This research presents a comprehensive study of sequential oxidative extraction (SOE) consisting of alkaline and acidic oxidation processes to extract nanocellulose from plant biomass. This proposed process is advantageous as its operation requires a minimum process with mild solvents, and yet successfully isolated high-quality nanofibrillated cellulose (NFC) from raw OPEFB. The SOE involved ammonium hydroxide (NH4OH, 2.6 M) and formic acid (HCOOH, 5.3 M) catalyzed by hydrogen peroxide (H2O2, 3.2 M). This approach was used to efficiently solubilize the lignin and hemicellulose from Oil Palm Empty Fruit Bunch (OPEFB) at the temperature of 100°C and 1 h extraction time, which managed to retain fibrous NFC. The extracted solid and liquor at each stage were studied extensively through physiochemical analysis. The finding indicated that approximately 75.3%dwb of hemicellulose, 68.9%dwb of lignin, and 42.0%dwb of extractive were solubilized in the first SOE cycle, while the second SOE cycle resulted in 92.3%dwb, 99.6%dwb and 99.8%dwb of solubilized hemicellulose, lignin, and extractive/ash, respectively. High-quality NFC (75.52%dwb) was obtained for the final extracted solid with 76.4% crystallinity, which is near the crystallinity of standard commercial NFC. The proposed process possesses an effective synergy in producing NFC from raw OPEFB with less cellulose degradation, and most of the degraded hemicellulose and lignin are solubilized in the liquor.