Tagesson C, Källberg M, Klintenberg C, Starkhammar H
Department of Occupational and Environmental Medicine, Faculty of Health Sciences, Linköping University, Sweden.
Eur J Cancer. 1995 Jun;31A(6):934-40. doi: 10.1016/0959-8049(94)00490-0.
An automated analytical method has been developed for determination of the oxidative DNA adduct, 8-hydroxydeoxyguanosine (8OHdG) in human urine, based on coupled-column high performance liquid chromatography with electrochemical detection. Urine is concentrated on Bondelut CH by means of an automated sample processor, and the enriched sample injected on to a polymeric reversed phase column coupled in line with an electrochemical detector and a C18 reversed phase column. By use of the electrochemical detector, a suitable retention time interval is set for collection of the fraction containing 8OHdG from the chromatography on the first column; this fraction is collected in a 2 mL loop and injected onto the C18 column. The system is operated by an automatic valve station controlled by an integrator. The method has a large sample capacity and measures 31.1, 15.7, and 7.43 nmol 8OHdG/L urine with variation coefficients of 8, 8 and 24% within series and 8, 11 and 23% between series. Normal healthy individuals were found to excrete 14.9 +/- 7.8 nmol 8OHdG/24 h, or 1.11 +/- 0.62 mumol 8OHdG per mol creatinine, in their urine, whereas increased levels of 8OHdG were found in 24 h collections from a variety of cancer patients, both in samples taken before onset of oncological therapy (1.84 +/- 1.12 mumol/mol creatinine, P < 0.01 versus healthy individuals) and after therapy onset (2.18 +/- 1.44 mumol/mol creatinine, P < 0.001 versus healthy individuals). Moreover, mean values of 8OHdG in random urinary samples from cancer patients were significantly higher than from healthy individuals (2.42 +/- 2.28 versus 1.19 +/- 0.48 mumol/mol creatinine, P < 0.001), both in samples taken before therapy onset (1.91 +/- 0.96, P < 0.001 versus healthy individuals) and after (2.57 +/- 2.46, P < 0.001 versus healthy individuals). High levels of urinary 8OHdG were found in patients subjected to whole body irradiation, and in patients receiving chemotherapy with various cytostatic agents. The potential use of the method for detecting increased urinary 8OHdG excretion and conditions associated with increased oxidative DNA damage in humans is discussed.
已开发出一种自动化分析方法，用于基于柱耦合高效液相色谱 - 电化学检测法测定人尿中的氧化性DNA加合物8 - 羟基脱氧鸟苷（8OHdG）。尿液通过自动进样器在Bondelut CH上进行浓缩，然后将富集后的样品注入到与电化学检测器和C18反相柱在线耦合的聚合物反相柱上。通过使用电化学检测器，设定合适的保留时间间隔，以便从第一根柱的色谱图中收集含有8OHdG的馏分；该馏分收集到一个2 mL定量环中，然后注入到C18柱上。该系统由积分仪控制的自动阀站操作。该方法具有较大的样品容量，对尿中8OHdG的测定值为31.1、15.7和7.43 nmol/L，同组内变异系数分别为8%、8%和24%，不同组间变异系数分别为8%、11%和23%。发现正常健康个体尿中8OHdG的排泄量为14.9±7.8 nmol/24 h，即每摩尔肌酐中含1.11±0.62 μmol 8OHdG，而在各类癌症患者的24小时尿液收集样本中，8OHdG水平升高，无论是在肿瘤治疗开始前采集的样本（1.84±1.12 μmol/mol肌酐，与健康个体相比P<0.01）还是治疗开始后（2.18±1.44 μmol/mol肌酐，与健康个体相比P<0.001）。此外，癌症患者随机尿样中8OHdG的平均值显著高于健康个体（2.42±2.28与1.19±0.48 μmol/mol肌酐，P<0.001），无论是在治疗开始前采集的样本（1.91±0.96，与健康个体相比P<0.001）还是治疗后（2.57±2.46，与健康个体相比P<0.001）。在接受全身照射的患者以及接受各种细胞抑制剂化疗的患者中发现尿中8OHdG水平较高。本文讨论了该方法在检测人尿中8OHdG排泄增加及与氧化性DNA损伤增加相关情况方面的潜在用途。
