minAgilent is a registered trademark of Agilent Technologies Inc. Captiva is a trademark of Agilent Technologies, Inc.Supelco and HybridSPE are registered trademarks of Sigma-Aldrich Co.Comparative separations may not be representative of all applications.6USA Tel: 310-212-0555 | Canada Tel: 800-543-3681 | Puerto Rico Tel: 800-541-4752<strong>Phenomenex</strong>| Fax: 310-328-7768 Email: info@phenomenex.com Web: www.phenomenex.com
<strong>Phospholipid</strong> <strong>Removal</strong> <strong>Solutions</strong>Extended <strong>Phospholipid</strong> CapacityThe <strong>Phree</strong> sorbent has an extended capacity for phospholipids, allowing you to load up to 400 µL ofplasma without significant breakthrough of phospholipids.Intensity, cps200464.6e54.4e54.2e54.0e53.8e53.6e53.4e53.2e53.0e52.8e52.6e52.4e52.2e52.0e51.8e51.6e51.4e51.2e51.0e58.0e46.0e44.0e42.0e41200 1 2 3 4 5 6 7 8 9 10 11 min<strong>Phospholipid</strong> profile monitored using m/z 184-184~ 40x ZoomPlasma Cleanup: 400 µL plasma plus 1.2 mLacetonitrile with 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<strong>Phenomenex</strong> ® <strong>Phree</strong> Waters ® Ostro % B6095956060Flow Rate: 400 µL/minDetection: Mass Spectrometer (MS) @ 425 °C;184 amuTemperature: 22 °CApp ID 20046Waters Corporation states that the true maximum recommended plasma volume that can be processed using the Ostro plate is 350 µL due to well volume limitations.Ostro is a trademark of Waters Corporation. Comparative separations may not be representative of all applications.USA Tel: 310-212-0555 | Canada Tel: 800-543-3681 | Puerto Rico Tel: 800-541-4752<strong>Phenomenex</strong>| Fax: 310-328-7768 Email: info@phenomenex.com Web: www.phenomenex.com7
<strong>Phree</strong> Yourself of Method DevelopmentOne Method for Acids, Bases, and Neutrals1. DispensePlasma into the <strong>Phree</strong> tube or 96-well plate.2. AddOrganic solvent directly into the plasmasample.3. Mix4. FilterUsing a centrifuge, vacuum manifold,or positive pressure system.For complete method details and optimization tips,visit www.phenomenex.com/<strong>Phree</strong>One easy method for tubesand 96-well plates! Easilytransfer methods betweenthe two formats.Recommended Organic SolventsSolvent Solvent Volume Plasma VolumeAcetonitrile with 1 % Formic acid 300 µL 100 µLMethanol with 1 % Formic acid 400 µL 100 µLRecommended Plasma Loading CapacitiesFormatVolume96-well plates 25 - 400 µLTubes 25 - 200 µLTip<strong>Phenomenex</strong> accessories pair perfectly with<strong>Phree</strong> <strong>Phospholipid</strong> <strong>Removal</strong> Products. Go topage 11 for a complete list of accessories.8USA Tel: 310-212-0555 | Canada Tel: 800-543-3681 | Puerto Rico Tel: 800-541-4752<strong>Phenomenex</strong>| Fax: 310-328-7768 Email: info@phenomenex.com Web: www.phenomenex.com
<strong>Phospholipid</strong> <strong>Removal</strong> <strong>Solutions</strong>High Recoveries of Target AnalytesThe <strong>Phree</strong> general protocol has been developed to produce high recoveries of acids, bases,and neutrals while simultaneously removing all classes of phospholipids.Absolute Recoveries of Acids, Bases, and Neutrals100%90%80%70%Absolute Recovery (%)60%50%40%30%20%10%DiclofenacNaproxenPrednisoneAcetaminophenMetoprololAmitriptylineAcidsNeutralsBases0%AnalyteRecovery data was obtained by calculating the average absolute recoveries of analytes extracted from 3 <strong>Phree</strong> plates.USA Tel: 310-212-0555 | Canada Tel: 800-543-3681 | Puerto Rico Tel: 800-541-4752<strong>Phenomenex</strong>| Fax: 310-328-7768 Email: info@phenomenex.com Web: www.phenomenex.com9
