Separation and quantification of basic hydroxy cinnamic amides and hydroxy cinnamic acids by reversed phase high performance liquid chromatography
Ponchet, M.; Martin Tanguy, J.; Poupet, A.; Marais, A.; Beck, D.
Journal of Chromatography 240(2): 397-404
A method for the separation of basic hydroxycinnamic amides and hydroxycinnamic acids by high-performance liquid chromatography is described. N-p-coumaryl-, N-caffeyl- and N-ferulylputrescine, N-p-coumaryl-, N-caffeyl- and N-ferulylspermidine and p-coumaric, caffeic, ferulic and sinapic acids were chromatographed on a .mu.Bondapak C18 reversed-phase column (particle size 9 .mu.m) with different methanol-water gradients as the mobile phase. It is possible with this high-resolution and reproducible method to assay biological samples containing > 10-5 M of hydroxycinnamic amides, using either p-coumaric or ferulic acid as the internal standard; this was demonstrated for tobacco [Nicotiana tabacum] extracts.