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In this work, we developed a facile and low-cost method with sufficient sensitivity for the quantitative determination of gold nanoparticles (GNPs) in aqueous and serum solutions.The Au(Ⅲ) digested from GNPs was reduced to Au(Ⅰ) by thiol group of MBI, then Au(Ⅰ)-thiolate complex was formed, which is determined by absorbance at 300 nm.The interference of serum can be avoided in this method.