An Improved Liquid Chromatographic Method for the Quantitative Determination of Free Fatty Acids in Milk Products

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Abstract
An existing HPLC method for the quantitative determination of the major FFA in milk fat was improved. Complete resolution of the p-bromophenacyl (PBP) ester derivatives of saturated and unsaturated FFA of acyl-chain length C4 to C18 was accomplished by gradient elution (60 to 100% acetonitrile in water) when the temperature of the reverse-phase column was maintained at 10.degree. C. This allowed the determination of all the major FFA in milk fat within a single chromatographic analysis. The improved method should save time in analysis of FFA in milk fat and also can be used to quantify the low concentrations (78 to 270 .mu.M .+-. 17%) of the major FFA (myristic, palmitic, stearic, and oleic acids) in fresh pasteurized milk and follow the time course of milk fat hydrolysis where low amounts of FFA are liberated.