Separation of lipid components on TLC plates is a function of their polarity. Fresh activation of TLC plates by heating at 110 °C in oven for 30 min before applying the samples for separation was essential for removal of impurities in the silica gel that would otherwise interfere with separation of lipid classes.As established by Christie (1993), all fatty acids are es- terified at approximately the same rate by methanolic hydrogen chloride; therefore, there is minimum loss of specific fatty acids during the esterification process. However, additional precautions were taken during es- terification step such as avoiding high temperature for prolonged periods of time and vigorous agitations so as to prevent degradation and loss of short-chain esters. Compounds that may interfere during GC analysis may be formed from the methanol if prolonged heating at high temperature is allowed in the presence of oxygen. Samples containing PUFA requires handling with care and not to be subjected to vigorous conditions like pro- longed heating, vortexing and agitation. Reagents that are perfectly satisfactory when used under optimized conditions can be destructive to fatty acids if used care- lessly (Christie 1993).The primary objective of the validation process for the fatty acids analysis was to establish the basis for a writ- ten procedure for process control and to demonstrate that the process is suitable for the intended purpose. A summary of the validation parameters undertaken is pre- sented here.