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Simultaneous determination of 22 fatty acids in total parenteral nutrition (TPN) components by gas chromatography-mass spectrometry (GC-MS)
Uppsala University, Disciplinary Domain of Science and Technology, Chemistry, Department of Chemistry - BMC, Analytical Chemistry. Fresenius Kabi, Innovat & Dev Dept, Uppsala, Sweden..ORCID iD: 0000-0001-9682-6840
Uppsala University, Disciplinary Domain of Science and Technology, Chemistry, Department of Chemistry - BMC, Analytical Chemistry.
Fresenius Kabi, Innovat & Dev Dept, Uppsala, Sweden..
Swedish Food Agcy Livsmedelsverket, Uppsala, Sweden..
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2023 (English)In: Analytical Methods, ISSN 1759-9660, E-ISSN 1759-9679, Vol. 15, no 20, p. 2480-2489Article in journal (Refereed) Published
Abstract [en]

Evaluating total parenteral nutrition (TPN) products for quality assurance and quality control is crucial due to the chemical complexity of its components. With the advent of exploring different approaches for analysing TPN components using tandem mass spectrometry techniques, there is still a need for a robust and reproducible method for industrial routine analyses. This study allows simple, simultaneous determination of 22 fatty acids (FAs) commonly found in TPN components using gas chromatography-mass spectrometry (GC-MS). Five different transesterification techniques were applied for the FA standards and the sodium methoxide in methanol-dimethyl carbonate method was selected due to its good methylation efficiency. Fatty acid methyl esters (FAMEs) were separated in gas chromatography using an HP-5MS UI column with helium as the carrier gas. Mass spectrometry was used to fragment and quantify FAMEs using electron ionization (EI) and selected ion monitoring (SIM) mode. The analytical method was evaluated using the guidelines from the US Food and Drug Agency (FDA) and European Medicines Agency (EMA) in compliance with the International Council for Harmonization (ICH) document Q2(R2). Correlation coefficients (R-2) of the calibration curves for FAMEs were 0.99, except for C24:1 n-9 and C24:0, both R-2 = 0.98. The limits of detection (LOD) and quantification (LOQ) were found to be 1.69 mu g mL(-1) and 5.14 mu g mL(-1), respectively. The linear range was from 3.10-179.9 mu g mL(-1) for most FAMEs, except for C18:1 n-7 (3.96-224.9 mu g mL(-1)) and C18:1 n-9 (6.30-349.57 mu g mL(-1)). The intra-day and inter-day precision coefficients of variance (CV) of the method were less than 11.10% and 11.30%, respectively. Freeze-thaw cycles and ambient temperature measurements were performed for assessing sample stability. The validated method was applied to analyse major TPN components-fish and olive oils, and an unidentified lipid sample. The presented GC-MS method is simple and robust in the identification and quantification of 22 fatty acids simultaneously in the tested TPN components.

Place, publisher, year, edition, pages
Royal Society of Chemistry, 2023. Vol. 15, no 20, p. 2480-2489
National Category
Analytical Chemistry
Identifiers
URN: urn:nbn:se:uu:diva-504041DOI: 10.1039/d3ay00407dISI: 000987653200001PubMedID: 37183597OAI: oai:DiVA.org:uu-504041DiVA, id: diva2:1776392
Available from: 2023-06-28 Created: 2023-06-28 Last updated: 2025-02-02Bibliographically approved
In thesis
1. Development and validation of chromatography and mass spectrometry-based lipidomic methods for pharmaceutical lipid emulsion components in total parenteral nutrition
Open this publication in new window or tab >>Development and validation of chromatography and mass spectrometry-based lipidomic methods for pharmaceutical lipid emulsion components in total parenteral nutrition
2025 (English)Doctoral thesis, comprehensive summary (Other academic)
Abstract [en]

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.

Place, publisher, year, edition, pages
Uppsala: Acta Universitatis Upsaliensis, 2025. p. 94
Series
Digital Comprehensive Summaries of Uppsala Dissertations from the Faculty of Science and Technology, ISSN 1651-6214 ; 2497
Keywords
Chromatography, mass spectrometry, targeted lipidomics, method development, method validation, pharmaceutical lipid emulsion, total parenteral nutrition
National Category
Analytical Chemistry
Research subject
Chemistry with specialization in Analytical Chemistry
Identifiers
urn:nbn:se:uu:diva-549282 (URN)978-91-513-2370-1 (ISBN)
Public defence
2025-03-14, room A1:107a, Biomedical Centre (BMC), Husargatan 3, Uppsala, 09:15 (English)
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Supervisors
Available from: 2025-02-19 Created: 2025-02-02 Last updated: 2025-02-19

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Retrato, Mark Dennis ChicoRad, Farshid MashayekhyUbhayasekera, KumariBergquist, Jonas

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