Isolation and identification of urinary nucleosides. Applications of high-performance liquid chromatographic methods to the synthesis of 5'-deoxyxanthosine and the simultaneous determination of 5,6-dihydrouridine and pseudouridine.
Isolation and identification of urinary nucleosides. Applications of high-performance liquid chromatographic methods to the synthesis of 5'-deoxyxanthosine and the simultaneous determination of 5,6-dihydrouridine and pseudouridine.
复制标题
尿核苷的分离和鉴定。
DOI：
10.1016/s0021-9673(01)89352-3
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发表时间：
1990
期刊：
影响因子：
--
通讯作者：
Yamamoto,H
中科院分区：
文献类型：
--
作者：
Nakano,K;Yasaka,T;Schram,KH;Reimer,ML;McClure,TD;Nakao,T;Yamamoto,H
用硼酸盐亲和凝胶柱从正常人尿液中提取修饰核苷，并用反相高效液相色谱(RP-HPLC)进行分离。用三甲基硅烷基衍生物气相色谱-质谱法测定了30个反相高效液相色谱组分中的主要成分。以5‘-脱氧腺苷为原料，采用反相高效液相色谱法合成5’-脱氧黄嘌呤。此外，采用串联的两个ODS柱，用反相高效液相色谱法同时测定了尿中5，6-二氢尿苷(D)和假尿苷(Ψ)。正常混合尿中D水平为4.87nmol/μ肌酐。用反相高效液相色谱法测定了4例胃肠道肿瘤患者手术前后尿液中D和Ψ的含量。在四名癌症患者中，有三名患者在手术后尿液中这两种核苷水平都有较大幅度的下降。
Modified nucleosides from pooled normal human urine were extracted using a boronate affinity gel column and fractionated by reversed-phase high-performance liquid chromatography (RP-HPLC). The major constituents in each of the 30 RP-HPLC fractions were determined by gas chromatography-mass spectrometry of the trimethylsilyl derivatives of the fractions. The same RP-HPLC method was used in the synthesis of 5′-deoxyxanthosine from authentic 5′-deoxyadenosine. In addition, the simultaneous determination of urinary 5,6-dihydrouridine (D) and pseudouridine (Ψ) was carried out by RP-HPLC using two ODS columns in series. The level of D in pooled normal urine was 4.87 nmol/μmol creatinine. The RP-HPLC method was applied to the measurement of D and Ψ levels in urine collected before and after surgery from four patients with gastrointestinal cancer. A large decline in both nucleoside levels in urines after surgery was observed in three of the four cancer patients.