Ninhydrin Solid Peptide Synthesis
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Ninhydrin also called (Triketohydrindane hydrate) is a chemical used to detect ammonia or primary amines. Ninhydrin is also used to monitor deprotection in solid phase peptide synthesis. Deprotection of the growing peptide chain yields blue with a ninhydrin test. If the next peptide residue is coupled then the test is colorless or yellow.
Ninhydrin Test Protocol
- Aliquot 20μL of resin from the reaction tube into filter tube.
- Wash the resin. Aliquot with DMF and DCM, fast flow. Rinse well.
- Dry aliquot until it is a powder.
- Transfer powder into glass culture tube.
- Add 40μL phenol / ethanol + 40μL KCN / pyridine + 20μL Ninhydrin / ethanol to the resin aliquot.
- Heat the tube at 95 °C for 2-5 minutes.
- Bluish color indicates an unfinished reaction.
- A completed reaction looks pink.