The 8th Mass Spectrometry CVG Annual Symposium - Full Day Event!!!

"Analytical, Bioanalytical, Environmental, Proteomics, Forensic and Instrumental Topics "

September 23, 2009, Montreal, Canada

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Presentation No. 12 - A simple way to remove phospholipids from bioanalytical samples
Ben Yong1; David Jones1; Ritu Arora1, Paul Boguszewski2
1Varian, Inc., Lake Forest , CA, 2Varian, Inc., Essex Road, Church Stretton, UK

Novel Aspect: Phospholipid removal from protein precipitation in a 96 well plate.

Introduction: Bioanalytical analysis requires removal of matrix which interferes with LC/MS analysis. Phospholipid removal has received much attention in literature recently. Phospholipids can cause ion suppression, and shorten column lifetime. Removal of Phospholipids is generally achieved by SPE or liquid liquid extraction. A new development of lipid depleted protein precipitation allows the user to remove lipids with a simple general methodology, saving both costs and method development time. This filtration based, depletion approach gives improved cleanliness over protein precipitation, yet avoids additional method development or sample processing time.

Methods: Captiva NDLipids 96 well plates are packed with a non-drip membrane, a protein filtration membrane and a proprietary sorbent in order remove proteins and lipids. Typical precipitation conditions are 3:1 acidified MeOH to plasma although alternate solvents and ratios are also discussed. Samples are filtered under vacuum and analyzed by LC-MS/MS. Lipid removal is monitored by measuring the 184-184 MS transition and sample cleanliness monitored using post-column infusion.

Results: Good recoveries were achieved for a range of pharmaceutical compounds with a range of logP values. The protein and lipid removal are dependant on the exact precipitation conditions used as well as several other key parameters. Using these results we are pleased to present a set of generic conditions for protein precipitation on the Captiva NDLipids 96 well plates under which the majority of proteins and phospholipids can be removed.