High throughput-solid phase extraction coupled with tandem mass spectrometry (HT-SPE-MS/MS, Agilent brand name RapidFire) is an automated system developed in last decade. The typical instrument time per sample on RapidFire is 10-30 seconds, which is several folds faster than chromatography based approaches. RapidFire system has been successfully used for in vitro assays, however, its utilization for the analysis of biological samples could be challenging. Currently industries and clinical organizations are very interested in using this innovative platform for clinical studies and therapeutic drugs monitoring, however a number of challenging issues (matrix effect, carryover and specificity) have yet to be rigorously evaluated.As a proof-of-concept study, we evaluated the feasibility of utilizing RapidFire for analysis of three metabolically related compounds, bupropion, hydroxyl-bupropion and thero-bupropion, in human plasma from a clinical trial on bioequivalence and food effect of bupropion. Critical bioanalytical validation parameters such as the matrix effect, specificity, accuracy, precision, analytical range and carry-over were systematically investigated. The analytical method (plasma sample preparation, RapidFire, MS/MS methods) was developed and validated according to the requirements of the FDA Bioanalytical Method Validation Guidance. In the second stage, we developed a RapidFire method for quantitating an anti-coagulant drug in human plasma and implemented the method to support review and surveillance activities at FDA.The analytical method for bupropion and its two major metabolites was validated according to the requirements of the Bioanalytical Method Validation Guidance and was determined to be accurate, precise, specific and linear over the corresponding analytical ranges. Comparable accuracy, precision, linearity, and sensitivity were achieved with analysis speeds 20 to 30-folds faster 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. Overall, the data from the proof-of-concept study indicate that the RapidFire system has significant potential for advancing high-throughput in vivo bioanalysis, e.g. bioavailability, bioequivalence and food effect studies. In summary, HT-SPE-MS/MS system is positioned to support review, surveillance, guidance and advance regulatory readiness.

Advanced robotics coupled with a mass spectrometry platform for clinical studies: Proof-of-concept to support review and surveillance

254th National Meeting (2017)

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