Assay of 2,3-dihydroxybenzoic acid and related compounds in plant materials by high-performance liquid chromatography.
Salicylic acid and its putative biosynthetic precursors were assayed isocratically by RP-HPLC with UV detection at 280 nm. Optimum resolution was provided by an HPLC mobile phase consisting of MeOH-1% aqueous HOAc (40:60, v/v), at pH 4. Furthermore, for the analysis of 2,3-dihydroxybenzoic acid (2,3-DHBA) in Catharanthus roseus cell cultures after elicitation, a mobile phase consisting of acetonitrile-1% aqueous HCOOH containing 0.25% trichloroacetic acid (1:5, v/v), at pH 2, was used. The recovery for the free form of 2,3-DHBA was about 80% after a one-step extraction of the cells. The detection limit of 2,3-DHBA was 3 microg by using saligenin as an internal standard.
Muljono
R A
RA
Division of Pharmacognosy, Leiden/Amsterdam Center for Drug Research, Leiden University, The Netherlands.
Darsono
F L
FL
Scheffer
J J
JJ
Verpoorte
R
R
D030008:Catharanthus / Q000737:chemistry* / Q000166:cytology; D002478:Cells, Cultured; D002851:Chromatography, High Pressure Liquid / Q000379:methods*; D006863:Hydrogen-Ion Concentration; D062385:Hydroxybenzoates / Q000032:analysis*; D012680:Sensitivity and Specificity
D062385:Hydroxybenzoates; C009135:2,3-dihydroxybenzoic acid