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Because of their high surface-to-volume ratio, easy surface modification, and simple synthesis methods, gold nanoparticles (AuNPs) are becoming an attractive material as an alternative to conventional solvent extraction and solid-phase extraction column for the selective extraction and enrichment of target analytes from a large sample volume. Through covalent bond formation (Au-S bonds), electrostatic attraction, hydrophobic adsorption, and molecular recognition, AuNPs have been applied successfully to the extraction/removal of a variety of compounds from biological fluids and environment waters, including thiol-containing compounds, peptides, proteins, heavy metal ions, polycyclic aromatic hydrocarbons, and melamine. This review summarizes the recent advances in properties, synthesis, and surface modification of AuNPs for the preconcentration of biomolecules and environmental pollutants.