New Rapid HPLC Method for Separation and Determination of Benzo[A]Pyrene Hydroxyderivatives

Abstract

A new rapid, one-step semi-quantitative HPLC-MS method applicable for the separation and characterization of the group of total 10 BaP hydroxyderivatives (1-hydroxy benzo[a]pyrene, 2-hydroxybenzo[a]pyrene, 3-hydroxybenzo[a]pyrene, 4-hydroxybenzo [a]pyrene, 7-hydroxybenzo[a]pyrene, 8-hydroxybenzo[a]pyrene, 9-hydroxybenzo[a]py rene, 10-hydroxybenzo[a]pyrene, 12-hydroxybenzo[a]pyrene and benzo[a]pyrene-9,10-dihydrodiol) was developed, involving electrospray ionization technique (ESI) in positive mode. Its qualitative and quantitative characteristics were selected by the analysis of the model OH-BaP mixture consisted of compounds mentioned above. The model system offered the linear response in the whole concentration range used (50–1000 ng.mL⁻¹) as well as the resolution sufficient for semi-quantitative purposes (R_S > 0.5). Subsequently, the suitability of the developed method was tested on BaP decomposition products to be formed during photooxidation at λ = 365 nm. These analyses confirmed the applicability of the proposed method, succeeding in the identification of 5 BaP hydroxyderivatives formed upon the BaP photooxidation experiment.

Key Words:

• benzo[a]pyrene
• ESI
• HPLC-MS
• hydroxyderivatives
• photooxidation

Acknowledgments

This article is the result of the projects “Establishment of a HiTech Centre for Research on Formation, Elimination and Assessment of Contaminants in Food” and “Centre of Excellence for Contaminants and Microorganisms in Food” implementation, both supported by the Research & Development Operational Programme funded by the ERDF. This work was also supported by the Slovak Research and Development Agency under the contract no. APVV-0602-07.