Poly(lactic acid)/hexadecyl trimethyl ammonium chloride-modified montmorillonite clay nanofiber membranes have been fabricated by the electrospinning technique. The nanofiber membranes were then characterized by scanning electron microscopy and Fourier transform-infrared spectroscopy. Nanofiber membranes with different ratios of Poly(lactic acid) and hexadecyl trimethyl ammonium chloride-modified montmorillonite have been spun by varying voltage supply during electrospinning. These parameters were found to have a substantial effect on the morphologies of these membranes. It was found that 8% (w/w) Poly(lactic acid)/hexadecyl trimethyl ammonium chloride-modified montmorillonite concentration is an ideal condition to obtain thinner and uniform Poly(lactic acid) fibers. The results also suggested the coexistence of exfoliated hexadecyl trimethyl ammonium chloride-modified montmorillonite layers over the studied hexadecyl trimethyl ammonium chloride-modified montmorillonite contents. Fourier transform-infrared spectroscopy revealed that there might be possible interactions between the hexadecyl trimethyl ammonium chloride-modified montmorillonite clay and Poly(lactic acid) matrix.