HPLC法测定甲氧沙林搽剂的有关物质

Determination of Related Substances in Methoxsalen Solution for Topical by HPLC

目的采用高效液相色谱(HPLC)法测定甲氧沙林搽剂的有关物质。方法用十八烷基硅烷键合硅胶为填充剂(Waters Nova-Pak C18,150 mm×3.9 mm,4μm),以水为流动相A,乙腈为流动相B,梯度洗脱,检测波长254 nm。以外标法测定补骨脂素和佛手柑内酯,限度均为1.0%;以主成分自身对照法测定其他未知杂质,单个未知限度为0.2%,未知杂质峰面积的和限度为1.0%。结果甲氧沙林、补骨脂素、佛手柑内酯之间的分离度及线性关系良好,相关系数均为1.000;甲氧沙林、补骨脂素、佛手柑内酯的检出限分别为0.023μg·m L^(-1)、0.043μg·m L^(-1)及0.040μg·m L^(-1);定量限分别为0.076μg·m L^(-1)、0.141μg·m L^(-1)及0.132μg·m L^(-1),供试品溶液在24 h内稳定,且辅料无干扰。结论所建立方法的灵敏度和准确度均达到有关物质检查的要求,适用于甲氧沙林搽剂有关物质的检测。

OBJECTIVE To use HPLC method to determine the related substances in Methoxsalen Solution For Topical.METHODS HPLC was adopted,the determination was performed on C18（ Waters Nova-Pak C18,150 mm×3.9 mm,4 μm）,water was the mobile phase A,acetonitrile was the mobile phase B,determined the related substances by gradient elution,ultraviolet detection wavelength was 254 nm.Determination of psoralen and bergapten by external standard method,the limit was 1.0%; determination of other unknown impurities by principal component self-control method,the single unknown limit was 0. 2%,the sum of the unknown impurity peak area was 1.0%. RESULTS The resolution and linearity between methoxsalen,psoralen and bergapten were good,the correlation coefficient were r = 1.000; the detection limit of methoxsalen,psoralen and bergapten were 0.023 μg·m L^（-1）,0.043μg·m L^（-1）,0.040 μg·m L^（-1）;the limit of quantitation were 0.076μg·m L^（-1）,0.141 μg·m L^（-1）,0.132μg·m L^（-1）,the test solution were stable in 24 hours,and the auxiliary materials were noninterference. CONCLUSION The sensitivity and accuracy of the method have reached the requirements of the related material inspection,and are suitable for the determination of related substances in Methoxsalen Solution For Topica.