medlitГигиена и санитарияHygiene and Sanitation0016-99002412-0650Federal Scientific Center of Hygiene named after F.F. Erisman10.47470/0016-9900-2020-99-10-1153-1158medlit-1080Research ArticleМЕТОДЫ ГИГИЕНИЧЕСКИХ ИССЛЕДОВАНИЙMETHODS OF HYGIENIC AND EXPERIMENTAL INVESTIGATIONSОптимизация условий пробоподготовки с помощью математического планирования для определения 1-гидроксипирена в моче методом газовой хромато-масс-спектрометрииOptimization of conditions for the sample preparation using mathematical planning for determination of 1-hydroxypyrene in urine using method of gas chromatography-mass spectrometryhttps://orcid.org/0000-0003-4980-5304АлексеенкоАнтон НиколаевичAlekseenkoAnton N.

Канд. хим. наук, ст. науч. сотр. лаб. аналитической экотоксикологии и биомониторинга, ФГБНУ «Восточно-Сибирский институт медико-экологических исследований», 665827, Ангарск.

e-mail: alexeenko85@mail.ru

MD, Ph.D., senior research, Laboratory of analytical ecotoxicology and biomonitoring of the East-Siberian Institution of Medical and Ecological Research, Angarsk, 665827, Russian Federation.

e-mail: labchem99@gmail.com

alexeenko85@mail.ruhttps://orcid.org/0000-0002-9961-6408ЖурбаО. М.ZhurbaOlga M.noemail@neicon.ruhttps://orcid.org/0000-0001-7848-6432МериновА. В.MerinovAlexey V.noemail@neicon.ruhttps://orcid.org/0000-0001-8740-3133ШаяхметовС. Ф.ShayakhmetovSalim F.noemail@neicon.ruФГБНУ «Восточно-Сибирский институт медико-экологических исследований»East-Siberian Institute of Medical and Ecological Research202001122020991011531158Copyright © Алексеенко А.Н., Журба О.М., Меринов А.В., Шаяхметов С.Ф., 20202020Алексеенко А.Н., Журба О.М., Меринов А.В., Шаяхметов С.Ф.Alekseenko A.N., Zhurba O.M., Merinov A.V., Shayakhmetov 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/1080Введение

Введение. Для оценки воздействия ПАУ на человека наибольшее распространение получил биологический мониторинг 1-гидроксипирена. Определение 1-гидроксипирена в моче осуществляется с помощью газовой хромато-масс-спектрометрии (ГХ-МС).

Материал и методы

Материал и методы. Пробоподготовка заключается в извлечении аналита из биологической матрицы методом 2-кратной жидкостной экстракции гексаном, упаривании экстракта до сухого остатка в токе азота, перерастворении сухого остатка в силилирующем агенте БСТФА. В работе использовали газовый хроматограф Agilent 7890A с капиллярной колонкой HP-5MS и масс-селективным детектором. Идентифицировали 1-гидроксипирен в виде производного (триметилсилана) на масс-хроматограмме по времени удерживания и соотношению интенсивностей регистрируемых ионов.

Результаты

Результаты. Оптимизацию условий жидкостной экстракции 1-гидроксипирена осуществляли с помощью математического планирования, варьируя массу сульфата магния, время экстракции, кратность экстракции. Матрица планирования включала в себя восемь опытов, в качестве параметра оптимизации служила степень извлечения 1-гидроксипирена. Интерпретация модели показала, что кратность экстракции вносит больший вклад в формирование степени извлечения, чем масса сульфата магния и время экстракции.

Заключение

Заключение. Выбор оптимальных условий жидкостной экстракции 1-гидроксипирена методом математического планирования позволил оптимизировать методику определения данного аналита в моче методом ГХ-МС для проведения биологического мониторинга.

Introduction

Introduction. Biological monitoring of 1-hydroxypyrene is the most widely used method for evaluating the effects of PAHs on humans. The determination of 1-hydroxypyrene in urine is performed using gas chromatography-mass spectrometry (GC-MS).

