HPLC法测定地西他滨中L-异构体和α-异构体含量

Determination of L-isomer and α-isomer in Decitabine by HPLC

目的建立高效液相法对地西他滨中L-异构体和α-异构体进行拆分并检测其中异构体含量。方法色谱柱采用CHIRACPAK-ADH手性柱(250mm×4.6mm,5μm);流动相:无水乙醇一正己烷(50∶50);流速:0.5m L·min^(-1);检测波长:244nm;进样量:20μL。结果本法对L-异构体和α-异构体的分离度为2.42,地西他滨浓度在0.395~4.02μg·m L^(-1)范围内呈良好的线性关系(r=0.9999),L-异构体浓度在0.419~4.04μg·m L^(-1)范围内呈良好的线性关系(r=0.9997),α-异构体浓度在0.419~4.02μg·m L^(-1)范围内呈良好的线性关系(r=0.9998),检测限分别为5.10ng和5.08ng,平均回收率分别为98.47%和99.22%。结论该方法操作简便,分离效果好,可用于地西他滨中异构体的含量测定。

OBJECTIVE To establish an HPLC method for separation of the L-and α-decitabine and determination of the L-and-α isomer. METHODS The separation was performed on a CHIRACPAK-ADH chiral column（ 250 mm × 4. 6mm,5μm）. Absolute alcohol and n-hexane（ 50∶ 50） was used as mobile phase. The flow rate was0. 5m L· min（-1）. The detection wavelength was set at 244 nm. The injection volume was 20μL. RESULTS This method showed that the resolution of the L-and-α isomer was 2. 42,the calibration curve was liner within the range of0. 395 - 4. 02μg· m L（-1）（ r = 0. 9999） for decitabine,the calibration curve was liner within the range of 0. 419 -4. 04μg·m L（-1）（ r = 0. 9997） for L-isomer,the calibration curve was liner within the range of 0. 419 - 4. 02μg·m L（-1）（ r = 0. 9998） for α-isomer,the L-isomer detection limit was 5. 10 ng,the α-isomer detection limit was 5. 08 ng,the average recovery rate was 98. 47% and 99. 22%. CONCLUSION The method is simple and effective. It can be used for content determination of L-and-α-isomer in decitabine.