整体柱高效液相色谱法测定尿中核苷的方法研究

High Performance Liquid Chromatographic Method Development Using Monolithic Column for the Determination of Urinary Nucleosides

尿中修饰核苷已被研究用作可能的肿瘤标记物,目前最常用的分析方法为反相高效液相色谱法,其不足之处是分析周期太长。为此,建立了应用整体柱的高效液相色谱方法对尿中修饰核苷进行分析,所用缓冲液为25mmol/L磷酸二氢钾溶液(pH4 55)和60%(体积分数)的甲醇水溶液,线性梯度洗脱,检测波长为260nm。12种目标核苷可被完全分离。所建立的方法在分离度、线性、重现性、灵敏度及回收率等指标上与普通反相液相色谱法相近,但分析周期大大缩短,仅为23min,适合于大量临床样品的分析。

Modified nucleosides excreted in urine have been studied as possible bio-markers for malignant tumors. Reversed-phase high performance liquid chromatography (RP-HPLC)is one of the most popular analytical tools. However, its analysis time is longer than desired. The (analytical) method for urinary nucleosides has been developed using a monolithic column and a (linear) gradient elution of 25 (mmol/L) KH_2PO_4 solution (pH 4.55) and 60% methanol in water with UV detection at 260 nm. Twelve urinary nucleosides were completely separated. The monolithic column presented similar analytical characteristics to reversed-phase column. The time of the analysis is only 23 min, in general much shorter than the methods using a packed RP-HPLC. The method developed is sensitive, reliable and suitable for clinical applications.