A Comparison of Sodium Hydroxide and Sodium Sulfide Digestion of Mouse Hair in the Recovery of Radioactivity Following Systemic Administration of [3H]-Nicotine and [3H]-Flunitrazepam

Pigmented (C57Bl) and nonpigmented (balb/c) mice, 25 days of age, were treated intraperitoneally with [3H]-nicotine (4 mg/kg, 555 dpm/ng) or [3H]-flunitrazepam (1 mg/kg, 2200 dpm/ng) daily for three days. After 21 days, shaved back hair was digested at 37°C for 24 h with either 1M sodium hydroxide or 1M sodium sulfide. With both drugs, sodium sulfide extraction removed the same amount of radioactivity as sodium hydroxide from nonpigmented hair. However, sodium sulfide removed significantly more radioactivity from pigmented hair than did sodium hydroxide. In pigmented hair, sodium sulfide solubilized 35% and 74% of the flunitrazepam- and nicotine-associated radioactivity, respectively. Of this, 12% and 43%, respectively, could be partitioned into ethyl acetate. Microscopic examination of residual pellets after digestion demonstrated a more thorough dissolution of the hair shaft with sodium sulfide with only melanosomes remaining. The results demonstrate the significant interaction of flunitrazepam and nicotine with melanins and the utility of sodium sulfide in increasing drug recovery.