高效液相色谱法测定人血清中假尿嘧啶核苷的浓度

Determination of Pseudourdine in Serum by HighPerformance Liquid Chromatography

应用反相高效液相色谱法测定人血清中假尿苷（ＰＤ）的含量，色谱柱为Ｎｏｖａ－ＰａｋＣ１８３．９ｍｍ×１５０ｍｍ，流动相为０．０４ｍｏｌ／Ｌ磷酸二氢钾缓冲液（ｐＨ４．０），检测波长为２６３ｎｍ，线性范围为０．７～６．８μｍｏｌ／Ｌ，回收率为９３．５０％，日间误差ＣＶ＝３．１１％（ｎ＝６）。同时测定了部队体检正常人血清中ＰＤ的浓度，并用于临床观察肝炎、肾病、肺癌等多种疾病以及Ｈｅ－Ｎｅ激光治疗前后患者血中ＰＤ含量的变化。正常人血中ＰＤ的浓度无性别差异，成年人的正常值与文献一致。肾脏疾病中以尿毒症患者血清中的假尿苷升高最为明显，３例肺癌患者也都有升高的趋势。而我院开展的Ｈｅ－Ｎｅ激光治疗前后无明显的变异，证明激光对细胞修饰没有显著的影响。

Pseudourdine is a modified nucleoside derived from the degradation of transfer fibonucleic acid. The elevation of modified nucleosides in urine has been suggested to be caused by higher turnover rate of t RNA in tumor tissue than in healthy tissue, rather than by cell death. A method of pseudourdine determination in serum was developed by high performance liquid chromatography on a Nova Pak column(Waters) with 0 04mol/L KH 2PO 4(pH 4 0)as mobile phase. The blood samples were collected and 0 6mL of serum was treated with 0 4mL 6% HClO 4. The precipitate was centrifuged for 10min at 3 000r/min. Five hundred μL of the liquor was dried by air stream at 60℃. The residue was dissolved with 300μL mobile phase and 10μL was injected. The average recovery was 93 50±2 1%. The calibration curve was linear within the concentration range of 0 7 6 8μmol/L. The serum pseudourdine concentrations for patients with hepatitis, lung cancer, hephritis and uremia were determined and those of patients with lung cancer and uremia were found significantly higher than those of healthy controls ( p <0 05). And for patients treated with He Ne lasser no significant change of the pseudourdine hasn ′t been found ( p <0 05).