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摘要
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 C-18 column (150 mm x 4.6 mm, 5 mu m) 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 lambda (ex)/lambda (em) = 365/450 nm and lambda (ex)/lambda (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 +/- A 5.0-113.5 +/- A 6.7 % for 3-OHB[a]P and 79.3 +/- A 6.2-141.2 +/- A 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.
- 出版日期2015-5
- 单位大连医科大学