Material and methods

Material and methods. Sample preparation consists of extracting the analyte from the biological matrix by 2-fold liquid extraction with hexane, evaporation the extract to the dry residue in the nitrogen current, and re-dissolution of the dry residue in the silylating agent BSTFA. We used gas chromatograph Agilent 7890A with an HP-5MS capillary column and a mass-selective detector. 1-hydroxypyrene was identified as trimethyl silane on a mass chromatogram based on the retention time and intensity ratio of the registered ions.

Results

Results. Optimization of the conditions for liquid extraction of 1-hydroxypyrene was performed using mathematical planning, varying the mass of magnesium sulfate, the extraction time, and the extraction multiplicity. The planning matrix included eight experiments, and the degree extraction of 1-hydroxypyrene was used as an optimization parameter. Interpretation of the model showed the multiplicity of extraction to contribute more to the formation of the degree of extraction than the mass of magnesium sulfate and the extraction time.

Discussion

Discussion. The proposed method foк the sample preparation, based on the extraction of 1-hydroxypyrene by 2-fold liquid extraction with hexane for 2 min with the addition of 0.5 g of magnesium sulfate to the biological sample, allowed reaching the detection limit of 0.1 ng/ml. The evaluation of metrological characteristics showed RSD of the reproducibility to do not exceed 6.4%, the systematic error is not significant, and the accuracy index in the form of a total error is not higher than 15%. Levels of 1-hydroxypyrene in the urine of aluminum production workers are 53 - 414 times higher than in the control group.

Conclusion

Conclusion. The choice of optimal conditions for liquid extraction of 1-hydroxypyrene by mathematical planning allowed developing a method for the determination this analyte in urine by GC-MS for biological monitoring.

