Mark Dennis Chico Retrato: Development and validation of chromatography and mass spectrometry-based lipidomic methods for pharmaceutical lipid emulsion components in total parenteral nutrition

Abstract

Total parenteral nutrition (TPN) is a life-sustaining therapy that delivers essential nutrients intravenously to patients unable to meet their dietary requirements through oral intake. TPN formulations typically contain a mixture of carbohydrates, proteins, lipids, vitamins, and minerals, with pharmaceutical lipid emulsions (PLEs) serving as a key component. Ensuring the stability and quality of TPN lipids is critical as compositional changes—particularly in PLEs, can impact formulation efficacy and patient safety.

This thesis explores the lipidomic analysis of PLEs by investigating lipid stability and degradation over time. This research develops and applies chromatography coupled with mass spectrometry (MS): gas chromatography (GC-MS) for Paper I, supercritical fluid chromatography (SFC-MS) for Papers II-III and liquid chromatography (LC-MS) for Paper IV methods to investigate important lipid groups, including free fatty acids (FFAs), cholesterol and cholesterol oxidation products (COPs), phospholipids (PLs), and triacylglycerols (TAGs). These tailored lipidomic techniques provided critical insights into compositional changes that may indicate PLE degradation and potential TPN instability.

To ensure analytical robustness, all methods were validated according to ICH Q2(R2) guidelines meeting pharmaceutical quality standards. Present study also addresses matrix effects and emphasizes the importance of using appropriate internal standards for accurate lipid quantification. The developed strategies were applied to pharmaceutical-grade egg yolk powders, a key raw material for PLE formulations. These findings contribute to improving lipidomic methodologies for quality control, enabling high-throughput, and reproducible analysis of TPN formulations, supporting safer and more effective patient care.