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Benzo[a]pyrene-specific online high-performance liquid chromatography fractionation of air particulate extracts–A tool for evaluating biological interactions
Stockholm University, Faculty of Science, Department of Analytical Chemistry.
Stockholm University, Faculty of Science, Department of Analytical Chemistry.
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2014 (English)In: Journal of Chromatography A, ISSN 0021-9673, E-ISSN 1873-3778, Vol. 1355, p. 100-106Article in journal (Refereed) Published
Abstract [en]

Benzo[a]pyrene (B[a]P) is a known human carcinogen and is commonly used as a surrogate for assessing the carcinogenic risk posed by complex mixtures of polycyclic aromatic hydrocarbons (PAHs) present in air particulate matter (PM). However, studies have shown that using B[a]P as a surrogate may underestimate the carcinogenic potential of PAH mixtures, as the risk assessment approach does not consider interaction effects. Thus, toxicological studies using B[a]P to assess its carcinogenic potential in environmentally derived complex mixtures, as opposed to single compound experiments, could improve risk assessment. The intention of the present study was to develop an online HPLC fractionation system for the selective removal of B[a]P from air PM extracts. Two serial pyrenylethyl (PYE) columns enabled selective separation of B[a]P from its isomers and other PAHs as well as a short fractionation cycle of 30 minutes. One run consisted of three collection steps: the first fraction contained PAHs eluting earlier than B[a]P, the second contained B[a]P and the last contained later-eluting PAHs. The selectivity and recovery of the system was investigated using extracts of Stockholm air PM samples. The overall recovery for all PAHs was approximately 80%, and the system proved to be selective, as it removed 94% of B[a]P and less than 3% of benzo[b]fluoranthene from the complex PAH mixture. Exposing human cells to blanks generated by the fractionation system did not induce cytotoxicity or DNA damage signalling. In conclusion, the online HPLC system was selective for B[a]P fractionation whilst minimising run-to-run variation and allowing repeated fractionations for larger samples due to its relatively short run time

Place, publisher, year, edition, pages
2014. Vol. 1355, p. 100-106
National Category
Analytical Chemistry
Research subject
Analytical Chemistry
Identifiers
URN: urn:nbn:se:su:diva-104502DOI: 10.1016/j.chroma.2014.05.082ISI: 000340302500011OAI: oai:DiVA.org:su-104502DiVA, id: diva2:723654
Available from: 2014-06-08 Created: 2014-06-11 Last updated: 2018-09-04Bibliographically approved
In thesis
1. Automation, hyphenation and multidimensional chromatography for PAH analysis: Analytical techniques to simplify sample complexity
Open this publication in new window or tab >>Automation, hyphenation and multidimensional chromatography for PAH analysis: Analytical techniques to simplify sample complexity
2018 (English)Doctoral thesis, comprehensive summary (Other academic)
Abstract [en]

One group of organic environmental pollutants which are potentially hazardous to human health is polycyclic aromatic hydrocarbons (PAHs). These compounds consist of two or more benzene rings in their structure among which benzo[a]pyrene (B[a]P) is classified as a human carcinogen. In order to generate accurate data for PAHs, reliable analytical methods are a prerequisite for PAH monitoring in the environment and cancer risk assessments. However, there are several analytical challenges in PAH analysis because they are a large number of compounds with different physical and chemical properties and occur at varying concentration levels, often in complex matrices. Thus, this thesis aimed at tackling these difficulties in PAH analysis using various analytical techniques such as automation, hyphenation and multidimensional chromatography, particularly to increase the detectability and separation efficiency of PAHs in complex matrices. Furthermore, analytical methods were also presented as a tool for toxicological studies.

A fully automated two-dimensional (2D)-liquid chromatography (LC)/2D-gas chromatography system was developed to enable the online sample clean-up, separation and detection, initially with two flame ionization detectors (FIDs) (Paper I). The introduction of mass selective detectors (MSDs) instead of two FIDs further advanced the original 2D system, improving the detectability and selectivity in PAH analysis (Paper II). The PAH levels were determined in two standard reference materials (SRMs) from the National Institute of Standards and Technology (NIST), i.e. urban dust (SRM 1649a) and diesel particulate extract (SRM 1975). The measured PAH levels were in good agreement with those obtained from FID- and MSD-based systems and reported by NIST (Paper I and II). Additionally, the PAH determination was done in particulates from wood burning, which also showed comparable results between FID and MSD systems (Paper II). The FID-based system showed slightly better separation efficiency owing to the use of hydrogen as carrier gas instead of helium and similar detectability to that of MSD (Paper I). However, the MSD-based system was advantageous to detecting late-eluting PAHs and separating interfering peaks in complex matrices (Paper II).

Paper III presented an online LC system for B[a]P-selective fractionation as a tool for studying biological interactions. Fractions could be obtained from Stockholm air particulate matter (PM) extract with or without B[a]P. The selective fractionation was achieved using serial LC columns of two pyrenyl phases, resulting in more than 90% of B[a]P in the B[a]P-containing fraction. An analytical method for identification and quantification of benz[j]aceanthrylene (B[j]A) in air PM was developed and described in paper IV. B[j]A is rarely measured due to its low abundance but is known to have high carcinogenic potential. The determination of B[j]A in air PM from Stockholm (Sweden) and Limeira (Brazil), was done together with a series of toxicological studies. The measurement and toxicological data showed an increased number of estimated cancer cases from air PM exposure when levels of B[j]A were taken into account.

In conclusion, this thesis presents various analytical approaches to obtain more accurate PAH data as well as the possibilities of using them in toxicological research.

Place, publisher, year, edition, pages
Stockholm: Department of Environmental Science and Analytical Chemistry, Stockholm University, 2018. p. 88
Keywords
polycyclic aromatic hydrocarbon, PAH, benzo[a]pyrene, B[a]P, carcinogenicity, cancer risk assessment, complex matrix, automation, hyphenation, multidimensional chromatography, two-dimensional, method translation, online fractionation, benz[j]aceanthrylene, B[j]A, relative potency factor
National Category
Analytical Chemistry
Research subject
Analytical Chemistry
Identifiers
urn:nbn:se:su:diva-159716 (URN)978-91-7797-336-2 (ISBN)978-91-7797-337-9 (ISBN)
Public defence
2018-10-19, Magnélisalen, Kemiska övningslaboratoriet, Svante Arrhenius väg 16 B, Stockholm, 10:00 (English)
Opponent
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Available from: 2018-09-26 Created: 2018-09-04 Last updated: 2018-10-22Bibliographically approved

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