1-гидроксипирен в мочегазовая хромато-масс-спектрометрияматематическое планированиеоптимальные условияжидкостная экстракция1-hydroxypyrene in urinegas chromatography-mass spectrometrymathematical planningoptimal conditionsliquid extractionReferencesBrandt H.C., Watson W.P. Monitoring human occupational and environmental exposures to polycyclic aromatic compounds. Ann. Occup. Hyg. 2003; 47(5): 349-78. https://doi.org/10.1093/annhyg/meg052Brandt H.C., Watson W.P. Monitoring human occupational and environmental exposures to polycyclic aromatic compounds. Ann. Occup. Hyg. 2003; 47(5): 349–78. https://doi.org/10.1093/annhyg/meg052Jacob J., Seidel A. Biomonitoring of polycyclic aromatic hydrocarbons in human urine. J. Chromatogr. B Analyt. Technol. Biomed. Life Sci. 2002; 778(1-2): 31-47. https://doi.org/10.1016/s0378-4347(01)00467-4Jacob J., Seidel A. Biomonitoring of polycyclic aromatic hydrocarbons in human urine. J. Chromatogr. B Analyt. Technol. Biomed. Life Sci. 2002; 778(1–2): 31–47. https://doi.org/10.1016/s0378-4347(01)00467-4Bosetti C., Boffetta, P., La Vecchia C. Occupational exposures to polycyclic aromatic hydrocarbons, and respiratory and urinary tract cancers: a quantitative review to 2005. Ann. Oncol. 2005; 18(3): 431-46. https://doi.org/10.1093/annonc/mdl172Bosetti C., Boffetta, P., La Vecchia C. Occupational exposures to polycyclic aromatic hydrocarbons, and respiratory and urinary tract cancers: a quantitative review to 2005. Ann. Oncol. 2005; 18(3): 431–46. https://doi.org/10.1093/annonc/mdl172Al-Saleh I., Alsabbahen A., Shinwari N., Billedo G., Mashhour A., Al-Sarraj Y., et al. Polycyclic aromatic hydrocarbons (PAHs) as determinants of various anthropometric measures of birth outcome. Sci. Total Environ. 2013; 444: 565-78. https://doi.org/10.1016/j.scitotenv.2012.12.021Al-Saleh I., Alsabbahen A., Shinwari N., Billedo G., Mashhour A., Al-Sarraj Y., et al. Polycyclic aromatic hydrocarbons (PAHs) as determinants of various anthropometric measures of birth outcome. Sci. Total Environ. 2013; 444: 565–78. https://doi.org/10.1016/j.scitotenv.2012.12.021Barbeau D., Persoons R., Marques M., Hervé C., Laffitte-Rigaud G., Maitre A. Relevance of urinary 3-hydroxybenzo(a)pyrene and 1-hydroxypyrene to assess exposure to carcinogenic polycyclic aromatic hydrocarbon mixtures in metallurgy workers. Ann. Occup. Hyg. 2014; 58(5): 579-90. https://doi.org/10.1093/annhyg/meu004Barbeau D., Persoons R., Marques M., Hervé C., Laffitte-Rigaud G., Maitre A. Relevance of urinary 3-hydroxybenzo(a)pyrene and 1-hydroxypyrene to assess exposure to carcinogenic polycyclic aromatic hydrocarbon mixtures in metallurgy workers. Ann. Occup. Hyg. 2014; 58(5): 579–90. https://doi.org/10.1093/annhyg/meu004Klöslová Z., Drímal M., Balog K., Koppová K., Dubajová J. The relations between polycyclic aromatic hydrocarbons exposure and 1-OHP levels as a biomarker of the exposure. Cent. Eur. J. Public Health. 2016; 24(4): 302-7. https://doi.org/10.21101/cejph.a4179Klöslová Z., Drímal M., Balog K., Koppová K., Dubajová J. The relations between polycyclic aromatic hydrocarbons exposure and 1-OHP levels as a biomarker of the exposure. Cent. Eur. J. Public Health. 2016; 24(4): 302–7. https://doi.org/10.21101/cejph.a4179Зибарев Е.В., Эллингсен Д.Г., Томассен И., Чащин В.П., Чащин М.В., Кузьмин А.В. Биологический мониторинг как способ управления профессиональными рисками. Уральский медицинский журнал. 2011; (9): 16-8Zibarev E.V., Ellingsen D.G., Tomassen I., Chashchin V.P., Chashchin M.V., Kuz’min A.V. Biological monitoring as method occupational risk assessment. Ural’skiy meditsinskiy zhurnal. 2011; (9): 16–8. (in Russian)Grover P.L. Pathways involved in the metabolism and activation of polycyclic hydrocarbons. Xenobiotica. 