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van Bekkum, Y.M. et al.

Determination of hemoglobin adducts following oral administration of 1-nitropyrene to rats using gas chromatography-tandem mass spectrometry.

1-Nitropyrene (1-NP) has successfully been used as a marker for environmental monitoring of exposure to diesel exhaust. This study presents a sensitive and selective method for detection of Hb adducts after oral administration of a single dose 1-NP to rats, by measuring 1-aminopyrene (1-AP) after in vitro hydrolysis of the adducts. Released 1-AP was extracted with hexane and derivatized with heptafluorobutyric acid anhydride prior to GC-MS-MS analysis. Optimal conditions for the release of 1-AP were hydrolysis under nitrogen, in 1 M NaOH at 70 degrees C for 60 min. Analysis of a stock solution of Hb adducts of 1-NP utilizing these conditions showed to be reproducible over a period of several weeks with a coefficient of variance of 9.5%. The determination limit was 10-20 pg 1-AP per 70-90 mg globin. A study of the time course of Hb adduct formation showed a fast absorption and an early peak concentration of released 1-AP, approximately 39 pg 1-AP/mg globin at 3 h after exposure. After the maximum was reached, 1-AP concentrations decreased bi-phasically. Initially a fast decline was observed, followed by a slow decrease to 5.9+/-1.9 pg 1-AP/mg globin at 24 h after administration.[1]

References

Determination of hemoglobin adducts following oral administration of 1-nitropyrene to rats using gas chromatography-tandem mass spectrometry. van Bekkum, Y.M., Scheepers, P.T., van den Broek, P.H., Velders, D.D., Noordhoek, J., Bos, R.P. J. Chromatogr. B Biomed. Sci. Appl. (1997) [Pubmed]

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