Determination of the content of metabolites of polyaromatic hydrocarbons in urine by chromatography with mass-selective detection

Introduction. Polyromatic hydrocarbons (PAHs) are metabolized into hydroxyl derivatives, which are excreted from the human body in urine. Gas chromatography with mass-selective (GC-MS) detection is a more sensitive and efficient method for separation and subsequent measurement of hydroxyl PAHs.

Materials and methods. The study used a gas chromatograph with a mass-selective detector and a capillary column HP-5MS. Extraction was performed using dispersive liquid-liquid microextraction, where chloroform served as the extracting solvent, and ethanol as the dispersing solvent. Derivatization of hydroxyl PAHs was carried out online – simultaneously introducing chloroform extract and silylating reagent into the chromatograph injector.

Results. There were calculated characteristics of trimethylsilanes hydroxyl PAH separation on a capillary column as follows: efficiency, separation factor, resolution. The effect of the evaporation port temperature on the detector signal was studied. A dispersing solvent (ethanol, acetone, acetonitrile, methanol) was selected to study its effect on the degree of extraction. The maximum extraction degree is obtained when using ethanol. The patterns of the change in the extraction degree from the ethanol volume and shaking time were studied. The following metrological characteristics were estimated: repeatability, reproducibility, recovery, and accuracy.

Limitations. Improvement of the methodological approach to measuring hydroxyl PAHs in urine was carried out without testing the method on real samples of workers exposed to toxicants in industrial conditions.

Conclusion. In the improved methodical approach of GC-MS determination of hydroxyl PAHs, the use of dispersive liquid-liquid microextraction instead of classical liquid extraction accelerated sample preparation and eliminated the extract evaporation stage. The degree of extraction was 70–100%. Derivatization with the silyl-991 reagent in the evaporation port ensured the rapidity of the reaction.

Compliance with ethical standards. The study does not require submission of the Biomedical ethics committee or other documents.

Contribution:
Alekseenko A.N. – concept and design of the study, search for literature sources, data processing, writing the text;
Zhurba O.M. – collection and processing of material, laboratory research;
Merinov A.V. – collection of literature data, statistical processing, writing the text;
Shayakhmetov S.F. – organization of the study, editing, discussion of results.
All co-authors – approval of the final version of the article, responsibility for the integrity of all parts of the article.

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

Funding. The work was performed within the framework of funds allocated for the implementation of the state task East-Siberian Institute of Medical and Ecological Research.

Received: April 25, 2025 / Revised: August 13, 2025 / Accepted: September 19, 2025 / Published: October 20, 2025

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