Phospholipid profiling : a computationally assisted LC-HRMS approach in lecithin

Šijanec, Ana (author), Pompe, Matevž (author)

The use of lecithin as an emulsifier in food supplements has increased in recent years. However, successful formation of liposomes or micelles requires an appropriate mixture of phospholipids in lecithin. To evaluate the emulsification properties of lecithin for food supplements, a reliable analytical procedure for characterizing phospholipids is necessary. A liquid chromatography–mass spectrometry method was developed to identify phospholipids in lecithin without standard reference materials. For efficient separation of phospholipids before mass spectrometric analysis, a reverse-phase high-performance liquid chromatography method was optimized using a Waters XBridge Protein BEH C4 column. The optimized chromatographic method demonstrated good linearity and precision. Molecular ions were detected in full scan mode to determine accurate mass-to-charge ratios for individual peaks in the chromatogram. A custom Python program was then used to generate a list of possible phospholipid species for each peak based on the measured mass-to-charge ratios. Tandem mass spectrometry was performed to confirm the identity of specific phospholipids by comparing experimental fragmentation patterns with theoretical predictions. Identification of the phospholipids was also confirmed with four commercially available standard reference compounds, demonstrating the reliability of the proposed approach. The developed method offers a practical and cost-effective strategy for identifying phospholipids in complex matrices, especially when standard reference compounds are unavailable. Additionally, it enables targeted selection of standard compounds for future quantitative analyses, making it a valuable tool for comprehensive lipid profiling.

• Language: English
• Keywords: phospholipids, lecithin, liquid chromatography, mass spectrometry
• Work type: Article
• Typology: 1.01 - Original Scientific Article
• Organization: FKKT - Faculty of Chemistry and Chemical Technology
• Publication status: Published
• Publication version: Version of Record
• Year: 2026
• Number of pages: 13 str.
• Numbering: Vol. 13, iss. 1, art. 40
• PID: 20.500.12556/RUL-179675
• UDC: 543.544.5:543.51
• ISSN on article: 2297-8739
• DOI: 10.3390/separations13010040
• COBISS.SI-ID: 265853955
• Publication date in RUL: 19.02.2026

Record is a part of a journal

• Title: Separations
• Shortened title: Separations
• Publisher: MDPI AG
• ISSN: 2297-8739
• COBISS.SI-ID: 525934873

Licences

• License: CC BY 4.0, Creative Commons Attribution 4.0 International
• Link: http://creativecommons.org/licenses/by/4.0/
• Description: This is the standard Creative Commons license that gives others maximum freedom to do what they want with the work as long as they credit the author.

Secondary language

• Language: Slovenian
• Keywords: fosfolipidi, lecitin, tekočinska kromatografija, masna spektrometrija

Projects

• Funder: ARIS - Slovenian Research and Innovation Agency
• Project number: P1-0153-2020
• Name: Raziskave in razvoj analiznih metod in postopkov