Introduction.

Liquid chromatography (LC) is a widely used technique for the separation, isolation, and purification of chemical compounds present in mixtures. The direct coupling of biological detection to liquid chromatography may facilitate the chemical and biological characterization of active compounds.

Methods.

In previous years, a system was optimized that combined the advantages of LC separation with classical systems for analyzing pharmacological activity in isolated perifused or perfused organs.¹ Thus, a hydro-ethanolic extract of Stevia rebaudiana Bertoni (Asteraceae) was studied through a method based on medium-pressure liquid chromatography separation (MPLC) coupled directly to a living superfused organ cascade as a quadruple biosensor.¹

In this poster communication, the potential uses of and perspectives in organic chemistry and pharmacology on this direct coupling of biological detection to an MPLC system are presented.

Results.

Rebaudioside N, a minor natural product produced from Stevia rebaudiana Bertoni, was identified as the compound responsible for the contractile activity detected in the active fraction of the initial hydro-ethanolic extract.¹ The isolated rebaudioside N contracted the smooth muscle present in portions of rat ilea. This stevioside was structurally identified using mass spectrometry.¹

In the process of pharmacological characterization of new active compounds, this type of methodology reduced the investigation time, number of animals slaughtered, use of organic solvents, and associated expenses.¹

Conclusions.

This direct combination of liquid chromatography with a perfusion system will allow bioactive compounds present in mixtures derived from extracts of natural origin or chemical synthesis (for example, combinatorial chemistry) to be isolated and pre-characterized.

References.

¹ Campuzano-Bublitz et al., Naunyn-Schmiedeberg's Arch Pharmacol2018, 391, 9–16.