Burmaoglu Rifat Emre, Saglik Aslan Serap
Department of Analytical Chemistry, Faculty of Pharmacy, Istanbul University, Beyazit, Istanbul, Turkey.
J Chromatogr Sci. 2019 Jan 1;57(1):33-43. doi: 10.1093/chromsci/bmy078.
A method has been developed and validated for analysis of zoledronic acid (ZOL) and its related substances by ion-pair reversed-phase high performance liquid chromatography with evaporative light scattering detection (ELSD). Chromatographic separation was achieved with gradient elution by using a C18 column, mobile phase containing 12 mM ammonium acetate buffer and 35 mM n-pentylamine, whose pH value is 7.0, and 5% acetonitrile. The mobile-phase flow rate was 1.0 mL/min. The calibration plot was linear in the range from 0.4 mg/mL to 6.0 mg/mL for ZOL and from 6.25 μg/mL to 100 μg/mL for its related substances. ZOL and its related substances, namely imidazole-1-yl-acetic acid, phosphate, phosphite and degradation products did not interfere with each other. The method was rapid, linear, accurate and reproducible. The high performance liquid chromatographic method that has been developed to determine the related substances and assay of ZOL can be used simultaneously to evaluate the quality of regular samples. It can be also used to test the stability samples of ZOL.
已开发并验证了一种采用离子对反相高效液相色谱法结合蒸发光散射检测（ELSD）来分析唑来膦酸（ZOL）及其相关物质的方法。使用C18色谱柱，通过梯度洗脱实现色谱分离，流动相包含12 mM醋酸铵缓冲液和35 mM正戊胺（pH值为7.0）以及5%乙腈。流动相流速为1.0 mL/min。ZOL的校准曲线在0.4 mg/mL至6.0 mg/mL范围内呈线性，其相关物质的校准曲线在6.25 μg/mL至100 μg/mL范围内呈线性。ZOL及其相关物质，即咪唑-1-基-乙酸、磷酸盐、亚磷酸盐和降解产物之间互不干扰。该方法快速、线性、准确且可重现。所开发的用于测定ZOL相关物质和含量的高效液相色谱法可同时用于评估常规样品的质量，也可用于测试ZOL的稳定性样品。
