HPLC法测定氯法拉滨有关物质及含量的方法学研究

Methodology research on determination of Clofarabine and its related substances by HPLC

目的建立有一种可以有效测定氯法拉滨有关物质与含量的高效液相色谱法，并进行质量方法学研究。方法色谱柱采用岛津C18柱（150mm×4．6mm，5μm），流动相为乙腈-水（10：90），检测波长为263nm，流速1．0mL·min^-1，柱温35℃，进样量20μL。结果氯法拉滨进样量在0．5028-4．0224μg范围内具有良好的线性关系（r：0．9995）；检测限（S／N=3）为2ng，定量限（S／N=10）为5ng。结论本方法准确，重现性好，可有效地检测氯法拉滨的有关物质和含量，并为氟法拉滨的检查及质量方法学研究提供了准确可靠的依据。

Objective To establish an HPLC method for determination of Clofarabine and its related substance, and to do research on quality methodology. Methods A Shimadzu C18 column （ 150 mm ×4.6mm,5μm） was adopted;the mobile phase was acetonitrile - water （ 10： 90） at a flow rate of 1.0 mL · min^-1 ;The detection wavelength was 263 nm;the temperature of the columnwas 35 ℃ and the injection volume was 20 μL. Results The linear range of Clofarabine was 0.502 8 -4.022 4 μg （ r = 0. 999 5 ） ; The limit of detection （ S/N = 3 ） was 2 ng and the limit of quantification （ S/N = 10 ） was 5 ng. Conclusion The method was accurate with good reproducibility, and was suitable for determination of Clofarabine and its related substances, and provided a reliable method for sample examination and methodology research of Clofarabine.