High Performance Liquid Chromatography and Photodiode Array Detection of Phenylpropanoids and Benzoates inRubus Protoplasts Elicited by Kinetin

Abstract

Reverse‐phase high performance liquid chromatography with photodiode array detection (PAD) has been developed for simultaneous separation, identification and quantification of picomoles of phenylalanine ammonia‐lyase (PAL) reaction products. Separation has been achieved in 30 min on a reverse‐phase silica column using methanol:water gradient elution, and PAD provided sensitive and specific determination of phenolics. The method has been applied to biological samples to study kinetin‐induced changes in phenol pathways within protoplasts of Rubus fruticosus (raspberry). © 1997 by John Wiley & Sons, Ltd.

High performance liquid chromatography and photodiode array detection of ferulic acid in Rubus protoplasts elicited by O-glycans from Fusarium sp. M7-1.

“…The presence of a PAL complex directing the flow of secondary compounds after hormone or elicitor treatment has been reported previously. The enzymes of branch pathways for phenylpropanoids, benzoates, or for flavonoids are known to be differentially expressed in response to environmental factors (Habereder et al, 1989;Funk & Brodelius, 1992;Chevolot et al, 1997). Likewise, specific enzymes, such as 3-O-methyltransferases acting at the terminal step in the biosynthesis of phenylpropanoids or of isoflavonoids, increased in response to elicitor or to pathogen (Busam et al, 1997;Grimmig & Matern, 1997;Preisig et al, 1991).…”

“…HPLC-PAD analysis. HPLC analyses were carried out as described in Chevolot et al (1997) but using a Kontron Liquid Chromatograph (Kontron Instruments, Milano, Italy) instead of a Waters one. The system encompassed a manual Rheodyne model 9125 injector, an HPLC 325 pump system, and a 440 photodiode array detector (PAD).…”

“…HPLC-PAD investigations of PAL extracts prepared from elicited protoplasts or from controls were run. Typical HPLC profiles of standards with detection at 256 and 310 nm were firstly achieved as in Chevolot et al (1997).…”