W529posterSBS2011 Shieh MassSpecAssays
Ultrasonic Fluid Processing Improves Triolein Measurement Accuracy in Mass Spectrometry Assays
Lauren Frick1, Jim O’Keefe2, Jennifer Rossi1 and Jean Shieh2. 1BIOCIUS Life Sciences (now Agilent Technologies); 2Microsonic Systems.
Development of therapeutics for obesity is the focus of many pharmaceutical research and development programs, particularly the identification of potential inhibitors of the triacylglycerol (TAG) synthesis pathway. The reaction catalyzed by the enzyme diaceylglycerol aceyl transferase (DGAT) in the TAG pathway produces triolein, and because triolein remains in the lipid bilayer of the cell membrane rather than in the assay buffer making it difficult to accurately measure the compound activity and to reliably detect triolein in the mass spectrometer. In the protocol described in this poster, we used the Microsonic Systems HENDRIX SM100 ultrasonic fluid processor to release triolein from the membrane bilayer, and then used the BIOCIUS RapidFire® Mass Spectrometry (RF-MS) platform to accurately detect and quantify native compounds. We compared the results of various HENDRIX SM100 settings to determine the optimal operating conditions and achieved a 50% improvement in the triolein signal-to-noise ratio in the RF-MS assay.
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