Reliable and rapid analytical methods are the backbone for generating data and deciphering gene functions in the post-genomics era. We describe here a high-throughput method for the rapid profiling of fourteen elements in the 5153 strain gene deletion collection of Saccharomyces cerevisiae. Samples were grown and processed in standard 96-well plate format followed by inductively coupled plasma mass spectrometry (ICP-MS) analysis. Optical densities of the yeast were measured prior to ICP-MS analysis and used for the normalization of the data. The elemental profiling data are stored in an online database for later bioinformatics analysis. This method has the capacity to run 288 yeast samples per day on a single ICP-MS, and has allowed the quantification of the ionome in four replicate cultures of approximately 240 yeast deletion strains per week, along with appropriate wild-type and positive control strains. We identified 400 strains that were outliers from the overall deletion collection in at least one element out of the fourteen that were monitored.