Recently, there has been a special research focus on the bioconversion of glycerol to 1,3-propanediol (1,3-PD) due to its significance in the chemical industry. However, the treatment and separation of fermentation broths is a great challenge. Currently, the reverse osmosis (RO) process is a reliable state-of-the-art technique for separation of biological solutions. This study (as the first to do so) investigated the feasibility of separation of 1,3-PD broths with the use of cellulose acetate (CA) membrane by the RO process. The experiments were carried out using the installation equipped with the plate module, under the transmembrane pressure (TMP) and temperature of 1 MPa and 298 K, respectively. It was found that the used membrane was suitable for broth separation. Indeed, it was noted that 1,3-PD, as a target product, migrated through the membrane; meanwhile, other broth components were rejected in various degrees. Moreover, it was proven that retention of carboxylic acids tended to increase with increasing molecular weight, according to the following order: succinic acid > lactic acid > acetic acid > formic acid. With regards to ions, retention degree increased with the increase of ionic radius and decrease of diffusion coefficient. Finally, it was demonstrated that the CA membrane is resistant to irreversible fouling, which has a positive effect on the economic viability of the process.