As with other derivatization reagents, alkylation reagents reduce molecular polarity by replacing active hydrogens with an alkyl group. These reagents are used to modify compounds having acidic hydrogens, such as carboxylic acids and phenols. Alkylation reagents can be used alone to form esters, ethers and amides or they can be used in conjunction with acylation or silylation reagents. A two-step approach is commonly used in the derivatization of amino acids, where multiple functional groups on these compounds may necessitate protection during derivatization.

Esterification, the reaction of an acid with an alcohol in the presence of a catalyst to form an ester, is the most popular method of alkylation, due to the availability of reagents and ease of use. Alkylation reagents are available in several configurations that enable the formation of a variety of esters. Alkyl esters are stable, and can be formed quickly and quantitatively. By altering the length of the substituted alkyl group, retention of the derivative can be varied. In addition to the formation of simple esters, alkylation reagents can be used in extractive procedures where biological matrices can be present.

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