Growth in the international popularity of Brazil nuts oil (Bertholletia excelsa, Bonpl.) has demanded a closer look at quality aspects of the nuts. In this context, time-domain 1H nuclear magnetic resonance spectroscopy (TD-NMR) can be a useful tool for assessing the lipid portion of such matrices. In this work, a TD-NMR method was developed and validated to assess the oil content directly from Brazil nuts following the ISO 10565:1998 and DOQ-CGCRE-008. Full validation was addressed covering the working range from 0.67 to 25.00 g (coefficient of determination, R2= 0.9993). Limits of detection (LOD) and quantification (LOQ) were 0.20 and 0.67 g, respectively. The repeatability and intermediate precision assays met the guide requirements. Oil contents obtained in two laboratories (different equipment) were statistically similar (66.93 ± 4.69 and 70.13 ± 4.15%), highlighting the method’s robustness. The developed approach has the potential to be adopted as an industrial reference method for in situ monitoring of Brazil nuts oil contents.