High throughput-solid phase extraction coupled with tandem mass spectrometry (HT-SPE-MS/MS, Agilent RapidFire) is an automated non-chromatographic system developed in last decade. The typical turn-a-round time per sample on RapidFire is 10-20 seconds, which is several times faster than chromatography based approaches. Due to lack of chromatographic separation, it is very crucial to optimize sample preparation methodology to manage the matrix effect when implementing bio-analytical analysis using the RapidFire system. In the current study, we systematically compare three sample preparation methods for analysis of three model drugs in human plasma.<br/>        The three different methods used to process human plasma samples were simplified liquid extraction (SLE), phospholipid deprivation filter (PLD), and enhanced matrix removal—lipid dispersive cleanup (EMR).  The analytical method (including plasma sample preparation, RapidFire and MS/MS methods) was developed and validated according to the requirements of the FDA Bioanalytical Method Validation Guidance.<br/>        We have successfully implemented PLD to process plasma samples for the RapidFire-MS/MS analysis of bupropion and its three metabolites, as well as a second model drug.  Comparable accuracy, precision, linearity, and sensitivity were achieved with analysis speeds 20 to 30-folds higher than conventional UPLC-MS method. Quantitation data for test compounds obtained from RapidFire were found to correlate well with the BE patient data from a validated UPLC-MS bupropion method. However, plasma processed using PLD and EMR still exhibited a strong matrix effect on the third model drug. A possible reason is that these methods might not be able to efficiently remove the other endogeneous compounds causing matrix effect. SLE, a plasma sample processing approach using a different extracting mechanism from PLD, was shown to be the best method to manage the matrix effect for analysis of the third model drug on RapidFire. In addition to optimizing sample processing methods for RapidFire-MS/MS, and the analysis of samples from three clinical studies, our work has allowed us to develop a novel decision tree for selection of plasma sample processing method to rapidly respond to clinical biopharmaceutics issues.<br/>

Determination of the best approach for processing human plasma samples to manage the matrix effect for analysis of three model drugs using RapidFire-MS/MS system

254th National Meeting (2017)

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