medlitГигиена и санитарияHygiene and Sanitation0016-99002412-0650Federal Scientific Center of Hygiene named after F.F. Erisman10.47470/0016-9900-2025-104-9-1209-1215lulvjamedlit-5164Research ArticleМЕТОДЫ ГИГИЕНИЧЕСКИХ ИССЛЕДОВАНИЙMETHODS OF HYGIENIC AND EXPERIMENTAL INVESTIGATIONSОпределение содержания метаболитов полиароматических углеводородов в моче методом хроматографии с масс-селективным детектированиемDetermination of the content of metabolites of polyaromatic hydrocarbons in urine by chromatography with mass-selective detectionhttps://orcid.org/0000-0003-4980-5304АлексеенкоАнтон НиколаевичAlekseenkoAnton N.

Канд. хим. наук, ст. науч. сотр. ФГБНУ ВСИМЭИ, 665827, Ангарск, Россия

e-mail: alexeenko85@mail.ru

PhD (Chemistry), senior researcher, East Siberian Institute of Medical and Ecological Research, Angarsk, 665827, Russian Federation

e-mail: alexeenko85@mail.ru

alexeenko85@mail.ruhttps://orcid.org/0000-0002-9961-6408ЖурбаОльга МихайловнаZhurbaOlga M.

Доктор биол. наук, вед. науч. сотр., зав. лаб. аналитической экотоксикологии и биомониторинга ФГБНУ ВСИМЭИ, 665827, Ангарск, Россия

e-mail: zhurba99@gmail.com

DSc (Biology), leading researcher, head, Laboratory of analytical ecotoxicology and biomonitoring, East Siberian Institute of Medical and Ecological Research, Angarsk, 665827, Russian Federation

e-mail: zhurba99@gmail.com 

zhurba99@gmail.comhttps://orcid.org/0000-0001-7848-6432МериновАлексей ВладимировичMerinovAleksey V.

Канд. биол. наук, науч. сотр. ФГБНУ ВСИМЭИ, 665827, Ангарск, Россия

e-mail: alek-merinov@mail.ru

PhD (Biology), researcher, East Siberian Institute of Medical and Ecological Research, Angarsk, 665827, Russian Federation

e-mail: alek-merinov@mail.ru

alek-merinov@mail.ruhttps://orcid.org/0000-0001-8740-3133ШаяхметовСалим ФайзыевичShayahmetovSalim F.

Доктор мед. наук, профессор, вед. науч. сотр. ФГБНУ ВСИМЭИ, 665827, Ангарск, Россия

e-mail: salimf53@mail.ru

DSc (Medicine), professor, leading researcher, East Siberian Institute of Medical and Ecological Research, Angarsk, 665827, Russian Federation

e-mail: salimf53@mail.ru

salimf53@mail.ruФГБНУ «Восточно-Сибирский институт медико-экологических исследований»РоссияEast-Siberian Institute of Medical and Ecological ResearchRussian Federation202522102025104912091215Copyright © Алексеенко А.Н., Журба О.М., Меринов А.В., Шаяхметов С.Ф., 20252025Алексеенко А.Н., Журба О.М., Меринов А.В., Шаяхметов С.Ф.Alekseenko A.N., Zhurba O.M., Merinov A.V., Shayahmetov S.F.Данная работа распространяется под лицензией Creative Commons Attribution 4.0.This work is licensed under a Creative Commons Attribution 4.0 License.https://www.rjhas.ru/jour/article/view/5164Введение

Введение. Полиароматические углеводороды (ПАУ) метаболизируются в гидроксильные производные, которые выводятся из организма человека с мочой. Газовая хроматография с масс-селективным детектированием – наиболее чувствительный и эффективный метод для разделения и последующего определения содержания гидроксильных ПАУ.

Материалы и методы

Материалы и методы. В исследовании использовали газовый хроматограф с масс-селективным детектором и капиллярной колонкой HP-5MS. Извлечение проводили с помощью дисперсионной жидкостно-жидкостной микроэкстракции, экстрагирующим растворителем служил хлороформ, а диспергирующим растворителем – этанол. Дериватизацию гидроксильных ПАУ выполняли в онлайн-режиме, одновременно вводя в испаритель хроматографа хлороформный экстракт и силилирующий реагент.

Результаты

Результаты. Рассчитаны характеристики разделения гидроксильных ПАУ в виде триметилсиланов на капиллярной колонке: эффективность, коэффициент разделения, разрешение. Исследовано влияние температуры испарительного порта на сигнал детектора. Осуществлён подбор диспергирующего растворителя (этанола, ацетона, ацетонитрила, метанола) для изучения его влияния на степень экстракции. Максимальная степень экстракции отмечена при применении этанола. Исследованы закономерности изменения степени экстракции от объёма этанола и времени встряхивания. Оценены следующие метрологические характеристики: повторяемость, воспроизводимость, полнота извлечения, точность.

Ограничения исследования

Ограничения исследования. Совершенствование методического похода к измерению гидроксильных ПАУ в моче проведено без апробации метода на реальных биопробах работников, подвергающихся воздействию токсикантов в производственных условиях.

Заключение

Заключение. В усовершенствованном методическом подходе к определению содержания гидроксильных ПАУ методом газовой хроматографии с масс-селективным детектированием использование дисперсионной жидкостно-жидкостной микроэкстракции вместо классической жидкостной экстракции ускорило пробоподготовку, исключило стадию упаривания экстракта. Степень экстракции составила 70–100%. Дериватизация реагентом силил-991 в испарительном порте обеспечила экспрессность реакции.

Соблюдение этических стандартов. Исследование не требует представления комитета по биомедицинской этике или иных документов.

Участие авторов

Участие авторов: Алексеенко А.Н. – концепция и дизайн исследования, поиск источников литературы, обработка данных, написание текста; Журба О.М – сбор и обработка материала, лабораторные исследования; Меринов А.В. – сбор данных литературы, статистическая обработка, написание текста; Шаяхметов С.Ф. – организация исследования, редактирование, обсуждение результатов. Все соавторы – утверждение окончательного варианта статьи, ответственность за целостность всех её частей.

Конфликт интересов

Конфликт интересов. Авторы декларируют отсутствие явных и потенциальных конфликтов интересов в связи с публикацией данной статьи.

Финансирование

Финансирование. Работа выполнена в рамках средств, выделяемых для выполнения государственного задания ФГБНУ ВСИМЭИ.

Поступила

Поступила: 25.04.2025 / Поступила после доработки: 13.08.2025 / Принята к печати: 19.09.2025 / Опубликована: 20.10.2025

Introduction

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

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

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

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

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

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

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

Funding

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

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

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The authors declare that there are no conflicts of interest present.