1986; 16(10-11): 915-31. https://doi.org/10.3109/00498258609038974Grover P.L. Pathways involved in the metabolism and activation of polycyclic hydrocarbons. Xenobiotica. 1986; 16(10–11): 915–31. https://doi.org/10.3109/00498258609038974Unwin J., Cocker J., Scobbie E., Chambers H. An assesment of occupational exposure to polycyclic aromatic hydrocarbonsin the UK. Ann. Occup. Hyg. 2006; 50(4): 395-403. https://doi.org/10.1093/annhyg/mel010Unwin J., Cocker J., Scobbie E., Chambers H. An assesment of occupational exposure to polycyclic aromatic hydrocarbonsin the UK. Ann. Occup. Hyg. 2006; 50(4): 395–403. https://doi.org/10.1093/annhyg/mel010Boogaard P.J. Urinary biomarkers in the risk assessment of PAHs. Occup. Environ. Med. 2008; 65(4): 221-2. https://doi.org/10.1136/oem.2007.034157Boogaard P.J. Urinary biomarkers in the risk assessment of PAHs. Occup. Environ. Med. 2008; 65(4): 221–2. https://doi.org/10.1136/oem.2007.034157Hansen A., Mathiesen L., Pedersen M., Knudsen L.E. Urinary 1-hydroxypyrene (1-HP) in environmental and occupational studies - a review. Int. J. Hyg. Environ. Health. 2008; 211(5-6): 471-503. https://doi.org/10.1016/j.ijheh.2007.09.012Hansen A., Mathiesen L., Pedersen M., Knudsen L.E. Urinary 1-hydroxypyrene (1-HP) in environmental and occupational studies – a review. Int. J. Hyg. Environ. Health. 2008; 211(5–6): 471–503. https://doi.org/10.1016/j.ijheh.2007.09.012Rossella F., Campo L., Pavanello S., Kapka L., Siwinska E., Fustinoni S. Urinary polycyclic aromatic hydrocarbons and monohydroxy metabolites as biomarkers of exposure in coke oven workers. Occup. Environ. Med. 2009; 66(8): 509-16. https://doi.org/10.1136/oem.2008.042796Rossella F., Campo L., Pavanello S., Kapka L., Siwinska E., Fustinoni S. Urinary polycyclic aromatic hydrocarbons and monohydroxy metabolites as biomarkers of exposure in coke oven workers. Occup. Environ. Med. 2009; 66(8): 509–16. https://doi.org/10.1136/oem.2008.042796Leroyer A., Jeandel F., Maitre A., Howsam M., Deplanque D., Mazzuca M., et al. 1-Hydroxypyrene and 3-hydroxybenzo[a]pyrene as biomarkers of exposure to PAH in various environmental exposure situations. Sci. Total Environ. 2010; 408(5): 1166-73. https://doi.org/10.1016/j.scitotenv.2009.10.073Leroyer A., Jeandel F., Maitre A., Howsam M., Deplanque D., Mazzuca M., et al. 1-Hydroxypyrene and 3-hydroxybenzo[a]pyrene as biomarkers of exposure to PAH in various environmental exposure situations. Sci. Total Environ. 2010; 408(5): 1166–73. https://doi.org/10.1016/j.scitotenv.2009.10.073Hadrup N., Mielżyńska-Švach D., Kozłowska A., Campisi M., Pavanello S., Vogel U. Association between a urinary biomarker for exposure to PAH and blood level of the acute phase protein serum amyloid A in coke oven workers. Environ. Health. 2019; 18(1): 81. https://doi.org/10.1186/s12940-019-0523-1Hadrup N., Mielżyńska-Švach D., Kozłowska A., Campisi M., Pavanello S., Vogel U. Association between a urinary biomarker for exposure to PAH and blood level of the acute phase protein serum amyloid A in coke oven workers. Environ. Health. 2019; 18(1): 81. https://doi.org/10.1186/s12940-019-0523-1Samir A.M., Shaker D.A., Fathy M.M., Hafez S.F., Abdullatif M.M., Rashed L.A., et al. Urinary and genetic biomonitoring of polycyclic aromatic hydrocarbons in egyptian coke oven workers: associations between exposure, effect, and carcinogenic risk assessment. Int. J. Occup. Environ. Med. 2019; 10(3): 124-36. https://doi.org/10.15171/ijoem.2019.1541Samir A.M., Shaker D.A., Fathy M.M., Hafez S.F., Abdullatif M.M., Rashed L.A., et al. Urinary and genetic biomonitoring of polycyclic aromatic hydrocarbons in egyptian coke oven workers: associations between exposure, effect, and carcinogenic risk assessment. Int. J. Occup. Environ. Med. 2019; 10(3): 124–36. https://doi.org/10.15171/ijoem.2019.1541Du M., Mullins B.J., Franklin P., Musk A.W., Elliot N.S.J., Sodhi-Berry N., et al. Measurement of urinary 1-aminopyrene and 1-hydroxypyrene as biomarkers of exposure to diesel particulate matter in gold miners. Sci. Total Environ. 