Separation of tripalmitin from its hydrolysis products by simple isocratic reversed‐phase high‐performance liquid chromatography

An isocratic reversed‐phase high‐performance liquid‐chromatographic method was developed for monitoring the lipase‐catalyzed hydrolysis of solid triglycerides in supercritical carbon dioxide. The reaction products obtained consist of free fatty acids, monoglycerides, diglycerides, and unreacted triglycerides. For the method, developed with tripalmitin, a mobile phase of acetonitrile/chloroform/acetic acid (50:50:1, vol/vol/vol), a C18 column, and refractive index detection were used. Analysis time is 7 min. Response factors were determined for each neutral lipid class, which permitted quantitation of the hydrolysis product mixture.