New, rapid method to measure dissolved silver concentration in silver nanoparticle suspensions by aggregation combined with centrifugation

It is unclear whether the antimicrobial activities of silver nanoparticles (AgNPs) are exclusively mediated by the release of silver ions (Ag(+)) or, instead, are due to combined nanoparticle and silver ion effects. Therefore, it is essential to quantify dissolved Ag in nanosilver suspensions for investigations of nanoparticle toxicity. We developed a method to measure dissolved Ag in Ag(+)/AgNPs mixtures by combining aggregation of AgNPs with centrifugation. We also describe the reproducible synthesis of stable, uncoated AgNPs. Uncoated AgNPs were quickly aggregated by 2 mM Ca(2+), forming large clusters that could be sedimented in a low-speed centrifuge. At 20,100g, the sedimentation time of AgNPs was markedly reduced to 30 min due to Ca(2+)-mediated aggregation, confirmed by the measurements of Ag content in supernatants with graphite furnace atomic absorption spectrometry. No AgNPs were detected in the supernatant by UV-Vis absorption spectra after centrifuging the aggregates. Our approach provides a convenient and inexpensive way to separate dissolved Ag from AgNPs, avoiding long ultracentrifugation times or Ag(+) adsorption to ultrafiltration membranes.