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Turbulent flow chromatography TFC-tandem mass spectrometry supporting in vitro/vivo studies of NCEs in high throughput fashion.

By Verdirame, Maria; Veneziano, Maria; Alfieri, Anna; Di Marco, Annalise; Monteagudo, Edith; Bonelli, Fabio
From Journal of Pharmaceutical and Biomedical Analysis (2010), 51(4), 834-841. Language: English, Database: CAPLUS, DOI:10.1016/j.jpba.2009.10.005

Turbulent Flow Chromatog. (TFC) is a powerful approach for online extn. in bioanal. studies. It improves sensitivity and reduces sample prepn. time, two factors that are of primary importance in drug discovery. In this paper the application of the ARIA system to the anal. support of in vivo pharmacokinetics (PK) and in vitro drug metab. studies is described, with an emphasis in high throughput optimization. For PK studies, a comparison between acetonitrile plasma protein pptn. (APPP) and TFC was carried out. The authors’ optimized TFC methodol. gave better S/N ratios and lower limit of quantification (LOQ) than conventional procedures. A robust and high throughput anal. method to support hepatocyte metabolic stability screening of new chem. entities was developed by hyphenation of TFC with mass spectrometry. An in-loop diln. injection procedure was implemented to overcome one of the main issues when using TFC, that is the early elution of hydrophilic compds. that renders low recoveries. A comparison between off-line solid phase extn. (SPE) and TFC was also carried out, and recovery, sensitivity (LOQ), matrix effect and robustness were evaluated. The use of two parallel columns in the configuration of the system provided a further increase of the throughput.

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