Abstract
Human pituitary thyroid-stimulating hormone (TSH) has been analysed by reversed-phase high-performance liquid chromatography on Aquapore RP-300, a 300-Å pore-size short-alkyl-chain silica packing, in sodium chloride-containing buffers. At pH 2.0, two clearly resolved peaks were observed corresponding in retention time to α and β subunit of TSH. With increasing pH, the resolution between these peaks was lost. At pH 7.0 a single major peak was observed, co-eluting with immunoreactive TSH, which could be resolved from β-subunit. In vivo TSH bioactivity was recovered at pH 7.0 in 58% yield. High-performance liquid chromatography of a sample of hypothyroid serum yielded a major peak of TSH immunoreactivity corresponding in retention time to that observed for purified TSH.
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