Solid Phase Extraction (SPE)

Selective Depletion of Phospholipids in Bioanalysis using HybridSPE-PPT Technology

Oct 15 2009

Author: Craig Aurand and An Trinh on behalf of Supelco

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The approach of this study was to evaluate the performance of recently developed sample prep platform HybridSPE(TM) Precipitation(HybridSPE-PPT)for the purpose of matrix removal and analyte recovery from spiked plasma samples. The goal of the study was to demonstrate how the HybridSPE-PPT platform can increase bioanalytical throughput by both decreasing sample preparation time and improving chromatographic analysis and detection relative to traditional protein precipitation methods.

Often the major concern in developing bioanalytical methods is addressing the effect of endogenous biological matrix components on the detection of desired analytes. The impact of matrix effects in bioanalysis has
been well documented [1,2]. In the majority of cases, co-extracted interferences directly affect the quantitation of analytes due to ionization effects induced by the extracted matrix. This extracted matrix can impact the
chromatographic analysis, but more often is a result of chromatographic build up that leads to irregularities. Chromatographic build up of phospholipids is often observed when performing standard protein precipitation
techniques [3,5]. Adequate removal of phospholipids from the analytical column often result in the need for gradient elution under reversed-phase separations resulting in prolonged analysis. Even ballistic conditions
often used with modern UHPLC instruments are inadequate to remove phospholipids that a have accumulated on the column. In most cases gradient elution is not required for resolution of desired analytes, but instead
required only to elute extracted matrix from the analytical column.

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