Biotechnological research to achieve and then increase anthocyanin production in callus tissue of R. hirta L., involved testing a number of growth media, modified with growth regulators. The evaluation of the growth of propagated tissues, their water contents and in particular their ability to produce anthocyanins, led to the development of an original two-phase growth system. This new system has the advantages of both growth-stimulating media and pigment production media. As a result of the research, callus tissue was obtained on a two-phase growth medium, which was made of modified Schenk-Hildebrandt medium (growth phase) and Miller's medium (production phase). The callus synthesised a 12-component anthocyanin compound complex, which constituted 2,18% of dry mass. This is a considerable amount compared to 0,28% in the natural plant. Phytochemical analysis (TLC, HPLC, PC, UV, 1H-NMR) of the anthocyanin complex isolated from callus produced with the two-phase system proved that the dominating compounds in the pigment complex were: cyanidin-3-0-(6-0-malonyl-b -D-glucopyranoside) and cyanidin-3-0-(b-D-glucopyranoside).