Department of Chemistry, BITS-Pilani Hyderabad Campus, Jawaharnagar, Hyderabad, Medchal (Dist), Telangana 500078, India.
Department of Pharmacy, BITS-Pilani Hyderabad Campus, Jawaharnagar, Hyderabad, Medchal (Dist), Telangana 500078, India.
Ann Pharm Fr. 2022 Jul;80(4):448-459. doi: 10.1016/j.pharma.2021.11.005. Epub 2021 Dec 9.
The objective of the current study was to develop and validate the sensitive LC-MS methods for trace analysis of genotoxic impurities in Ivacaftor and Lumacaftor. The first method is for the trace analysis of 2,4-di-tert-butyl-5-nitrophenol in ivacaftor and the second method is for the trace analysis of 1-(2,2-difluoro-1,3-benzodioxol-5yl)-cyclopropane carboxylic acid and 3-carboxyphenyl boronic acid in lumacaftor.
High pure analytical grade solvents and reagents were used for this study. The chromatographic separation was performed on Luna C18 (250×4.6mm, 5.0μm) at a column temperature of 25°C using eluent consisting of acetonitrile and 0.1% v/v formic acid in water in a gradient elution mode. The eluent was run at a flow of 1.0mL/min and injection volume of 20μL.
The linearity, precision and accuracy of the developed methods was validated over the concentration range of 0.35-15.0ppm for 2,4-di-tert-butyl-5-nitrophenol, 0.30-15.0ppm for 1-(2,2-difluoro-1,3-benzodioxol-5yl)-cyclopropane carboxylic acid and 0.23-15.0ppm for 3-carboxyphenyl boronic acid. In both methods, interference was not observed at the retention time of analyte peaks. All the analytes were found to be stable in solution for a period of 48h.
The proposed methods are reliable, sensitive, precise, accurate, and robust for the trace level quantification of genotoxic impurities in Ivacaftor and Lumacaftor. These methods can be successfully implemented in the quality control lab for routine analysis.
本研究旨在开发和验证灵敏的 LC-MS 方法，用于痕量分析伊伐卡托和卢美卡托中的遗传毒性杂质。第一种方法用于痕量分析伊伐卡托中的 2,4-二叔丁基-5-硝基苯酚，第二种方法用于痕量分析卢美卡托中的 1-(2,2-二氟-1,3-苯并二氧戊环-5-基)-环丙烷羧酸和 3-羧基苯硼酸。
本研究使用高纯分析级溶剂和试剂。色谱分离在 Luna C18（250×4.6mm，5.0μm）柱上进行，柱温为 25°C，洗脱剂为乙腈和 0.1%v/v 甲酸水溶液，采用梯度洗脱模式。洗脱液流速为 1.0mL/min，进样量为 20μL。
在 2,4-二叔丁基-5-硝基苯酚的浓度范围为 0.35-15.0ppm、1-(2,2-二氟-1,3-苯并二氧戊环-5-基)-环丙烷羧酸的浓度范围为 0.30-15.0ppm、3-羧基苯硼酸的浓度范围为 0.23-15.0ppm 时，所建立的方法具有良好的线性、精密度和准确度。在两种方法中，在分析物峰的保留时间处均未观察到干扰。所有分析物在溶液中 48h 内均稳定。
所提出的方法可靠、灵敏、精确、准确、稳健，可用于痕量定量分析伊伐卡托和卢美卡托中的遗传毒性杂质。这些方法可成功应用于质量控制实验室进行常规分析。
