Reducing agents are crucial for the management of maladaptive inflammation-induced macrophage death and he-matopoietic toxicity of chemotherapy. 2-O-β-D-glucopyranosyl-L-ascorbic acid (AA-2βG), a unique AA (or vitamin C)derivative identified inLyciumbarbarum, exhibited enhanced free radical scavenging activity compared with AA and itssynthetic derivative AA-2αG. AA-2βG protected hydrogen peroxide-induced cell death in murine macrophageRAW264.7 cells. Treatment with AA-2βG eliminated oxidative stress and the ratio of cellular glutathione to glutathionedisulfide more effectively than AA and AA-2αG. AA-2βG also significantly reduced the fluorescent intensity of DCFH-DA triggered by chemotherapeutic agent camptotehcin-11 but not fluorouracil. AA, AA-2αG, and AA-2βG significantlydecreased Keap-1expression, and increased the expression levels of nuclear factor E2-related factor 2 (Nrf2) and hemeoxygenase-1. All compounds triggered the nuclear translocation of Nrf2, while the ability of AA-2βG to enhance theNrf2-DNA binding affinity was approximately two fold as those of AA and AA-2αG. Sodium ascorbate cotransporters(SVCT) inhibitors, sulfinpyrazone, phloretin, and 3-O-methyglucose, potently abrogated the free radical scavengingactivities of AA, AA-2αG, and AA-2βG. The cellular uptake efficacy of AA-2αG and AA-2βG was less than 10% of AA,while the inhibition of SVCT with sulfinpyrazone considerably diminished the uptake efficacy of these compounds. AA-2αG and AA-2βG are more stable in the Fenton reagents than AA. In summary, AA-2βG fromL.barbarumwith excellentfree radical scavenging activity is a promising natural AA derivative for further pharmacological evaluation.