A method for rapid and objective assessment of ocular lens density and transmittance is needed for research and clinical practice. The aim of this study was to determine whether the Purkinje image-based technique can be used for objective and accurate quantification of spectral density and transmittance of ocular media (the mainly crystalline lens) in visible light. Twenty-six individuals (10 young, 9 middle-aged and 7 older individuals) participated in this study. Spectral lens density was evaluated by detecting the intensity of the IVth Purkinje image for different wavelengths. Subsequently, optical density index (ODI), the area under the curve in the lens density spectrum, was calculated and ODIs were compared with clinical lens opacification scales assessed subjectively using a slit lamp. Spectral lens transmittance was estimated from the lens density spectrum. Lens densities were higher in the short wavelength region of the visible spectrum across all age groups. ODI was highly correlated with the clinical opacification scale, while lens transmittance decreased with aging. Our results showed that spectral transmittance of the human crystalline lens can be easily estimated from optical density spectra evaluated objectively and rapidly using the Purkinje image-based technique. Our results provide clinicians and scientists with an accurate, rapid and objective technique for quantification of lens transmittance.