Costs and resource efficiency of laser powder bed fusion (L-PBF) are highly dependent on the ability to produce high quality parts with recycled powders. There is a need to control the quality of the material, which has a direct influence on the performance of the printed parts. Particles oxidation is known to increase with repeated powder recycling and can be a good indicator of powder degradation. The characterization of powders oxygen content is time-consuming, expensive, and usually carried out ex-situ on non-reusable quantities that are not necessarily representative of the entire feedstock. In this work, a new methodology was developed to measure the oxygen content of powders by in-line scanning of powder bed layers. The method takes advantage of stainless steel particles coloration related to their oxidation level in order to assess their oxygen concentration as a function of Red, Green and Blue channel values of image scans. The calibration procedure once carried out, several recycled powder samples were scanned and analyzed, and the determined powder beds oxygen contents were demonstrated to be in accordance with ex-situ measurements. The results highlight a new opportunity to monitor and evaluate powder degradation in-situ on powder bed layers by image analysis.