A Parallel Micro Turbulent Flow Chromatography-Tandem Mass Spectrometry Method for the Analysis of a Pharmaceutical Compound in Plasma

Turbulent flow chromatography coupled to tandem mass spectrometry (TFC-MS-MS) has been widely used within bioanalysis, because of the ability to inject directly neat biological samples with no prior pretreatment. TFC-MS-MS removes the need for time consuming sample preparation procedures such as protein precipitation, liquid-liquid extraction or solid-phase extraction. There are two standard configurations that are commonly adopted for the use of TFC, namely fast elute and focus mode. In this paper, a new micro TFC column which has the advantages of reducing solvent consumption and improving mass sensitivity, was used to develop a fast elute method. A wide range of pharmaceutical compounds with various physicochemical properties was used to assess the system performance. Carry-over has often been identified as a potential source of inaccuracy in a fast elute mode. This paper shows how by using a systematic approach, carryover was eliminated. The parameters influencing the robustness of the micro TFC column are also discussed. This method was fully validated for a Pfizer development compound in human plasma using a new TFC parallel platform allowing two systems to be operated in parallel. Multiplexing the sample analysis allowed a 2-fold increase in throughput and provided an efficient use of the mass spectrometer.