Quantitation of urinary nucleosides by high-performance liquid chromatography
Quantitation of urinary nucleosides by high-performance liquid chromatography
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DOI：
10.1016/s0021-9673(96)00757-1
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发表时间：
1997-02-28
影响因子：
4.1
通讯作者：
Schmid, HR
中科院分区：
文献类型：
--
作者：
Liebich, HM;DiStefano, C;Schmid, HR
已知一些源自RNA降解的修饰的，特别是甲基化的核苷在恶性肿瘤患者的尿液中以异常水平排泄，并且它们已被提议作为肿瘤标志物。它们的测量可以提供一种非侵入性的诊断方法，有助于识别不同的癌症和监测治疗效果。在这项研究中，我们开发并优化了一种分析方法，以分离和定量正常和修饰的核糖核苷。尿中核苷的提取是通过在苯硼酸柱上进行亲和层析分离之前进行的。反相高效液相色谱法允许一个完整的分离16尿核糖核苷。不同核苷的回收率范围为83 - 100%，除了黄嘌呤核苷（66%）和假尿苷（74%）。在正常24小时尿中，13种核苷的平均水平（以nmol核苷/μ mol肌酐计）如下：二氢尿苷（6.37），假尿苷（25.52），胞苷（0.07），尿苷（0.21），1-甲基腺苷（2.19），肌苷（0.30），鸟苷（0.06），黄嘌呤核苷（0.59）、3-甲基尿苷（0.11）、1-甲基肌苷（1.13）、1-甲基鸟苷（0.74）、腺苷（0.21）和5 ′-脱氧-5 ′-甲硫基腺苷（0.12）。关于两种肿瘤，即乳房肿瘤和口底肿瘤的初步结果显示，核糖核苷的含量有些异常。现在正在进行进一步的实验，以测量不同形式的癌症患者尿液中的修饰核苷。
It is known that some modified, especially methylated, nucleosides originating from RNA degradation are excreted in abnormal levels in the urine of patients with malignant tumours and they have been proposed as tumour markers. Their measurement could provide a non-invasive diagnostic method, be helpful in the identification of different cancers and in the monitoring of therapeutic effects. In this study, we developed and optimized an analytical procedure to isolate and quantify normal and modified ribonucleosides. The extraction of urinary nucleosides was performed by affinity chromatography on a phenylboronic acid column prior to separation. The reversed-phase high-performance liquid chromatography method allowed a complete separation of sixteen urinary ribonucleosides. The recoveries for the different nucleosides ranged from 83 to 100%, except for xanthosine (66%) and pseudouridine (74%). In normal 24 h urine, the mean levels of thirteen nucleosides (in nmol of nucleoside/mu mol of creatinine) were found to be as follows: dihydrouridine (6.37), pseudouridine (25.52), cytidine (0.07), uridine (0.21), 1-methyladenosine (2.19), inosine (0.30), guanosine (0.06), xanthosine (0.59), 3-methyluridine (0.11), 1-methylinosine (1.13), 1-methylguanosine (0.74), adenosine (0.21) and 5'-deoxy-5'-methylthioadenosine (0.12). The first results concerning two kinds of tumours, i.e. breast and floor of mouth tumours, showed some abnormal levels of ribonucleosides. Further experiments are now in progress to measure the modified nucleosides in urine of patients with different forms of cancer.