RP-HPLC法测定注射用尖吻蝮蛇血凝酶的含量

目的: 建立测定注射用尖吻蝮蛇血凝酶的高效液相色谱分析方法。方法: 采用Vydac 208TP54-C₈(5 μm,4.6 mm×250 mm)色谱柱,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),梯度洗脱,流速为0.8 mL·min^-1,柱温45 ℃,检测波长为280 nm。结果: 尖吻蝮蛇血凝酶浓度在2.5～30 μg·mL^-1(r=0.9991)范围内线性关系良好,平均回收率(n=9)为99.5%。结论: 该方法可以为注射用尖吻蝮蛇血凝酶的质量控制提供依据。

Objective: To establish an HPLC method for the determination of heamocoagulase agkistrodon for injection. Methods: The separation was performed on Vydac 208TP54-C₈ (5 μm, 4.6 mm×250 mm) column with the mobile phase A of 0.1% trifluoroacetic acid and B of 90% acetonitrile and water(90: 10, containing 0.1% trifluoroacetic acid), at a flow rate of 0.8 mL·min^-1, with gradient elution.The column temperature was 45 ℃, and the detection wavelength was set at 280 nm. Results: The linear range was 2.5-30 μg·mL^-1 (r=0.9991) for heamocoagulase agkistrodon.The average recovery (n=9) was 99.5%. Conclusion: The method can provide a scientific basis for the quality control of heamocoagulase agkistrodon for injection.

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