肠癌患者尿中核苷排放的高效液相色谱法研究

Investigation of Urinary Nucleosides Excretion of Intestinal Cancer Patients by Reversed-Phase High Performance Liquid Chromatography

用反相高效液相色谱法测定尿中核苷。通过苯基硼酸亲和色谱法提取尿中核苷,在色谱柱(4 6mmi d ×250mm,5μm)上以25mmol/L磷酸二氢钾溶液(pH4 55)和60%的甲醇水溶液作为流动相进行二元梯度淋洗,于22℃下进行反相色谱分离,260nm处紫外检测。用该法测定了41例肠癌患者和52例正常人尿中15种核苷的含量(用核苷与肌酐的摩尔比表示,下同),结果表明肠癌患者中有12种核苷的含量比正常人显著性增高(P<0 001)。以15种核苷的含量作为参量,结合主成分分析区分正常人和肠癌患者,对癌症病人的识别率达76%(31/41)。该方法灵敏、可靠,适合恶性肿瘤防治的基础研究和临床应用。

A method for the determination of urinary nucleosides with reversedphase high performance liquid chromatography is described. After nucleosides were extracted from urine on phenyl boronic acid affinity chromatography, the analysis was performed on a column (46 mm id×250 mm, 5 μm) at 22 ℃ using a linear gradient elution comprising 25 mmol/L KH2PO4 solution (pH 455) and 60% methanol in water with UV detection at 260 nm. This method was used for the determination of 15 urinary nucleosides of 41 intestinal cancer patients and 52 normal adults. The results showed that the average concentrations of 12 urinary nucleosides from intestinal cancer patients were much higher than those of normal adults with P<0001. Using the concentrations of 15 nucleosides as the data vectors, principal component analysis was applied to classify intestinal cancer patients and normal adults and 76% (31/41) of the cancer patients were correctly classified. It is concluded that the method is sensitive, reliable and suitable for basic research and clinical applications to malignant tumours.