2019; 685: 723-8. https://doi.org/10.1016/j.scitotenv.2019.06.242Du M., Mullins B.J., Franklin P., Musk A.W., Elliot N.S.J., Sodhi-Berry N., et al. Measurement of urinary 1-aminopyrene and 1-hydroxypyrene as biomarkers of exposure to diesel particulate matter in gold miners. Sci. Total Environ. 2019; 685: 723–8. https://doi.org/10.1016/j.scitotenv.2019.06.242Adetona A.M., Martin W.K., Warren S.H., Hanley N.M., Adetona O., Zhang J.J., et al. Urinary mutagenicity and other biomarkers of occupational smoke exposure of wildland firefighters and oxidative stress. Inhal. Toxicol. 2019; 31(2): 73-87. https://doi.org/10.1080/08958378.2019.1600079Adetona A.M., Martin W.K., Warren S.H., Hanley N.M., Adetona O., Zhang J.J., et al. Urinary mutagenicity and other biomarkers of occupational smoke exposure of wildland firefighters and oxidative stress. Inhal. Toxicol. 2019; 31(2): 73–87. https://doi.org/10.1080/08958378.2019.1600079Vimercati L., Bisceglia L., Cavone D., Caputi A., De Maria L, Delfino M., et.al. Environmental monitoring of PAHs exposure, biomarkers and vital status in coke oven workers. Int. J. Environ. Res. Public Health. 2020; 17(7): 2199. https://doi.org/10.3390/ijerph17072199Vimercati L., Bisceglia L., Cavone D., Caputi A., De Maria L, Delfino M., et.al. Environmental monitoring of PAHs exposure, biomarkers and vital status in coke oven workers. Int. J. Environ. Res. Public Health. 2020; 17(7): 2199. https://doi.org/10.3390/ijerph17072199Ratelle M., Khoury C., Adlard B., Laird B. Polycyclic aromatic hydrocarbons (PAHs) levels in urine samples collected in a subarctic region of the Northwest Territories, Canada. Environ. Res. 2020; 182: 109112. https://doi.org/10.1016/j.envres.2020.109112Ratelle M., Khoury C., Adlard B., Laird B. Polycyclic aromatic hydrocarbons (PAHs) levels in urine samples collected in a subarctic region of the Northwest Territories, Canada. Environ. Res. 2020; 182: 109112. https://doi.org/10.1016/j.envres.2020.109112Cao L., Wang D., Wen Y., He H., Chen A., Hu D., et al. Effects of environmental and lifestyle exposures on urinary levels of polycyclic aromatic hydrocarbon metabolites: A cross-sectional study of urban adults in China. Chemosphere. 2020; 240: 124898. https://doi.org/10.1016/j.chemosphere.2019.124898Cao L., Wang D., Wen Y., He H., Chen A., Hu D., et al. Effects of environmental and lifestyle exposures on urinary levels of polycyclic aromatic hydrocarbon metabolites: A cross-sectional study of urban adults in China. Chemosphere. 2020; 240: 124898. https://doi.org/10.1016/j.chemosphere.2019.124898Oliveira M., Costa S., Vaz J., Fernandes A., Slezakova K., Delerue-Matos C., et al. Firefighters exposure to fire emissions: Impact on levels of biomarkers of exposure to polycyclic aromatic hydrocarbons and genotoxic/oxidative-effects. J. Hazard. Mater. 2020; 383: 121179. https://doi.org/10.1016/j.jhazmat.2019.121179Oliveira M., Costa S., Vaz J., Fernandes A., Slezakova K., Delerue-Matos C., et al. Firefighters exposure to fire emissions: Impact on levels of biomarkers of exposure to polycyclic aromatic hydrocarbons and genotoxic/oxidative-effects. J. Hazard. Mater. 