高效液相色谱法测定杯(6)芳烃磺酸钠及其有关物质

Determination of content and related substances of 6-sulfocalix (6) arene

目的：建立测定杯（6）芳烃磺酸钠的含量及有关物质的高效液相色谱方法。方法：采用SHISEIDO CAPCELL ADME S5（250 mm×4.6 mm,5.0μm）色谱柱;流动相A为乙腈,流动相B为10 mmol·L-1磷酸氢二铵＋10 mmol·L-1磷酸二氢铵,p H 7.0;梯度洗脱（0～30 min,2%A→21.5%A;30～35 min,21.5%A→2%A）;流速：1.0 m L·min-1,检测波长：208 nm;柱温：35℃;进样体积：20μL。结果：在上述色谱条件下,各杂质及各降解产物均可与杯（6）芳烃磺酸钠主峰良好分离,主要有关物质杯（4）芳烃磺酸钠与主峰分离良好;HPLC测定的线性范围为0.1～2μg（r=0.999 0）,方法的仪器精密度、日内精密度、日间精密度及重复性的RSD均小于2%;3批杯（6）芳烃磺酸钠原料药中杯（6）芳烃磺酸钠含量范围为98.14%～98.65%之间,总有关物质含量不高于1.0%。结论：经方法学验证,本法可用于测定杯（6）芳烃磺酸钠的含量及有关物质。

Objective：To establish an HPLC method for determination of the content and the related substances of 6-sulfocalix（6）arene.Methods：The SHISEIDO CAPCELL ADME S5（250 mm×4.6 mm,5 μm）column was adopted.The mobile phase A consisted of acetonitrile,and the mobile phase B consisted of 10 mmol·L-1（NH4）2HPO4＋10 mmol·L-1 NH4H2PO4,p H=7.0;the gradient elute was adopted（0-30 min,2%A → 21.5%A;30-35 min,21.5%A → 2%A）,and the flow rate was 1.0 m L·min-1.The wavelength for ultraviolet detection was 208 nm,the column temperature was 35 ℃,and the injection volume was set at 20 μL.Results：An excellent separation was achieved for 6-sulfocalix（6）arene and its related substances.4-sulfocalix（4）arene was separated well from 6-sulfocalix（6）arene.The assay was validated in the concentration range of 0.1-2 μg（r=0.999 0）.The instrument precision,intra-day precision,inter-day precision and the repeatability were all less than 2%;the content of 6-sulfocalix（6）arene in three batches of crude drugs was in the range of 98.14%-98.65%,while the content of total related substances was not more than 1.0%.Conclusion：The developed method is accurate,reliable,sensitive and specific,which can be used for the determination of 6-sulfocalix（6）arene and the related substances.