An apparatus and methods for acquiring analytes from a dried biological fluid sample are provided. The apparatus for acquiring the analytes from the dried biological fluid sample includes a tube, a substrate in a proximal section of the tube, and a sorbent bed in a distal section of the tube. A biological fluid sample is dispensed on the substrate and is dried to form a dried sample. A conditioning solvent is flowed into the distal section to condition the sorbent bed. A first elution solvent is flowed through the substrate and the sorbent bed. Analytes are eluted from the dried sample and retained on the sorbent bed. A second elution solvent is flowed through the substrate and the sorbent bed. The analytes are eluted from the sorbent bed, pass through an opening, and are collected. Alternatively, an elution solvent is flowed through the substrate and the sorbent bed such that analytes eluted from the dried sample pass through the sorbent bed and the opening for collection, while non-analytes are retained on the sorbent bed.