Normal-phase high performance liquid chromatographic separation and characterization of short- and long-chain triacylglycerols

Abstract

Short-and long-chain triacylglycerols (SLCT) are a family of lipids prepared by chemical or enzymatic interesterification of triacetin, tripropionin and/or tributyrin, and long-chain (C16!18) hydrogenated vegetable oils. In this study, a normal-phase cyanopropyl high-performance liquid chromatographic (HPLC) method was developed for the separation and quantification of SLCT. The method is capable of separating SLCT mixtures, free fatty acids and the neutral lipid classes of saturated long-chain triacylglycerols, diacylglycerols and monoacylglycerols. To characterize the specific SLCT classes, a normal-phase HPLC procedure using a non-modified silica column was developed to separate the SLCT into individual isomers based on total carbon number and position of fatty acids on the glycerol backbone. Online coupling with a mass detector (LC/MS) allowed the identification of the individual triacylglycerol structures.Original 0009-5893/99/04

Separation of structured lipids by high performance liquid chromatography

“…The ratio of the two SL types in the final product was dependent on the molar ratio of reactants used. From previous studies on the interesterification of triacetin with HSO, a ratio of 2:1 was optimal [7] and this was the ratio used in this work. HPLC analysis of the MLCT product produced by acidolysis of tributyrin with stearic acid under the reaction conditions used in this study gave the chromatograms shown in Figure 2.…”

“…The NPcN-HPLC gradient used was reported previously [6]. For the NPslL-HPLC method, an isocratic binary mobile phase consisting of 15% MTBE and 85% hexane (v/v, each fortified with 0.4% acetic acid) was used at a flow rate of 0.75mLmin 1 [7]. The ELSD detector was operated at 40 ~ with nitrogen nebulizing gas flow of 1.51 SLPM for both NP methods.…”

Separation of Lipid Classes by HPLC on a Cyanopropyl Column

“…In the last few years, PVA columns [11,12], diol columns [13,14], and amino columns [15,16] have been used by other groups in the separation of polar lipid classes. Cyanopropyl columns have been employed in the separation of non-polar lipid classes, mainly in applications related to transesterification reaction products [17][18][19].…”

Mapping the regioisomeric distribution of fatty acids in triacylglycerols by hybrid mass spectrometry

“…The applicability of this approach is limited in complex mixtures with abundant interferences, and it does not provide information about the identity of individual TAG species. Several chromatographic methods were implemented for the separation of TAG ( 27 ), among which the three mainstream approaches are normal phase liquid chromatography (NP-LC) (28)(29)(30), silver-ion liquid chromatography (Ag-LC) ( 4,5,27,(31)(32)(33)(34), and nonaqueous reversed phase liquid chromatography (NARP-LC) ( 5,13,19,(35)(36)(37)(38)(39).…”