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Abstract: Recent advances in analytical chemistry, such as miniaturization, automation, robotics, parallel separations, detection systems and chemometrics, have found interesting applications in the development of drug-discovery programs. To select drug candidates, these programs require high-throughput screening (HTS) assays, which evaluate the biological activity of a large number of compounds against a panel of targets, as well as determining the physicochemical and pharmacological properties of these compounds. However, the need to obtain such information earlier in the drug-discovery process has posed new challenges to analytical chemistry. This article presents a critical overview of: [•] the current analytical techniques and approaches used in selecting drug candidates; [•] the limitations that need to be overcome; and, [•] the recent advances in HTS assays proposed to overcome these limitations. [Copyright &y& Elsevier]