In this study, two new polyacetylenes were isolated from the extract portion with ethyl acetate from Dolomiaea souliei roots by implementing various chromatography techniques, such as the use of silica gel under normal-phase conditions, MPLC, molecular sieving utilising Sephadex LH-20, and purification through semi preparative HPLC. Their structures were elucidated as (E)-[(Z)-deca-8-en-4,6-diynyl]-3-(4-hydroxy-3-methoxyphenyl) acrylate (1) and 6 R-(4E, 12E)-tetradeca-4,12-dien-8,10-diyne-1,6-diyldiacetate (2) by multiple spectroscopic techniques such as 1D and 2D NMR, UV, IR, and HR ESI MS.