Slit-Mura defect is a notorious yield flaw of color filters. In this study, an innovative non-contact in-line optical inspection method is developed to detect low contrast slit Mura through quantitative measurements by a spectrometer. Using the features of either thickness or chromaticity profiles across a slit Mura, a thickness difference from 21 nm to 41 nm of color filters can be differentiated accurately. Thus, the quality of color filters can be accessed in-line during the manufacturing process TFT-LCDs.