HPLC同时测定慢性阻塞性肺病患者血浆中的茶碱和1,3-二甲基尿酸

Simultaneous determination of theophylline and 1,3-dimethyluric acid in the patients plasma with chronic obstructive pulmonary disease by HPLC

目的采用HPLC法同时测定慢性阻塞性肺病（COPD）患者血浆中的茶碱及其代谢物1,3-二甲基尿酸（1,3-DMU）。方法色谱柱为Hypersil BDS C18柱（250 mm×4.6 mm,5μm）,流动相为乙腈-水（7∶93）,流速1 m L·min-1,检测波长280 nm。43名COPD患者连续服用茶碱（100 mg,bid）两周后,测定茶碱及1,3-DMU的血药浓度。结果茶碱、1,3-DMU的线性范围分别为0.20~10.00μg·m L-1（r=0.9998）、0.05~2.00μg·m L-1（r=0.9996）;日内、日间RSD均≤12.91%,提取回收率分别为77.52%~79.22%、71.69%~74.02%;患者的平均茶碱血药浓度为3.57±1.59μg·m L-1,茶碱和1,3-DMU的浓度呈强相关（r=0.672）,1,3-DMU/茶碱为0.064±0.024。结论所用方法简便、快速、准确,可用于COPD患者茶碱血药浓度的监测,1,3-DMU与茶碱浓度的比值可评估不同个体对茶碱清除率的差异。

OBJECTIVE To establish an HPLC method for the simultaneous determination of plasma levels of theophylline and its metabolite 1,3-dimethyluric acid（ 1,3-DMU）. METHODS The HPLC analysis was performed on an Hypersil BDS C18column（ 250 mm × 4. 6 mm,5 μm） with a mobile phase consisting of acetonitrile and water（ 7∶93）. The flow rate was 1 m L·min-1and the detection wavelength was 280 nm. Plasma drug concentration in 43 patients medicated with theophylline（ 100 mg,bid） for two weeks was measured. RESULTS The linear ranges of theophylline and 1,3-DMU were 0. 20-10. 00 μg·m L-1（ r = 0. 9998） and 0. 05-2. 00 μg·m L-1（ r = 0. 9996）,respectively. The intra-and inter-day RSD were all less than 12. 91%. The extraction recovery was77. 52%-79. 22% for theophylline and 71. 69%-74. 02% for 1,3-DMU. The average plasma theophylline level of the 43 patients was 3. 57 ± 1. 59 μg·m L-1. A strong correlation was noted between the plasma theophylline and 1,3-DMU levels（ r = 0. 672）,and DMU / theophylline was 0. 064 ± 0. 024. CONCLUSION This method is simple,fast and accurate for simultaneous determination of human plasma levels of theophylline and 1,3-DMU,and is useful to assess the metabolic ability of individual patients.