Polyphenolic antioxidants may effectively reduce acrylamide contents in processed foods. However, few studies focused on their detoxification effects via estimating the profile change of internal exposure biomarkers. Here we showed the protective effect of a water-soluble flavone-C-glycoside-rich antioxidant from bamboo leaves (AOB-w) against acrylamide-induced toxicity in college students. The participants were randomly assigned to either the AOB-w or control group and served potato chips, corresponding to 12.6 μg per kg·bw of dietary exposure to acrylamide, followed by capsules containing 350 mg AOB-w or equivalent placebo. The kinetics of acrylamide, glycidamide, and mercapturic acid metabolites was profiled, and their hemoglobin adducts were measured. The toxicokinetic study showed that AOB-w promoted the excretion of acrylamide and shortened the distribution but prolonged the excretion of N-acetyl-S-(2-carbamoylethyl)-L-cysteine (AAMA) and N-acetyl-S-(2-carbamoyl-2-hydroxyethyl)-L-cysteine. The intervention with AOB-w reduced the peak concentration and area under curve of AAMA by 42.1% and 49.8%, respectively. Besides, AOB-w gender-dependently altered the toxicokinetic profile and reduced the amount of a human-specific urinary biomarker, N-acetyl-S-(2-carbamoylethyl)-L-cysteine-sulfoxide in women. AOB-w accelerated the metabolism of hemoglobin adducts of acrylamide and glycidamide in blood of women. Compared with the baseline levels on the beginning day, we observed a significant enhancement of hemoglobin adducts on the 10th day after serving them potato chips, showing 54.5% and 20.9% higher levels of the hemoglobin adducts of acrylamide and glycidamide, respectively, which thus indicated a lower level of glycidamide-to-acrylamide ratio in blood of participants. Overall AOB-w could effectively reduce the internal exposure to acrylamide in college students, which provides advanced insights into protective functions of natural antioxidants against in vivo toxicity of chemical contaminants from diet.