Quantitative Nuclear Magnetic Resonance and High-Performance Thin-Layer Chromatography of Cupressuflavone as a Marker for Cupressus Species

Cupressuflavone is a major biflavonoid in many member of the family Cupressaceae and considered the main chemotaxonomic marker for Cupressus species. Despite the abundance of cupressuflavone, it has not been intensively investigated as compared to other biflavonoids. The current study illustrates, for the first time, a comparative analysis on the cupressuflavone content in five Cupressus hydroalcoholic extracts using novel, direct, simple, and time-saving analytical techniques namely quantitative nuclear magnetic resonance and normal-phase high-performance thin-layer chromatography. The analyzed samples were leaves and cones of Cupressus sempervirens var. horizontalis (Mill.) Loudon and C. sempervirens var. pyramidalis (O.Targ.Tozz.) Nyman, as well as leaves of C. macrocarpa Hartw. ex Gordon. The highest cupressuflavone content was detected in the leaves of Cupressus sempervirens var. horizontalis (0.958 g % dried powdered plant). In general, higher contents of cupressuflavone, more than 100 folds, were determined in the leaf samples. The results for the quantification of this chemical marker were similar by the two applied analytical methods. In addition, in vitro free radical scavenging activities of the studied Cupressus samples were evaluated.