Direct determination by high-performance liquid chromatography of sn-2 monopalmitin after enzymatic lipase hydrolysis

An alternative method to determine the sn-2 monopalmitin in infant formulas was developed and validated. This method offers many advantages over the traditional methods. It follows the official method in the first steps, purification of the fat or oil through an alumina column, and subsequently the triglycerides are incubated with pancreatic lipase in order to obtain the sn-2 monoglycerides. In traditional methods the sn-2 monoglycerides are separated by preparative thin-layer chromatography and then, the 2-monoglycerides are converted into the corresponding fatty acid methyl esters and analysed by gas chromatography. In our method, separation, quantification and identification of the sn-2 monoglycerides were achieved by high-performance liquid chromatography with evaporative light-scattering detection. The detection limit (0.19 microg), quantification limit (0.38 microg), linearity range (r=0.999, range 1-200 microg) and precision (SD=1.10) show the suitability of the proposed method. This method is faster, cheaper and simple and does not consume large quantities of reagents and materials.