Randomly deposited or uniaxially aligned electrospun poly(DL-lactic acid) fibers were prepared, and the molecular orientation was characterized by polarized Fourier-transform infrared (FT-IR) spectroscopy. For randomly deposited fibers, the parallel polarized FT-IR spectrum was the same as that of the perpendicularly polarized spectrum, indicating that there was no specific molecular orientation in this electrospun film. In contrast, for the uniaxially aligned sample, IR band intensities and the positions of C-O-C stretching changed when the IR beam was polarized either parallel or perpendicular to the fiber axis. Because the C-O-C stretching is parallel to the polymer backbone, this change indicates that the polymer chain in the fiber was stretched and aligned. The dichroic ratio of the band at 1090 cm −1 , attributed C-O-C stretching, was 2.9. Furthermore, the high-frequency shift of the peaks in its perpendicularly IR spectra suggests that the polymer chains in the fiber were tightly packed during the electrospinning process.