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目的 :应用高效液相色谱法对卡介苗素中游离核苷酸进行分离及含量测定。方法 :采用C18柱 ,以甲醇 -0 0 5mol·L-1硫氢酸四丁基铵 (用氨试液调节pH为 3 9) (2 8∶72 )为流动相 ,检测波长 2 6 0nm。结果 :卡介苗素中主要含有一磷胞苷、一磷尿苷、一磷鸟苷、三磷鸟苷及三磷腺苷 5种游离核苷酸 ,含量在 0 33%～ 1 5 8%之间 ,方法平均回收率为 96 71%～ 10 1 3% ,相对标准偏差为 1 1%～ 1 9%。结论 :该方法简便、快速、准确 ,专属性好 ,可用于测定样品中核苷酸的含量。
OBJECTIVE: To isolate and determine free nucleotides in BCG by high performance liquid chromatography. Methods: The mobile phase was C18 column with tetramethylammonium hydrogen sulphate of 0.05-0.50 mol·L-1 (pH adjusted to 30 with ammonia solution) (2 8:72). The detection wavelength was 260 nm. Results: BCG mainly contained five kinds of free nucleotides, monophosphatidylglycine, monophosphatidyluridine, monophosphatidylinosine, triphosphatidylinosine and adenosine triphosphate, with the content ranging from 0 33% to 158% The average recoveries were 96 71% -10 10% with relative standard deviations of 11% ~ 19%. Conclusion: The method is simple, rapid, accurate and specific and can be used to determine the nucleotide content in samples.