Fatty acids are major lipid components, and their content ratios exhibit some features of lipids. In this note, utilizing ADAM (9-anthryldiazomethane) as a fluorescent derivatization reagent, fatty acids are determined by HPLC-fluorescence detection system.
In example 1, the derivatives of representative fatty acids, such as EPA (cis-5,8,11,14,17-eicosapentaenoic acid) and DHA (cis-4,7,10,13,16,19-docosahexaenoic acid), are successfully separated. However, run time becomes too long if more hydrophobic fatty acids are analyzed under the condition. Therefore, another condition shown in example 2 was also described in this note.
The derivatization reaction was peroformed by adding 100μL of 0.1 % ADAM solution to 500 μL of standard solution of fatty acids. After waiting for 1 hour, the reaction solution was injected into the HPLC system.