Abstract

Two dimensional thin layer chromatography on DIOL polar bonded stationary phase was performed to optimize the separation of some anioxidant phenolic compounds from Eupatorium cannabinum extracts. Ethyl-methyl ketone mixed with n-heptane and ethyl acetate mixed with n-heptane were used as non-aqueous mobile phases in normal phase separations (1st direction of development in 2D-HPTLC mode) and methanol mixed with water was used as a mobile phase in reversed phase (2nd direction of development in 2D-HPTLC mode). The plates were sprayed by use of Merck TLC sprayer using 2-(diphenylboryoxy)-ethylamine and PEG4000 (Merck, Darmstadt, Germany) or DPPH and photographed in Camag Cabinet UV lamp at 254 nm and 365 nm by use of Fuji 8 mpx camera. Satisfactory separations of antioxidant phenolic compounds in Eupatorium cannabinum extracts were obtained by use of optimized 2D-HPTLC systems.

Keywords: 2D-HPTLC, DIOL stationary phase, Eupatorium cannabinum, plant extracts, antioxidant activity, phenolics, DPPH reagent, camera, thin layer chromatography, flavonoid aglycones