反相高效液相色谱法同时测定人血浆5种嘌呤物质的含量

Simultaneous determination of five oxypurines in plasma by high-performance liquid chromatography

摘要:

目的：建立了高效液相色谱法同时测定血浆中5种嘌呤物质含量的方法，比较正常人与白血病和肝硬化患者血浆中5种嘌呤的含量差异，考察它们与白血病（Leukemia）和肝硬化（Liver cirrhosis）疾病的关系。方法：色谱柱：Phenomenex C18（250 mm×4.6 mm，5 μm；流动相：50 mmol/L醋酸铵（pH 4.65）-甲醇（99∶1，v/v）；流速：1 ml/min；检测波长：254 nm；柱温：30℃；进样量：100 μl。结果：尿酸在0.5~200 μg/ml浓度范围内线性关系良好，相关系数r=0.999 7；次黄嘌呤、腺嘌呤和黄嘌呤在0.005~0.3 μg/ml浓度范围内线性关系良好，相关系数r分别为0.999 8、0.999 3和0.999 9；鸟嘌呤在0.015~0.300 μg/ml浓度范围内线性关系良好，相关系数r=0.997 1。结论：白血病和肝硬化患者血浆中的尿酸、次黄嘌呤和黄嘌呤含量明显高于正常人，腺嘌呤和鸟嘌呤无明显差异。

Abstract:

Objective：To develop and validate a simple HPLC method to determine uric acid（UA），xanthine（Xan），hypoxanthine （Hyp），adenine（Ade） and guanine（Gua） in plasma of normals and the patients with Leukemia and liver cirrhosis. Methods：The separation was performed on Phenomenex C18 column（250 mm×4.6 mm，5 μm），with 50 mmol/L ammonium acetate（pH 4.65）-methanol（99∶1，v/v） as mobile phase isocratic elution，and measured absorbance at 254 nm. Results:Uric acid had good linear rela－tionship in the range of 0.5~200 μg/ml，and the correlation coefficient was 0.999 7；hypoxanthine，adenine and xanthine had good lin－ear relationships in the range of 0.005~0.300 μg/ml，and the correlation coefficients were higher than 0.999 3；guanine had good lin－ear relationships in the range of 0.015~0.300 μg/ml，and the correlation coefficient was 0.997 1. Conclusion：There are significant differences in the concentration of uric acid，hypoxanthine and xanthine in plasma between the momals and the patients with leukemia or liver cirrhosis，while adenine and guanine do not differ significantly.