高效液相色谱法测定杯(6)芳烃磺酸钠及其有关物质

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SHISEIDOCAPCELL 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,pH 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 mL·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=0.9990),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, pH=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 mL·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.