In-line determination of the enantiomeric ratio is still a challenge in process analytical technology (PAT). This study combines enantioselective Raman (esR) spectroscopy with partial least-squares regression (PLSR) to determine the enantiomeric fraction of the chiral molecule (5,6)-diphenyl-morpholin-2-one diluted in dimethyl sulfoxide (DMSO) as a proof-of-concept. Morpholinone derivates are potential candidates for pharmaceutical applications. The PLS weights were carefully analyzed in order to avoid misleading regression results, e.g. caused by sample impurities. A suitable PLSR model was found with two components and it was validated by a leave-one-out cross-validation. The enantiomeric fraction ef(+) could be calculated with deviations from the prepared ef(+) in the range of -0.031 and +0.052 from the esR spectra recorded at a half-wave retarder angle of 30.0°.