We report the first case in the literature of fatal betaxolol (Kerloneρ) self-poisoning. After a single-step liquid-liquid alkaline extraction, betaxolol was identified by a high-performance liquid chromatographic-diode array detection screening procedure and then quantitated in cardiac blood, heart, brain, muscle, spleen, stomach contents, duodenum contents, liver, and kidney. The blood betaxolol concentration was 36 mg/L.