The separation of chiral molecules using membranes has garnered much research interest. In this study, the enantioseparation of amino acids using chiral membranes, namely graphene oxide-ethylenediamine-maltodextrin (GO-EDA-MD) and GO-EDA-hydroxypropyl-MD (GO-EDA-HP-MD), was evaluated. HP-MD and MD were investigated as chiral selectors due to their inherent chirality. Various characterization techniques, including atomic force microscopy, Fourier transform infrared spectrometry, field emission scanning electron microscopy, water contact angle analysis, tensile properties, and thermal gravimetric analysis were employed to analyze the membrane structures. The evaluation of enantioseparation performance was conducted by employing tryptophan, phenylalanine, and tyrosine enantiomers. The optimal conditions for separating enantiomers comprised the utilization of the GO-EDA-HP-MD chiral membrane, with concentrations of 1.3 wt% and 0.6 wt% for HP-MD and GO, respectively. The feed concentration was maintained at 10 mg/L, the operation time at 15 min, and the membranes effective surface area at 1.0 cm2. Also, the bovine serum albumin rejection was 90.0%, and the water flux reached 37.1 L.m−2.h−1. The highest enantiomeric excess (ee.%) values were obtained at 46.33%, 76.97%, and 73.04% for tryptophan, phenylalanine, and tyrosine, respectively. The impact of voltage on ee.% and substance flux was also explored. This membrane was able to separate enantiomers successfully.