(A) The high-performance liquid chromatographic with a diode array detection, (HPLC-DAD) profile of human thrombin purified using a reverse-phase column (Discovery® BioWide Pore C18, 25 cm × 10 mm, 10 μm). The sample was eluted with buffer A (0.1% trifluoroacetic acid (TFA)) and buffer B (66% acetonitrile (ACN) and 0.1% TFA) at a flow rate of 2 mL/min and the following gradient: 5 min, 100% buffer A; 30 min, 100% buffer B; and 36 min, 100% buffer A; (B) The HPLC profile of purified thrombin measured at 288 nm; (C) UV-Vis spectra of purified thrombin analyzed by performing UV scanning from 190 nm to 500 nm.