2020; 383: 121179. https://doi.org/10.1016/j.jhazmat.2019.121179Blau K., Halket J.M. Handbook of Derivates for Chromatography. New-York: Wiley; 1993.Blau K., Halket J.M. Handbook of Derivates for Chromatography. New-York: Wiley; 1993.Schummer C., Delhomme O., Appenzeller B.M., Wennig R., Millet M. Comparison of MTBSTFA and BSTFA in derivatization reaction of polar compounds prior to GC/MS analysis. Talanta. 2009; 77(4): 1473-82. https://doi.org/10.1016/j.talanta.2008.09.043Schummer C., Delhomme O., Appenzeller B.M., Wennig R., Millet M. Comparison of MTBSTFA and BSTFA in derivatization reaction of polar compounds prior to GC/MS analysis. Talanta. 2009; 77(4): 1473–82. https://doi.org/10.1016/j.talanta.2008.09.043Shin H.S., Lim H.H. Simultaneous determination of 2-naphtol and 1-hydroxy pyrene in urine by gas chromatography-mass spectrometry. J. Chromatogr. B Analyt. Technol. Biomed. Life Sci. 2011; 879(7-8): 489-94. https://doi.org/10.1016/j.jchromb.2011.01.009Shin H.S., Lim H.H. Simultaneous determination of 2-naphtol and 1-hydroxy pyrene in urine by gas chromatography-mass spectrometry. J. Chromatogr. B Analyt. Technol. Biomed. Life Sci. 2011; 879(7–8): 489–94. https://doi.org/10.1016/j.jchromb.2011.01.009Chen D., Xu H. Simultaneous HPLC-MS determination of 8-hydroxy-2’-deoxyguanosine, 3-hydroxyphenanthrene and 1-hydroxypyrene after online in-tube solid phase microextraction using a graphene oxide/poly(3,4-ethylenedioxythiophene)/polypyrrole composite. Mikrochim. Acta. 2019; 186(5): 300. https://doi.org/10.1007/s00604-019-3429-2Chen D., Xu H. Simultaneous HPLC-MS determination of 8-hydroxy-2’-deoxyguanosine, 3-hydroxyphenanthrene and 1-hydroxypyrene after online in-tube solid phase microextraction using a graphene oxide/poly(3,4-ethylenedioxythiophene)/polypyrrole composite. Mikrochim. Acta. 2019; 186(5): 300. https://doi.org/10.1007/s00604-019-3429-2Martin-Tornero E., Luque-Uría A., Durán-Merás I., Espinosa-Mansilla A. A novel analytical methodology for the determination of hydroxy polycyclic aromatic hydrocarbons in breast and cow milk samples. J. Chromatogr. B Analyt. Technol. Biomed. Life Sci. 2020; 1136: 121912. https://doi.org/10.1016/j.jchromb.2019.121912Martin-Tornero E., Luque-Uría A., Durán-Merás I., Espinosa-Mansilla A. A novel analytical methodology for the determination of hydroxy polycyclic aromatic hydrocarbons in breast and cow milk samples. J. Chromatogr. B Analyt. Technol. Biomed. Life Sci. 2020; 1136: 121912. https://doi.org/10.1016/j.jchromb.2019.121912Вершинин В.И., Перцев Н.В. Планирование и математическая обработка результатов химического эксперимента. Омск; 2005.Vershinin V.I., Pertsev N.V. Planning and Mathematical Processing of Chemical Experiment Results [Planirovanie i matematicheskaya obrabotka rezul’tatov khimicheskogo eksperimenta]. Omsk; 2005. (in Russian)Адлер Ю.П., Маркова Е.В., Грановский Ю.А. Планирование эксперимента при поиске оптимальных условий. М.: Наука; 1971.Adler Yu.P., Markova E.V., Granovskiy Yu.A. Planning an Experiment when Searching for Optimal Conditions [Planirovanie eksperimenta pri poiske optimal’nykh usloviy]. Moscow: Nauka; 1971. (in Russian)Смагунова А.Н., Пашкова Г.В., Белых Л.И. Математическое планирование эксперимента в методических исследованиях аналитической химии. Иркутск; 2015.Smagunova A.N., Pashkova G.V., Belykh L.I. Mathematical Planning of an Experiment in Methodological Research of Analytical Chemistry [Matematicheskoe planirovanie eksperimenta v metodicheskikh issledovaniyakh analiticheskoy khimii]. Irkutsk; 2015. (in Russian)Смагунова А.Н., Карпукова О.М. Методы математической статистики в аналитической химии. Ростов-на-Дону: Феникс; 2012.Smagunova A.N., Karpukova O.M. Methods of Mathematical Statistics in Analytical Chemistry [Metody matematicheskoy statistiki v analiticheskoy khimii]. Rostov-na-Donu: Feniks; 2012. (in Russian)

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