HPLC梯度洗脱法测定复合物中特考韦瑞和烟酰胺的含量

Determination of content for tecovirimat and nicotinamide in the complex by HPLC with gradient elution method

目的建立HPLC梯度洗脱法同时测定特考韦瑞-烟酰胺复合物中特考韦瑞和烟酰胺的含量。方法采用Waters Symmetry C18色谱柱(4.6 mm×250 mm,5μm),以50 mmol/L磷酸二氢钾溶液-乙腈为流动相梯度洗脱,流速为1.0 ml/min,柱温为30℃,检测波长为224 nm,进样量为20μl。结果特考韦瑞和烟酰胺之间分离度良好;特考韦瑞在5~50μg/ml、烟酰胺在5~50μg/ml范围内峰面积与浓度线性关系良好;其平均回收率分别为100.76%和100.01%;该法的重复性及中间精密度均符合要求;检测用溶液在室温条件下放置24 h稳定。结论该法准确可靠,可用于复合物中特考韦瑞和烟酰胺含量的同时测定。

Objective To establish an HPLC method to determine the concentration of tecovirimat and nicotinamide in the tecovirimat/nicotinamide complex. Methods A Waters Symmetry C18 column（ 4. 6 mm × 250 mm,5 μm） was used with 50 mmol/L KH2 PO4-acetonitrile as the mobile phase with gradient elution at a flow rate of 1. 0 ml/min. The column temperature was 30℃ and the detection wavelength was 224 nm. The injection volume was 20 μl. Results Tecovirimat and nicotinamide were separated completely. The calibration curves were linear within the range of 5 ~ 50 μg/ml for tecovirimat and 5 ~ 50 μg/ml for nicotinamide. The average recoveries were 100. 76% and 100. 01%,respectively. The repeatability and intermediate precision met the requirements. The test solution was stable within 24 h. Conclusion This method is reliable,so it can be used for the simultaneous determination of tecovirimat and nicotinamide in the complex.