Practical Protein Chemistry - A. Darbre 1989

Application of electron impact mass spectrometry for determining the amino acid sequence of peptides and proteins
Sample preparation methods
Hydrazinolysis

50 µl of an aqueous hydrazine hydrate solution (1:1) is added to 5–100 nmol of the peptide, and the solution is incubated for 12 min at 80 °C. The reaction mixture is then cooled, 50 µl of Water is added, the solution is frozen using liquid nitrogen or an ethanol-dry ice mixture, and evacuated with a forevacuum pump until moisture is completely removed (a freeze dryer is convenient to use). The sample frozen in this way is slowly thawed and brought to room Temperature. This Procedure is accompanied by the release of gases dissolved in the reaction mixture. Residual hydrazine hydrate is removed by twice adding 50 µl of water to the sample, which is then evaporated in vacuo.



Last update: 06/08/2026

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