高效液相色谱法测定注射用氟脲苷的含量及有关物质

HPLC determination of floxuridine content and its related substances in floxuridine solution for injection

目的：建立同时测定注射用氟脲苷含量及有关物质的HPLC方法。方法：采用反相高效液相色谱法，色谱柱为Venusil XBP C18（4.6 mm×250 mm，5 μm），以磷酸盐缓冲液（用磷酸调pH至6.6）为流动相A，流动相A-甲醇（50：50）为流动相B，梯度洗脱：（0-20 min，95% A→60% A；20-30 min，60% A；30-35 min，60% A→95% A），流速为1.0 mL·min^-1；柱温：35 ℃；检测波长为268 nm。结果：氟脲苷峰与各主要杂质峰均分离良好，氟脲苷浓度在63.48-571.32 μg·mL^-1范围内，与峰面积呈良好的线性关系，回归方程A=15.822C＋13.224，r=1.000（n=7）；重复性试验RSD为0.41%（n=6）；平均加样回收率（n=9）为99.2%（RSD=0.48%）；最低检测限为21.08 ng；供试品溶液在24 h内稳定。结论：本方法准确、灵敏，专属性强，适用于注射用氟脲苷的质量控制。

Objective： To establish an HPLC method to determine the floxuridine content and its related substances in the floxuridine solution for injection. Methods： The analysis was achieved on a Venusil XBP C18 column （250 mm×4.6mm,5 μm） using a mobile phase of methonal-phosphoric acid buffer（pH adjusted to 6.6 with phosphoric acid） with gradient elution in a total runing time of 35 min（0-20 min,95% A→60% A;20-30 min,60% A;30-35 min,60% A→95% A）.The flow rate was 1.0 mL·min^-1 and the column temperature was 35 ℃ and the detection wave length was 268 nm. Results： There was a good resolution among the peaks of floxuridine and the main related substances.There was a good linear relationship between the peak area and the concentration of floxuridine in the range of 63.48-571.32 μg· mL^-1 （r=1.000,n=7）.Regression equation could be expressed by A=15.822C＋13.224.The average recovery was 99.2%（RSD=0.48%,n=9）,the RSD of the repeatability test was 0.41% （n=9）,and the limit of detection was 21.08 ng.The solution was stable for 24 hours. Conclusions： The method is specific,accurate and sensitive,which can be used for quality control of the floxuridine solution for injection.