HPLC法测定富马酸沃诺拉赞原料药中吡啶-3-磺酸的含量

Determination of Pyridine-3-sulfonic in Vonoprazan Fumarate by HPLC

目的建立测定富马酸沃诺拉赞原料药中吡啶-3-磺酸的高效液相色谱(HPLC)法。方法色谱柱为Inertsil ODS-3 C18柱(4.6 mm×150 mm,5μm),以0.025 mol·L-1磷酸钠溶液(用磷酸调节p H值至6.8)-乙腈(98∶2,V/V)为流动相A,乙腈为流动相B,进行线性梯度洗脱,检测波长为210 nm,柱温为25℃,流速1.0 m L·min^(-1),进样量20μL。结果在选定的色谱条件下,吡啶-3-磺酸与相邻色谱峰分离良好,在0.197 4～1.974 3μg·mL^(-1)浓度范围内与峰面积呈良好的线性关系,线性方程为Y=41 638X+1 137.5,相关系数r=0.999 8;平均回收率为103.6%,RSD%为1.34%(n=12); 6批样品中吡啶-3-磺酸的含量分别为0.001 8%、0.001 7%、0.001 5%、0.001%、0.004%及0.004%。结论本方法精密度良好,灵敏度高,准确度好,适用于富马酸沃诺拉赞原料药中吡啶-3-磺酸的检测。

OBJECTIVE To establish an HPLC method for the determination of pyridine-3-sulfonic acid in vonoprazan fumarate.METHODS Inertsil ODS-3 C18( 4.6 mm×150 mm,5 μm) column was used,0.025 mol·L-1 sodium phosphate solution( pH adjusted to 6.8 with phosphori acid)-acetonitrile( 98 ∶2,V/V) as mobile phase A,acetonitrile as mobile phase B,using linear gradient elution.The detection wavelength was 210 nm,column temperature was 25 ℃,flow rate was 1.0 m L·min-1 and the sample volume was 20 μL.RESULTS Under the selected chromatographic conditions,pyridine-3-sulfonic acid was separated from adjacent chromatographic peaks,the concentration in the range of 0.197 4-1.974 3 μg·mL-1 had good linear relationship with the peak area,the linear equation was Y = 41 638 X+1 137.5,the correlation coefficient was r = 0.999 8. The average recovery rate was 103.6% and RSD% was 1.34%( n= 12).The content of pyridine-3-sulfonic acid in 6 batches of samples were 0.001 8%,0.001 7%,0.001 5%,0.001%,0.004% and0.004% respectively. CONCLUSION The method has been verified by methodology,and the method has good precision,high sensitivity and good accuracy. It is suitable for the detection of pyridine-3-sulfonic acid in the vonoprazan fumarate.