【摘要】 利用高效液相色谱方法在普通Polar-RP柱上成功分离pentostatin的4个异构体。通过考察色谱柱、流动相、温度等对分离度的影响,得到最佳分离条件为:室温下以Phenom enex Synergi Polar-RP色谱柱,采用磷酸氢二钠水溶液(50 mmol,pH 8.8)-甲醇(97∶3,V/V)为流动相;282 nm检测;流速为1.0 mL/m in。结果表明,4个异构体之间均达到基线分离,且保留时间和峰面积的RSD均控制在0.6%以内。pentostatin在0.1~1.0g/L、其异构体在0.5~1.5 mg/L范围内的线性相关系数(R)均大于0.99。在此实验基础上,建立分离纯化Pentostatin的制备色谱条件,以制备色谱法得到的产品纯度达到99.8%,收率达到理论产率的80.2%。更多还原

【Abstract】 Pentostatin and its isomers were separated by high performance liquid chromatography(HPLC)(using) Polar-RP column.The influence of column,mobile phase,and temperature was investigated to obtain the optimal chromatographic condition.Phenomenex Synergi Polar-RP column was used at room temperature.The mobile phase was bisodium phosphate buffer(50 mmol,pH 8.8)-methanol in the ratio of 97∶3(V/V).Flow rate was kept at 1.0 mL/min and the elution was monitored at 282 nm.Pentostatin and its(isomers) were baseline separated and the retention time and peak area reproducibility of four isomers(RSD) were less than 0.6% respectively.The correlation coefficients(R) of pentostatin(0.1-1.0 g/L) and(isomers)(0.5-1.5 mg/L) were over 0.99.Based on the analytical method,a preparative-LC method was developed to separate pentostatin and its isomers.pentostatin product by Pre-LC purification has 99.8% purity and 80.2% yield.更多还原