N-Hydroxysuccinimide


N-Hydroxysuccinimide is an organic compound with the formula 2NOH. It is a white solid that is used as a reagent for preparing active esters in peptide synthesis. It can be synthesized by heating succinic anhydride with hydroxylamine or hydroxylamine hydrochloride.

Activating reagent

NHS is commonly found in organic chemistry or biochemistry where it is used as an activating reagent for carboxylic acids. Activated acids can react with amines to form amides for example, whereas a normal carboxylic acid would just form a salt with an amine.

Use

A common way to synthesize an NHS-activated acid is to mix NHS with the desired carboxylic acid and a small amount of an organic base in an anhydrous solvent. A coupling reagent such as dicyclohexylcarbodiimide or ethyl carbodiimide is then added to form a highly reactive activated acid intermediate. NHS reacts to form a less labile activated acid. The group itself is usually written as SuO- or -OSu in chemical notation.
Such an ester with an acid and NHS, sometimes called succinate ester, is stable enough to be purified and stored at low temperatures in the absence of water and, as such, is commercially available. NHS esters are commonly used for protein modification.

Alternatives

Some alternatives to NHS are the water-soluble analog sulfo-NHS, hydroxybenzotriazole, 1-hydroxy-7-azabenzotriazole, and pentafluorophenol.