Sample Preparation Specialists areReady to Assist YouTwo Levels of Method Development SupportLevel 1Contact one of our dedicated sample preparationspecialists for immediate method developmentassistance.Level 2Send your sample to our analytical servicesgroup for custom method development. Visitwww.phenomenex.com/PhenoLogix formore information.Get StartedContact your Sample Preparation Specialist:By phone: 310-212-0555By email: Support@phenomenex.com“<strong>Phenomenex</strong>’s prompt support isvery important for us to achieveour work. I thank them for theirpersistent support and innovativeproducts.”Allena JiWyeth10USA Tel: 310-212-0555 | Canada Tel: 800-543-3681 | Puerto Rico Tel: 800-541-4752<strong>Phenomenex</strong>| Fax: 310-328-7768 Email: info@phenomenex.com Web: www.phenomenex.com
<strong>Phospholipid</strong> <strong>Removal</strong> <strong>Solutions</strong>Order <strong>Phree</strong> Now!Ordering Information<strong>Phree</strong> <strong>Phospholipid</strong> <strong>Removal</strong> ProductsPart No. Description Unit Price8B-S133-TAK <strong>Phree</strong> <strong>Phospholipid</strong> <strong>Removal</strong> 1 mL Tube 100/box $1848E-S133-TGB <strong>Phree</strong> <strong>Phospholipid</strong> <strong>Removal</strong> 96-Well Plates 2/box 366AccessoriesCollection Plates (deep well, polypropylene)AH0-7192 Strata ® 96-Well Collection Plate 350 µL/well 50/pk $ 250AH0-7193 Strata 96-Well Collection Plate 1 mL/well 50/pk 250AH0-7194 Strata 96-Well Collection Plate 2 mL/well 50/pk 290AH0-8635 Strata 96-Well Collection Plate, 2 mL Square/Round-Conical 50/pk 290AH0-8636 Strata 96-Well Collection Plate, 2 mL Round/Round, 8 mm 50/pk 321AH0-7279 Strata 96-Well Collection Plate, 1 mL/well Round, 7 mm 50/pk 210Sealing MatsAH0-8597 Sealing Mats, Pierceable, 96-Square Well, Silicone 50/pk $ 255AH0-8598 Sealing Mats, Pre-Slit, 96-Square Well, Silicone 50/pk 280AH0-8631 Sealing Mats, Pierceable, 96-Round Well 7 mm, Silicone 50/pk 255AH0-8632 Sealing Mats, Pre-Slit, 96-Round Well 7 mm, Silicone 50/pk 280AH0-8633 Sealing Mats, Pierceable, 96-Round Well 8 mm, Silicone 50/pk 255AH0-8634 Sealing Mats, Pre-Slit, 96-Round Well 8 mm, Silicone 50/pk 280AH0-7362 Sealing Tape Pad 10/pk 109Vacuum ManifoldsAH0-6023* SPE 12-Position Vacuum Manifold Set, for tubes ea $ 634AH0-6024* SPE 24-Position Vacuum Manifold Set, for tubes ea 874AH0-8950 Strata 96-Well Plate Manifold, Universal with Vacuum Gauge ea 1,109*Manifolds include: Vacuum-tight glass chamber, vacuum gauge assembly, polypropylene lid with gasket, male andfemale luers and yellow end plugs, stopcock valves, collection rack assemblies, polypropylene needles, lid supportlegs. Waste container included with 12-positive manifold.If <strong>Phree</strong> <strong>Phospholipid</strong> <strong>Removal</strong> products do not perform as well or better thanyour current phospholipid