Single time point microsomal stability was determined in a 96-well HTS format. Sample preparation was automated using Tecan EVO 200 robot. High Resolution LC/MS (Thermo QExactive) instrument was used to measure the percentage of compound remaining after incubation using a previously described method. Six standard controls were tested in each run: buspirone and propranolol (for short half-life), loperamide and diclofenac (for short to medium half-life), and carbamazepine and antipyrine (for long half-life). 10 mM DMSO stock solutions of the drugs were first diluted to 10 M in 1:2 MeCN:DI H2O and then further diluted to 1 M in assay buffer. Briefly, the incubation consisted of 0.5 mg/mL microsomal protein, 1.0 μM drug concentration, and NADPH regeneration system (containing 0.650 mM NADP+, 1.65 mM glucose 6-phosphate, 1.65 mM MgCl2, and 0.2 unit/mL G6PDH) in 100 mM phosphate buffer at pH 7.4. The incubation was carried out at 37 °C for 15 min. The reaction was quenched by adding 555 μL of acetonitrile (∼1:2 ratio) containing 0.28 μM albendazole (internal standard). Sample acquisition and data analysis was done using a previously described method. 21