Developing an express method for quantification of fatty acids in urine based on chromatography-mass spectrometric analysis

Introduction. Quantitative determination of fatty acids (FA) in biological fluids allows assessing of dietary intake, their role in biochemical processes, physiological functions, such as normal brain and heart function.

Objective. To develop a rapid method for the quantitative determination of fatty acids in urine based on chromatograph mass spectrometry analysis for assessing nutritional status.

Materials and Methods. Preparation of urine samples for chemical analysis of fatty acids included alkaline methylation by treating the biosample with potassium methylate, which enabled rapid and quantitative methylation of both free fatty acids and triglycerides. A capillary column with an FFAP with a programmable heating mode of 70–245 °C was used to separate fatty acid esters. The quadrupole mass spectrometer with electron ionization.

Results. The developed rapid method based on chromatographic mass spectrometry (CMS) allows analysing the fatty acid composition in urine, identifying 34 of their derivatives in a concentration range from 0.004 to 9.975 µg/cm³. The conducted studies revealed the following maximum fatty acid concentrations: eicosatrienoic acid (3.608 µg/cm³) in 100% and tricosanoic acid (0.351 µg/cm³) in 91.2% of healthy individuals.

Limitations. Study limitations may be related to the need to exclude the determination of an incomplete spectrum of fatty acids, such as hydroxy acids.

Conclusion. Metabolomic analysis of urine for FA composition may be a promising direction for assessing nutritional status and identifying biomarkers characterizing changes in alimentary factors.

Compliance with ethical standards. The study was approved by the local ethics committee of the Rospotrebnadzor’s Federal Scientific Center for Medical and Preventive Health Risk Management Technologies (the meeting protocol No. 3 dated March 28, 2025). All participants gave informed voluntary written consent to participate in the study.

Contribution:
Nurislamova T.V., Lear D.N. – concept, study design, editing;
Terentyev G.I. – collection and treatment of material, analysis of obtained data, writing the text;
Subbotina D.Yu., Yakusheva E.A. – collection and treatment of material, analysis of obtained data;
Suvorov D.V. – collection and treatment of material.
All authors are responsible for the integrity of all parts of the manuscript and approval of the manuscript final version.

Conflict of interest. The authors declare no conflict of interest.

Funding. The study had no sponsorship.

Received: September 9, 2025 / Revised: October 24, 2025 / Accepted: November 3, 2025 / Published: December 19, 2025

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