基于HPLC法的人尿液8-羟基脱氧鸟苷检测在临床肿瘤诊断的应用  

Application of human urine 8-hydroxy deoxyguanosine detection by HPLC method in clinical tumor diagnosis

作　　者：于晨[1,2] 董超然[1,2] 康晓征[3] 张照辉[4] 黄健[1,2] 曾慧慧[1,2] 

机构地区：[1]北京大学天然药物及仿生药物国家重点实验室,北京100191 [2]北京大学药学院化学生物学系,北京100191 [3]北京大学肿瘤医院暨恶性肿瘤发病机制及转化研究教育部重点实验室,北京100142 [4]北京大学第三医院肿瘤化疗与放射病科,北京100191

出　　处：《中国临床药理学杂志》2017年第17期1701-1705,共5页The Chinese Journal of Clinical Pharmacology

基　　金：国家自然科学基金资助项目(81372266)

摘　　要：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 develop a method for the detection of 8 - hy- droxy deoxyguanosine （8 - OHdG） in urine and to explore its value in tumor- assisted diagnosis. Methods The chromatographic separation was performed on Agilent HC - CxsColumn （4.6 mm x 250 ram,5 Ixm） with the mobile phase of 90% 50 mmol · L-1 NaH2PO4aqueous solution and 10% methanol at a flow rate of 1.0 mL · min-1 The detection wavelength was 300 nm. The injection volume was 50 μL. The concentra- tion of 8 - OHdG in the urine of patients with cancer was detected by HPLC. The control group was healthy person and the patients were divided into two groups：before and after the operation, before and after chem- otherapy. Results The method had a good linear relationship. The limit of detection was 0. 1 μg·m L-（-1）. The RSDs of inter - day and intra - day precision were less than 2%. The spiked recovery rates were more than 90%. The level of 8 - OHdG in lung cancer group was （5.49 ± 5.48 ） μg·m L-（-1）, the level of 8 -OHdG in lung cancer group was （0. 51 ±0. 63 ）μg·m L-（-1）, the level of 8 - OHdG in healthy group was （1.16 ± 1.65 ）μg·m L-（-1） The concentration of 8 -OHdG in preopretive group was significantly higher than that in postoperative groupand control group （ all P 〈 0. 05 ）. Conclusion 8 - OHdG detected by HPLC has high accuracy and specificity in tumor diagnosis and evaluation of chemotherapy efficacy.

关 键 词：8-羟基脱氧鸟苷 肿瘤标志物 高效液相色谱法 

分 类 号：R979.1[医药卫生—药品]