Method

Tryptophan oxidation during hydrolysis is prevented by using thioglycollic acid (TGA) as the reducing acid.

Hydrolysis solution. 7 M hydrochloric acid containing 1 per cent of phenol, 10 per cent of trifluoroacetic acid and 20 per cent of thioglycollic acid.

Vapour phase hydrolysis. Dry about 10 pg to 50 pg of the protein/peptide under test in a sample tube. Place the sample tube in a larger tube with about 200 pl of the hydrolysis solution. Seal the larger tube in vacuo (about 50 pm of mercury or 6.7 Pa) to vaporise the TGA. Heat the sample tube to 166 °C for about 15-30 min. After hydrolysis, dry the sample tube in vacuo for 5 min to remove the residual acid. Recovery of tryptophan by this method may be dependent on the amount of sample present.

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