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Determination of 3-OHB[a]P and (+)-anti-BPDE in Rats Blood and Brain Tissue of B[a]P Exposure by HPLC with Fluorescence Detection

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Abstract

The aim of this paper was to develop an analytical method for determining the levels of the Benzo[a]pyrene (B[a]P) metabolites, 3-hydroxybenzo(a)pyrene (3-OHB[a]P), and (+)-anti-benzo(a)pyrene diol-epoxide( BPDE, combined with DNA to formate adducts) in the blood and brain tissue of B[a]P-exposed rats using high-performance liquid chromatography coupled with fluorescence detection (HPLC/FD). The chromatographic separation was performed with a CenturySIL BDS C18 column (150 mm × 4.6 mm, 5 μm) using mobile phases consisted of different methanol–water ratios [97:3 or 55:45 (v/v)] and different pHs (4.5 or 7.0). The amounts of 3-OHB[a]P and B[a]P-tetrols I-1 released after acid hydrolysis of (+)-anti-BPDE were detected by fluorescence at λ ex/λ em = 365/450 nm and λ ex/λ em = 245/395 nm. The calibration curves were linear (r 2 > 0.9990) for 3-OHB[a]P within 27–4320 ng mL−1 and for (+)-anti-BPDE within 1–160 ng mL−1, and the limits of detection (S/N = 3) of 3-OHB[a]P and (+)-anti-BPDE were 0.2 and 0.3 ng mL−1, respectively. Variation in intra- and inter-day assay as well as the data concerned with the stability of these compounds indicated no significant degradation under current experimental conditions. The average recoveries of the compound were in the range of 73.6 ± 5.0–113.5 ± 6.7 % for 3-OHB[a]P and 79.3 ± 6.2–141.2 ± 13.8 % for (+)-anti-BPDE. Our study have provided a sensitive, reliable, and rapid HPLC/FD-based method for the analysis of 3-OHB[a]P and (+)-anti-BPDE in rats blood and brain tissue.

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Acknowledgments

This work is financially supported by the National Natural Science Foundation of China (30770724), the National Quality Inspection Science Foundation of China (L2013344), and Liaoning Provincial Department of Education Research Foundation of China (2013IK228).

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    Authors and Affiliations

    1. College of Medical Laboratory, Dalian Medical University, Liaoning, Dalian, 116044, LiaoNing Province, China

      Xiao-Chen Liu, Lin Qi, Nan Ma, Guang Yang, Na Liu, Hui Liu & Fa-Sheng Li

    2. Department of Clinical Laboratory, Second Affiliated Hospital of Dalian Medical University, Dalian, 116044, LiaoNing Province, China

      Xiao-Chen Liu

    3. College of Pharmacy, Washington State University, Spokane, WA, 99210-1495, USA

      Gang Chen

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    Correspondence to Fa-Sheng Li.

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    X.-C. Liu, L. Qi, N. Ma contributed equally to this work.

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    Liu, XC., Qi, L., Ma, N. et al. Determination of 3-OHB[a]P and (+)-anti-BPDE in Rats Blood and Brain Tissue of B[a]P Exposure by HPLC with Fluorescence Detection. Chromatographia 78, 663–673 (2015). https://doi.org/10.1007/s10337-015-2890-2

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