Turbulent flow bioanalysis in drug metabolism and pharmacokinetics

Abstract

Turbulent flow chromatography (TFC) as an analytical technique was introduced in the mid-1990s for online sample processing in bioanalysis. Turbulent flow columns are packed with large particles permitting the use of high mobile phase linear velocities. Solute molecules travel in a uniform concentrated band in which large matrix biomolecules are effectively separated from lower molecular weight analytes by differential mass transfer effects. In addition, the use of various bonded stationary phases enhances selectivity in TFC separations. Originally, the turbulent flow column was used for both online sample processing and the analytical separation. This mode of operation has largely been replaced with a dual column arrangement, in which sample processing is done online with the turbulent flow column and a second analytical column is employed for separation. Within the pharmaceutical industry, certain laboratories now use TFC routinely for bioanalytical support of in vivo drug discovery pharmacokinetic studies. Validated TFC methods are also used to support GLP-compliant preclinical toxicokinetic studies and clinical trials. TFC has been shown to simplify bioanalytical sample preparation and reduce sample processing costs when compared with offline techniques such as SPE. TFC may be combined with multiplexing up to four HPLC systems to a single mass spectrometer to increase analytical throughput.