反相离子对色谱/蒸发光散射检测器分离唑来膦酸及其有关物质

Separation of Zoledronic Acid and Its Related Substances by Ion-pair Reversed Phase High Performance Liquid Chromatography Coupled with Evaporative Light-scattering Detection

采用反相离子对高效液相色谱/蒸发光散射检测法研究了唑来膦酸及其有关物质的色谱分析与分离方法。优化了色谱条件,固定相为Hypersil C8柱,以含10 mmol/L正戊胺的5 mmol/L乙酸铵缓冲液(用乙酸调节pH至7.0)-甲醇(体积比为97∶3)为流动相,流速为1.0 mL/m in,蒸发光散射检测器检测。在该色谱条件下,唑来膦酸与其有关化合物(包括合成过程中残余的原料咪唑乙酸及分解产物亚磷酸、磷酸盐)的分离良好,唑来膦酸色谱峰与最近杂质峰的分离度大于1.5。本法简便快速,为唑来膦酸的常规分析提供了有效可靠的方法。

A rapid and simple ion-pair reversed phase high performance liquid chromatographic method （ionpair RP-HPLC ） coupled with evaporative light-scattering detection （ELSD） has been established for the routine assay of zoledronic acid and its related substances in bulk material and pharmaceutical dosage forms. Its related substances include remained imidazol-1-yiacetic acid in the synthesis of zoledronic acid and other possible impurities of decomposition such as phosphoric acid and phosphate. The chromatographic conditions were optimized based on satisfactory separation of zoledronic acid from its related substances, reasonable retention time for zoledronic acid and its related substances. The excellent separation of zoledronic acid and its related substances was achieved within 10 min by a Hypersil Cs column with ELSD. The mobile phase was a mixture of methanol （3%） and 5 mmol/L ammonium acetate buffer （97%） containing 10 mmol/L amylamine, and was adjusted to pH 7.0 with acetic acid and delivered at a flow-rate of 1.0 mL/min. The resolution of zoledronic acid peak with next adjacent peak was 〉 1.5.