High‐performance liquid chromatographic method for identification and quantification of two isomeric coumarinolignoids—cleomiscosin A and cleomiscosin B—in extracts of Cleome viscosa

A simple, accurate and reproducible reverse‐phase HPLC method has been developed for identification and quantification of two isomeric coumarinolignoids, cleomiscosin A and B in different extracts of the seeds of Cleome viscosa using photodiode array detection at 326 nm. Cleomiscosin A and B were separated on a Waters symmetry C 18 column (250 × 4.6 mm with 5.0 µm particle size) with an isocratic elution system composed of acetonitrile–methanol (1:2, v/v) and acetic acid–water (0.5:99.5, v/v) in the ratio of 40:60 (v/v). The calibration curves were linear ( r 2 > 0.997) in the concentration ranges of 20–100 µg/mL for both compounds. The limits of detection and quantification were 15 and 20 µg/mL for both cleomiscosin A and B. The intra‐ and inter‐day precisions were 3.68 and 2.22% for cleomiscosin A and 4.22 and 5.06% for cleomiscosin B. The recoveries measured at two different concentration levels varied from 98.03 to 110.06%. The method was used to identify and quantify cleomiscosins A and B in different extracts of Cleome viscosa seeds. Copyright © 2007 John Wiley & Sons, Ltd.