Vandercruyssen Kirsten, D'Hondt Matthias, Vergote Valentijn, Jansen Herwig, Burvenich Christian, De Spiegeleer Bart
Drug Quality and Registration (DruQuaR) group, Faculty of Pharmaceutical Sciences, Ghent University, Harelbekestraat 72, B-9000 Ghent, Belgium.
Dafra Pharma International, Slachthuisstraat 30/7, B-2300 Turnhout, Belgium.
J Pharm Anal. 2014 Feb;4(1):37-52. doi: 10.1016/j.jpha.2013.03.006. Epub 2013 Apr 25.
A highly selective and stability-indicating HPLC-method, combined with appropriate sample preparation steps, is developed for β-artemether assay and profiling of related impurities, including possible degradants, in a complex powder for oral suspension. Following HPLC conditions allowed the required selectivity: a Prevail organic acid (OA) column (250 mm×4.6 mm, 5 μm), flow rate set at 1.5 mL/min combined with a linear gradient (where =25 mM phosphate buffer (pH 2.5), and =acetonitrile) from 30% to 75% in a runtime of 60 min. Quantitative UV-detection was performed at 210 nm. Acetonitrile was applied as extraction solvent for sample preparation. Using acetonitrile-water mixtures as extraction solvent, a compartmental behaviour by a non-solving excipient-bound fraction and an artemether-solubilising free fraction of solvent was demonstrated, making a mobile phase based extraction not a good choice. Method validation showed that the developed HPLC-method is considered to be suitable for its intended regulatory stability-quality characterisation of β-artemether paediatric formulations. Furthermore, LC-MS on references as well as on stability samples was performed allowing identity confirmation of the β-artemether related impurities. MS-fragmentation scheme of β-artemether and its related substances is proposed, explaining the / values of the in-source fragments obtained.
开发了一种高选择性和稳定性指示的高效液相色谱法，并结合适当的样品制备步骤，用于测定复方口服混悬液中β-蒿甲醚的含量以及分析包括可能的降解产物在内的相关杂质。以下高效液相色谱条件可实现所需的选择性：Prevail有机酸（OA）柱（250 mm×4.6 mm，5 μm），流速设定为1.5 mL/min，结合线性梯度（其中 = 25 mM磷酸盐缓冲液（pH 2.5）， = 乙腈），在60分钟的运行时间内从30%到75%。在210 nm处进行定量紫外检测。乙腈用作样品制备的萃取溶剂。使用乙腈-水混合物作为萃取溶剂时，证明了非溶解辅料结合部分和蒿甲醚增溶游离部分溶剂存在分室行为，这使得基于流动相的萃取不是一个好选择。方法验证表明，所开发的高效液相色谱法被认为适用于其预期的β-蒿甲醚儿科制剂的监管稳定性质量表征。此外，对对照品以及稳定性样品进行了液相色谱-质谱联用分析，以确认β-蒿甲醚相关杂质的身份。提出了β-蒿甲醚及其相关物质的质谱裂解方案，解释了所获得的源内碎片的 / 值。
