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目的建立人尿液中8-羟基脱氧鸟苷(8-OHd G)检测方法并探讨其在肿瘤诊断中的应用价值。方法以8-羟基脱氧鸟苷为外标,尿液经离心过滤后进行分析。色谱柱:Agilent HC-C18Column(4.6 mm×250 mm,5μm);流动相为50 mmol·L~(-1)Na H_2PO_4溶液(p H=3.5)-甲醇=90%∶10%,流速:1.0m L·min~(-1);检测波长:300 nm;进样量:50μL。用HPLC法检测肿瘤患者尿液中8-OHd G质量浓度,对照组为健康人,试验组纳入肺癌患者手术前后、肿瘤患者化疗前后的病例。结果该方法线性关系良好;最低定量限为0.1μg·m L~(-1);日内、日间精密度(RSD)均<2%;加标回收率均>90%。肺癌术前组、肺癌术后组与健康人组的8-OHd G水平分别为(5.49±5.48),(0.51±0.63),(1.16±1.65)μg·m L~(-1)。肺癌术前组的8-OHd G质量浓度显著高于肺癌术后组和健康人群组,差异有统计学意义(P<0.05)。结论用HPLC法检测尿液中的8-OHd G在肿瘤诊断和化疗疗效评估方面具有较高的准确度和特异性。
Objective To establish a method for the determination of 8-hydroxydeoxyguanosine (8-OHdG) in human urine and to explore its value in tumor diagnosis. Methods 8-hydroxy-2-deoxyguanosine as the external standard, urine after centrifugation analysis. Column: Agilent HC-C18 Column (4.6 mm × 250 mm, 5 μm); mobile phase was 50 mmol·L -1 Na 2 PO 4 solution (p H = 3.5) -methanol = 90% m L · min ~ (-1); detection wavelength: 300 nm; injection volume: 50 μL. The concentration of 8-OHd G in the urine of the cancer patients was detected by HPLC. The control group was healthy. The patients in the test group were included in the cases before and after the operation of lung cancer patients before and after chemotherapy. Results The method had a good linearity. The lowest limit of quantification was 0.1 μg · m L -1. The intra-day and inter-day RSD were all less than 2%, and the recoveries were both> 90%. The levels of 8-OHd G in lung cancer preoperative group, postoperative lung cancer group and healthy people were (5.49 ± 5.48), (0.51 ± 0.63) and (1.16 ± 1.65) μg · m L -1, respectively. The concentration of 8-OHd G in lung cancer preoperative group was significantly higher than that in postoperative lung cancer group and healthy group (P <0.05). Conclusion The HPLC method for the determination of 8-OHd G in urine has high accuracy and specificity in tumor diagnosis and chemotherapy efficacy evaluation.