removal product, send in your comparative datawithin 45 days and keep the <strong>Phree</strong> <strong>Phospholipid</strong> <strong>Removal</strong> product for FREE.TipFor more information about <strong>Phenomenex</strong>sample preparation products, visitwww.phenomenex.com/sampleprepinfoTerms and ConditionsSubject to <strong>Phenomenex</strong> Standard Terms and Conditions, which may be viewed at www.phenomenex.com/TermsAndConditions.TrademarksStrata and Kinetex are registered trademarks of <strong>Phenomenex</strong>. <strong>Phree</strong> and Solvent Shielding Technology are trademarks of <strong>Phenomenex</strong>.Agilent is a registered trademark, and Captiva is a trademark of Agilent Technologies, Inc. Supelco and HybridSPE are registered trademarks of Sigma-Aldrich Co.Waters is a registered trademark, and Ostro is a trademark of Waters Corporation.Disclaimer<strong>Phenomenex</strong> is in no way affiliated with Agilent Technologies, Sigma Aldrich, or Waters Corporation. Comparative separations may not be representative of all applications.The opinions stated herein are solely those of the speaker and not necessarily those of any company or organization.© 2013 <strong>Phenomenex</strong>, Inc. All rights reserved.USA Tel: 310-212-0555 | Canada Tel: 800-543-3681 | Puerto Rico Tel: 800-541-4752<strong>Phenomenex</strong>| Fax: 310-328-7768 Email: info@phenomenex.com Web: www.phenomenex.com11
PRSRT STDU.S. POSTAGEPAIDTORRANCE, CAPERMIT NO. 185<strong>Phospholipid</strong> <strong>Removal</strong> <strong>Solutions</strong>BR13070413_usAustraliat: 02-9428-6444f: 02-9428-6445auinfo@phenomenex.comAustriat: 01-319-1301f: 01-319-1300anfrage@phenomenex.comBelgiumt: 02 503 4015 (French)t: 02 511 8666 (Dutch)f: +31 (0)30-2383749beinfo@phenomenex.comCanadat: (800) 543-3681f: (310) 328-7768info@phenomenex.comDenmarkt: 4824 8048f: +45 4810 6265nordicinfo@phenomenex.comFinlandt: 09 4789 0063f: +45 4810 6265nordicinfo@phenomenex.comFrancet: 01 30 09 21 10f: 01 30 09 21 11franceinfo@phenomenex.comGermanyt: 06021-58830-0f: 06021-58830-11anfrage@phenomenex.comIndiat: 040-3012 2400f: 040-3012 2411indiainfo@phenomenex.comIrelandt: 01 247 5405f: +44 1625-501796eireinfo@phenomenex.comItalyt: 051 6327511f: 051 6327555italiainfo@phenomenex.comLuxembourgt: +31 (0)30-2418700f: +31 (0)30-2383749nlinfo@phenomenex.comMexicot: 001-800-844-5226f: 001-310-328-7768tecnicomx@phenomenex.comThe Netherlandst: 030-2418700f: 030-2383749nlinfo@phenomenex.comNew Zealandt: 09-4780951f: 09-4780952nzinfo@phenomenex.comNorwayt: 810 02 005f: +45 4810 6265nordicinfo@phenomenex.comPuerto Ricot: (800) 541-HPLCf: (310) 328-7768info@phenomenex.comSwedent: 08 611 6950f: +45 4810 6265nordicinfo@phenomenex.comUnited Kingdomt: 01625-501367f: 01625-501796ukinfo@phenomenex.comUnited Statest: (310) 212-0555f: (310) 328-7768info@phenomenex.comAll other countries:Corporate Office USAt: (310) 212-0555f: (310) 328-7768info@phenomenex.comwww.phenomenex.com<strong>Phenomenex</strong> products are available worldwide. For the distributor in your country,contact <strong>Phenomenex</strong> USA, International Department at international@phenomenex.com411 Madrid AvenueTorrance, CA 90501-1430USA
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