Hydrochloric Acid → Amphetamine

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Amphetamine-ATP (0.095 g, 3×10-4 mol, Example 12) was dissolved in 50 ml of 0.12M potassium carbonate in 80% methanol/20% water. After 5 min at room temperature the thiol concentration was determined by reaction with DTNB to be 25 mM. The amphetamine thiol (7.6 ml, 1.9×10-4 mol) was added with stirring to 20.3 ml (0.4 g, 5.9×10-6 mol) of SMCC-BSA, 10/1. The pH of the solution was adjusted to 7 with 1N hydrochloric acid. The container was purged with argon and the solution was stirred at room temperature for 2 hr. The conjugate solution was then dialyzed with a PYROSTART ultrafiltration module against 10 mM morpholineethanesulfonic acid, pH 5.77 and 110 ml of 3.57 mg/ml BSA-amphetamine was collected. The protein solution was frozen at -70° C.

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