Characterization of pharmaceutic structured triacylglycerols by high-performance liquid chromatography/tandem high-resolution mass spectrometry and its application to structured fat emulsion injection

Rationale: Structured triacylglycerols (STAGs) are a complex mixture of triacylglycerols which are esterified by long-chain fatty acids and medium-chain fatty acids with the same glycerol molecular backbone. As a kind of lipid pharmaceutic excipients, STAGs are used in the pharmaceutical industry as major components of structured fat emulsion injections and play an important role in pharmaceutic energy material because they improve nutritional status with faster elimination in a safe way. The composition and proportion of triacylglycerols in STAGs are closely related to the stability of pharmaceutical preparations and curative effects in the clinic. Therefore, it is necessary to characterize pharmaceutic STAGs using a rapid and accurate method.

Methods: An analytical method for rapid and accurate determination of triacylglycerols in pharmaceutic STAGs was developed using high-performance liquid chromatography/tandem high-resolution mass spectrometry (HPLC/HRMS). Triacylglycerol components could be well separated on a Waters Xterra MS C8 (2.1 × 100 mm, 3.5 μm) column. Four-dimensional HPLC/HRMS data (high-resolution m/z, MS² data, retention time and isotopic intensity distribution) were used to identify triacylglycerols using Lipid Data Analyzer (LDA) software and the LIPID MAPS database. Then, these identified triacylglycerol components were relatively quantified by their corresponding normalized peak areas using representative standard curves of structurally similar standard substances.

Results: Forty-seven kinds of triacylglycerol components in pharmaceutic STAGs and structured fat emulsion injection were identified and relatively quantified by this method. It has been shown that their retention times are in good correlation with the number of carbon atoms and carbon-carbon double bonds. The main components in pharmaceutic STAGs and structured fat emulsion injection were triacylglycerols containing both medium-chain fatty acids and long-chain fatty acids, while the other components, including triacylglycerols containing three medium-chain fatty acids and triacylglycerols containing three long-chain fatty acids, were relatively low.

Conclusions: This study has provided a rapid and accurate approach for the identification and quality control of pharmaceutic STAGs and structured fat emulsion injection and this approach can be extended to other lipid pharmaceutic excipients and used as an effective and reasonable control to guarantee the quality of pharmaceutical preparations.