HPLC法测定异丙托溴铵原料药中的有关物质

目的:建立测定异丙托溴铵原料药中有关物质的方法。方法:采用高效液相色谱法。色谱柱为SHISEIDO C18,流动相为磷酸盐溶液(含0.1 mol/L磷酸二氢钠和0.01 mol/L四丙基氯化铵,pH 5.5)-甲醇(87∶13,V/V),流速为1 ml/min,检测波长为220nm,柱温为40Ⅴ,进样量为5μl。结果:异丙托溴铵和杂质B、C、D、E检测质量浓度分别在6.24⁶2.4、4.2562.4、4.25⁵3.1、4.5953.1、4.59⁵7.4、10.3557.4、10.35¹29.4、4.00129.4、4.00⁵0.01μg/ml范围内与各自峰面积积分值呈良好的线性关系(r=0.999 6、0.999 0、0.999 8、0.999 7、0.999 7);精密度、稳定性、重复性试验的RSD<10%;杂质B、C、D、E平均加样加收率分别为103.8%、104.3%、101.6%、104.4%,RSD分别为3.8%、2.2%、1.8%、3.5%(n=9)。结论:该方法准确、简便、快速,可用于异丙托溴铵原料药中有关物质的测定。

Determination of Related Substances in Ipratropium Bromide by HPLC

OBJECTIVE： To establish a method for the determination of related substances in ipratropium bromide. METHODS： HPLC method was adopted. The determination was performed on SHISEIDO C18 column with mobile phase consisted of phosphate solution （containing 0.1 mollL monosodium orthophosphate and 0.01 mol/L tetrapropylammonium chloride, pH 5.5）-methanol （87： 13, VIV） at the flow rate of 1 ml/min. The detection wavelength was set at 220 nm. The column temperature was 40 ℃ and sample size was 5μl. RESULTS： The linear range of ipratropium bromide and impurities B, C. D and E were 6.24-62.4 μg/ml （r=0.999 6）. 4.25-53.1 μg/ml （r=0.999 0）, 4.59-57.4 μg/ml （r=0.999 8）, 10.35-129.4 μg/ml （r=0.999 7） and 4.00-50.01 μg/ml （r=0.999 7）. respectively. RSDs of precision, stability and reproducibility tests were all lower than 10%. Average recoveries of impurities B, C, D and E were 103.8% （RSD=3.8%. n=9）, 104.3% （RSD=2.2%, n=9）, 101.6% （RSD=1.8%. 11=9） and 104.4% （RSD=3.5%. n=9）, respectively. CONCLUSIONS： The method is accurate, simple and rapid. and can be used for the determination of related substances in ipratropium bromide.