Potassium Hydroxide → Morphine

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Morphine-HCTL (0.068 g, 1.3×10-4 mol, example 9) was dissolved in 5.7 ml of 70% dimethylformamide/30% water and 1.43 ml 1N potassium hydroxide was added. After 5 min the thiol concentration was determined by reaction with DTNB to be 16.9 mM. The morphine thiol -- (6.4 ml, 1.1×10-4 mol) was added with stirring to 26.7 ml SMCC-BSA, 15/1, (0.16 g, 2.4×10-6 mol). The container was purged with argon and the solution was stirred at room temperature for 2 hr. The protein solution was then dialyzed using a PYROSART module against 10 mM (2-(N-morpholino) ethane sulfonic acid, pH 5.77, and 29.5 ml of 5.39 mg/ml BSA-morphine was collected. The protein solution was frozen at -70° C.

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