高效液相色谱法测定人血红细胞内硫鸟嘌呤核苷酸浓度

目的:建立以高效液相色谱法测定人血红细胞内硫鸟嘌呤核苷酸浓度的方法。方法:色谱柱为HypersilODS,流动相为甲醇-水(10∶90),检测波长为342nm;硫鸟嘌呤核苷酸经加热水解生成6-硫鸟嘌呤(6-TG),6-TG经提取至0.1mol/L盐酸中进样分析,以外标法定量。结果:6-TG浓度线性范围为30～1200pmol/8×108RBCs,水解时间以1h为宜,提取时pH值以11～12为宜,提取液中醋酸苯汞的量为1.3mmol/L。结论:在优化条件下,本方法测定硫鸟嘌呤核苷酸快速、准确、灵敏度高。

Determination of Thioguanosine in Human Red Blood Cells by HPLC

OBJECTIVE: To establish HPLC method for the determination of thioguanosine in human red blood cells. METHODS: The determination was performed on Hypersil ODS colum. The mobile phase consisted of methanol-water (10∶90) with detection wavelength at 342nm. 6-Thioguanine(6-TG)was generated from thioguanosine through heating and hydrolyzing. 6-TG was extracted into 0.1mol/L hydrochloric acid for sample injection assay with external reference method for the quantification. RESULTS: The linear range of 6-TG was 30～1 200pmol/8×108RBCs. Its optimum hydrolyzing time was 1 hour and optimal extraction pH value ranged from 11 to 12. The content of phenyl mercury acetate in the extraction solution was 1.3mmol/L. CONCLUSION: Under optimized conditions, HPLC method for the determination of thioguanosine is fast, accurate and sensitive.