In this study, a water pre-treatment method for the separation and purification of an anticancer agent paclitaxel from plant cell cultures was developed. When the methanol extract obtained by biomass extraction was pre-treated with water, a high yield (>99%) of high-purity (similar to 12.1%) paclitaxel was observed within a short period of time (similar to 10 min). In addition, in the follow-up pre-purification process, using a crude extract obtained by water pre-treatment, high-yield (similar to 87.9%), high-purity (50% or higher) paclitaxel was obtained in a short operating time (similar to 2.9 h). Thus, the efficiency of the separation and purification process for paclitaxel was improved dramatically by water pre-treatment, particularly in terms of the reduction of operating times and the simplification of processes. (C) 2016 Elsevier Ltd. All rights reserved.