Performance of microfiltration (MF) membrane (0.15 µm) in separating total polyphenol from beetroot (Beta vulgaris L.) fermented by Kombucha culture has been a starting reference on application of optimization in semi pilot scale. Separation process was conducted through microfiltration (MF) membrane (pore size 0.15 µm) fitted in Dead-End Stirred Filtration Cell (DESFC) at room temperature, stirrer rotation speed (SRS) of 200, 300 and 400 rpm, and transmembrane pressure (TMP) of 20 and 40 psia for 30 minutes. The experiment result showed that the best optimization of performance of MF membrane fitted in DESFC in separating valuable compounds from beetroot was achieved at SRS of 400 rpm and TMP of 40 psia with flux value of 0.0615 mL/cm2.min. In this process condition was generated retentate (concentrate) and permeate with composition of total polyfenol of 0.67 and 0.43 %, total solids of 10.28 and 9.82%, particles size of 2788.0 and 2570.4 at particle index (PI) of 1.523 and 0.530, respectively. Retentate (concentrate) and permeate were dominated by 2 galic acid monomers and 1 galic acid monomer, molecular weight (MW) of 171.14 and 171.42, and 171.89 Dalton (Da.) (M+) and relative intensity of 100 %, respectively. MF membrane (0.15 µm) fitted in DESFC at this optimum condition is technically able to retain total polyfenols of 123% (1.23 folds), and total solids of 2.08%, compared to prior to process (feed).

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