RP-HPLC法测定注射用降纤酶中降纤酶的含量

RP-HPLC determination of defibrase in defibrase for injection

目的:建立测定注射用降纤酶中降纤酶的高效液相色谱分析方法。方法:色谱柱为Vydac 214TP54-C4(5μm,250 mm×4.6 mm),流动相A为0.1%三氟乙酸,B为乙腈-水(90:10,含0.1%三氟乙酸),流速为0.8 mL.min-1,采用梯度洗脱,柱温45℃,检测波长为280 nm。结果:降纤酶在0.5～32μg.mL-1(r=0.9999)范围内线性关系良好,平均回收率(n=9)为98.5%。结论:该法快速、灵敏、准确,可以为注射用降纤酶的质量控制提供依据。

Objective：To establish an HPLC method for the determination of defibrase in defibrase for injection.Method：The separation was performed on Vydac 214TP54-C4（5 μm,250 mm×4.6 mm）column with the mobile phase of A 0.1% trifluoroacetic acid and B 90%acetonitrile（containing 0.1% trifluoroacetic acid）,at flow rate of 0.8 mL·min-1,gradient elution adopted.The column temperature was 45 ℃,and the detection wavelength was set at 280 nm.Results：The linear ranges were 0.5-32 μg·mL-1（r=0.9999）for defibrase.The average recovery（n=9） was 98.5%.Conclusion：A rapid,sensitive and accurate method can provide a scientific basis for the quality control of defibrase for injection.