Regular Article

Journal of Investigative Dermatology (2001) 117, 263–268; doi:10.1046/j.1523-1747.2001.01416.x

Cyclobutane Thymidine Dimers are Present in Human Urine Following Sun Exposure: Quantitation Using 32P-Postlabeling and High-Performance Liquid Chromatography

Frank Le Curieux and Kari Hemminki

Department of Biosciences at Novum, Karolinska Institute, Huddinge, Sweden

Correspondence: Dr Kari Hemminki, Department of Biosciences at Novum, Karolinska Institute, 141 57 Huddinge, Sweden. Email: kari.hemminki@cnt.ki.se

Received 27 June 2000; Revised 30 March 2001; Accepted 2 April 2001.

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Abstract

Cyclobutane thymidine dimer (T=T) is the major DNA photoproduct formed in human skin after solar radiation. We have developed a 32P-postlabeling method suitable for quantitating T=T in human urine with a detection limit of about 0.5 fmol per 10 mul urine. The method was used in the present study to measure the daily T=T urinary level of two volunteers over a 15 d period, including frequent sun exposures ranging from 0 to 5 h daily. T=T was not detected before or immediately (4 h) after the initial sun exposure but was first observed in urine samples collected 18 h after the initial exposure. Thereafter, urinary T=T levels gradually increased up to a peak reached about 3 d after the maximum sun exposure. The levels decreased during the following days but were still detectable 8 d after the last sun exposure. About 70–75% decrease in excreted T=T was observed after 8 d. The T=T levels measured in urine were lower but in the same order of magnitude as the levels expected after a theoretical calculation based on previous published results and reasonable assumptions. This study shows the occurrence of cyclobutane thymidine dimers in human urine after skin exposure to solar radiation.

Keywords:

32P-postlabeling, creatinine, cyclobutane thymidine dimers, high-performance liquid chromatography, human urine, photoproduct, sun exposure, ultraviolet radiation

Abbreviations:

T=T, cyclobutane thymidine dimer; TpT, parent thymidine dimer

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