Separation, Purification, and Characterization of Diglycerol Linoleic Acid Esters
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Abstract:
Diglycerol linoleic acid esters synthesized by Lipozyme 435 were qualitatively analyzed by thin layer chromatography (TLC), and then separated and purified by silica gel column chromatography. Finally, the purified products were analyzed by high-performance liquid chromatography with an evaporative light scattering detector (HPLC-ELSD) and electrospray ionization mass spectrometry (ESI-MS). The optimal TLC conditions for the separation of diglycerol linoleic acid ester were as follows: sample volume of 3 μL, separation time by chloroform-acetone-methanol solution (96/4/2, V/V/V) of 15 min, coloring time by iodine vapor of 3 min. The optimal silica gel column chromatography conditions for the separation of diglycerol linoleic acid ester were as follows: 2.0 g of sample was dissolved in 5 mL of chloroform-acetone solution (96/4, V/V), separated by a silica gel column (2.8 cm×60 cm, 200~300 mesh) using different mixtures of chloroform-acetone-methanol as the eluent (96/4/0, 95/3/2, 96/4/5, V/V/V) at a flow rate of 1.0 mL/min. Every 10 mL of eluted solution was collected in a tube. The obtained esters of diglycerol linoleic acid were analyzed by differential scanning calorimetry (DSC) and their hydrophilic-lipophilic balance (HLB) values were determined. The results showed that with increasing degrees of esterification, the phase transition temperatures of diglycerol linoleic acid esters increased, while their HLB values decreased.
