The invention belongs to the technical field of plant extraction and analysis, and particularly relates to an actinidia arguta root extract as well as an extraction and separation method and application thereof. The extract comprises the following 21 chemical components: 21 chemical components; the invention discloses a preparation method of 7-O-[beta-D-xylose pyranosyl-(1-6)-beta-D-glucopyranoside]-1, 2, 4, 5, 6, 7-O-[beta-D-xylose pyranosyl-(1-6)-beta-D-glucopyranoside]-1. The preparation method comprises the following steps: preparing 2, 8-dihydroxy-3-methoxy-ketone; 3-O-[beta-D-rhamnose-(1-6)-beta-D-galactopyranoside]-5, 7, 3 ', 4'-tetrahydroxy flavone, catechin, beta-sterol, quercetin, isoquercitrin, oleanolic acid, ursolic acid, 2alpha, 3beta-dihydroxy oleanane-12 alkene-28 acid, 2alpha, 3alpha, 24-trihydroxy ursolic acid and the like. The extract provided by the invention has a good application prospect in preparation of anti-cancer drugs for improving the advanced living standard of cancer patients.