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The minor phenolic constituents of Cyclopia pubescens Eckl. Zeyh. are unknown and one dimensional (1D) liquid chromatography (LC) is unable to provide sufficient separation. A two-dimensional (2D) LC method incorporating normal-phasehigh performance countercurrent chromatography (NP-HPCCC) in the first dimension ( D) and reversed-phase ultra-high-performance liquid chromatography (RP-UHPLC) as the second dimension ( D) was developed. The analytical HPCCC method was subsequently scaled up to semi-preparative mode and fractions poo