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Improved method of plasma 8-Isoprostane measurement and association analyses with habitual drinking and smoking Soichi Kitano, Hisashi Hisatomi, Nozomu Hibi, Katsumi Kawano, Shoji Harada Soichi Kitano, Hisashi Hisatomi, Nozomu Hibi, Katsumi Kawano, Technology Development Department, SRL Inc., Hachioji, Tokyo, Japan Shoji Harada, Center for Molecular Biology and Cytogenetics, SRL Inc. , Hino, Tokyo, Japan Correspondence to: Soichi Kitano, Development Planning Section, Technology Development Department, SRL Inc., 5-6-50 Shinmachi, Hino-shi, Tokyo, 191-0002, Japan. firstname.lastname@example.org AIM: To develop a simple and accurate method for quantifying 8-isoprostane in plasma by employing a combination of two-step solid-phase extraction of samples and a commercially available ELISA kit, and by this method to examine the effects of drinking and smoking habits against the levels of plasma 8-isoprostane in healthy Japanese volunteers. METHODS: Plasma 8-isoprostane was extracted with ODS gel suspension followed by NH2 Sep-Pak column. The 8-isoprostane fractions were assayed using a commercially available ELISA kit. We measured plasma 8-isoprostane levels in 157 healthy Japanese volunteers divided into three groups (64 non-habitual drinkers, 56 moderate drinkers and 37 habitual drinkers) according to their alcohol consumption per week. Genotypes of aldehyde dehydrogenase 2 (ALDH2) were also determined to investigate the plasma 8-isoprostane levels with reference to drinking habits. In addition, the plasma 8-isoprostane levels of 96 non-smokers and 61 smokers from the same subjects were compared. RESULTS: Our method fulfilled all the requirements for use in routine clinical assays with respect to sensitivity, intra- and inter-assay reproducibility, accuracy and dynamic assay range. Significant increases of plasma 8-isoprostane levels were observed in female habitual drinkers when compared with those of non-habitual drinkers (t = 5.494, P CONCLUSION: Our present method was proved to be a simple and accurate tool for measuring plasma 8-isoprostane. However, the clinical utility of plasma 8-isoprostane for drinking and smoking habits was limited since elevated 8-isoprostane levels were observed in female heavy drinkers, and no association was found between smokers and nonsmokers. Key words: 8-Isoprostane; ELISA; Lipid peroxidation; Drinking; Smoking
Source: World J Gastroenterol 2006 September 28;12(36):5846-5852.
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