Preparation of tritiated alpha-melanotropin with high specific radioactivity

alpha-Melanotropin (alpha-MSH) was iodinated and 3,5-diiodotyr2-alpha-MSH was isolated by reverse phase high performance liquid chromatography (HPLC). Catalytic dehalogenation of the diiodo derivative in the presence of tritium resulted in the formation of 3,5-ditritiotyr2-alpha-MSH. The tritiated peptide was purified by ion exchange and partition chromatography. The radioactive peptide was found to be homogeneous and identical to alpha-MSH by paper electrophoresis and HPLC. The tritiated alpha-MSH stimulated lipolysis in rabbit adipocytes nearly as well as alpha-MSH. The specific radioactivity of tritiated alpha-MSH was 42 Ci/mmol or 73% of the theoretical value.