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.

Determination of arachidonic acid by on-line solid-phase extraction HPLC with UV detection for screening of cytosolic phospholipase A2α inhibitors

“…However, with this material peak broadening occurred after only a few injections due to saturation of the column by binding of components present in the enzyme preparation, most likely proteins. In literature it has been reported that the so called turbulent flow chromatography is suitable for separating large molecules from low mass analytes [32]. The columns used in this type of chromatography consist of porous particles with sizes that generally range from 30 to 50 m. Operating at high flow rates of the mobile phase of about 4 mL/min for 1 mm internal diameter columns leads to turbulent flows in the column preventing large biomolecules to diffuse into the pores of the stationary phase.…”

“…After 2 min, the valve was automatically switched to position 2 and the analyte was back-flushed off the trap column onto the analytical column with the mobile phase delivered by pump B (acetonitrile/water/phosphoric acid (85%), 77:23:0.1, v/v/v) at a flow rate of 0.4 mL/min. A Waters UV detector 486 was used for detection at 200 nm and system control was performed by Waters Millennium [32] software. The temperature of the autosampler was kept at 10 • C. The analytical column was temperature controlled at 20 • C by a jetstream plus column oven from Waters.…”

Turbulent flow chromatography in bioanalysis: a review

“…Quick Elute Mode (Fig. ) is designed for ultra rapid, targeted analysis of small molecules, and was more commonly chosen for early methods using TurboFlow (Rudewicz, ). In the first published method using TurboFlow, Ayrton et al .…”

“…Quick Elute Mode (Fig. 3) is designed for ultra rapid, targeted analysis of small molecules, and was more commonly chosen for early methods using TurboFlow (Rudewicz, 2011). In the first published method using TurboFlow, Ayrton et al wileyonlinelibrary.com/journal/bmc a quantitative, Quick Elute Mode method for a proprietary isoquinoline compound in plasma for toxicokinetic studies.…”

Current Trends in Sample Treatment Techniques for Environmental and Food Analysis

“…The recent implementation of automated on-line extraction procedures has allowed fast sample clean-up in bioanalytical applications, and turbulent-flow (or TurboFlow) chromatography (TFC) appears as one of the most interesting ones in this area [102][103][104][105].…”