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Determination of human serum 8-hydroxy-2′-deoxyguanosine (8-OHdG) by HPLC-ECD combined with solid phase extraction (SPE)
Authors:Satoshi Koide  Yuri Kinoshita  Narushi Ito  Jun Kimura  Kenji Yokoyama  Isao Karube
Institution:1. Biotechnology Industrialization Center, National Institute of Advanced Industrial Science and Technology (AIST), c/o Katayanagi Advanced Research Laboratories, Tokyo University of Technology, 1404-1 Katakura-Machi, Hachioji, Tokyo 192-0982, Japan;2. TANITA Corporation, 1-14-2 Maeno-cho, Itabashi-ku, Tokyo 174-8630, Japan;3. Tokyo University of Technology, 1404-1 Katakura-Machi, Hachioji, Tokyo 192-0982, Japan
Abstract:Human serum 8-hydroxy-2′-deoxyguanosine (8-OHdG) was measured by HPLC-ECD method combined with solid phase extraction (SPE) developed by our group: (our proprietary kit, named 8-OHdG Pre-treatment Kit (TANITA Corporation)). The major interfering substances and proteins in serum were removed by 8-OHdG Pre-treatment Kit. This measurement method was highly reproducible (CV = 2.2–7.1%) and demonstrated the lower detection limit for control serum sample of less than 10 pg/ml without the sample evaporation. The other hand 8-OHdG concentration in serum for healthy people was in the range of 0–70 pg/ml (25.5 ± 13.8 pg/ml, n = 37). Secondary a relationship between the HPLC-ECD and ELISA methods was investigated. ELISA method could not detect 8-OHdG concentration in serum for healthy people, because the detection limit of 130 pg/ml was higher than the normal range for healthy people. These results show our SPE method has high sensitivity and quantitative accuracy for 8-OHdG analysis.
Keywords:Human serum  8-Hydroxy-2&prime  &ndash  deoxyguanosine (8-OHdG)  Solid phase extraction (SPE)  High performance liquid chromatography (HPLC)  Electrochemical detector (ECD)
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