Boonen Jente, D'hondt Matthias, Veryser Lieselotte, Peremans Kathelijne, Burvenich Christian, Spiegeleer Bart De
Drug Quality and Registration (DruQuaR) Group, Faculty of Pharmaceutical Sciences, Ghent University, Harelbekestraat 72, B-9000 Ghent, Belgium.
Department of Veterinary Medical Imaging and Small Animal Orthopaedics, Faculty of Veterinary Medicine, Ghent University, Salisburylaan 133, B-9820 Merelbeke, Belgium.
J Pharm Anal. 2013 Oct;3(5):330-334. doi: 10.1016/j.jpha.2013.02.002. Epub 2013 Feb 17.
As part of the method development, the injection volume as a critical quality attribute in fast fused-core chromatography was evaluated. Spilanthol, a pharmaceutically interesting -alkylamide currently under investigation in our laboratory, was chosen as the model compound. Spilanthol was dissolved in both PBS and MeOH/HO (70/30, v/v) and subsequently analyzed using a fused-core system hereby selecting five chromatographic characteristics (retention time, area, height, theoretical plates and symmetry factor) as responses. We demonstrated that the injection volume significantly influenced both the qualitative and quantitative performance of fused-core chromatography, a phenomenon which is confounded with the nature of the used sample solvent. From 2 μL up to 100 μL injection volume with PBS as solvent, the symmetry factor decreased favorably by 20%. Moreover, the theoretical plates and the quantitative parameters (area and height) increased up to 30%. On the contrary, in this injection volume range, the theoretical plates for the methanol-based samples decreased by more than 60%, while the symmetry factor increased and the height decreased, both by 30%. The injection volume is thus a critical and often overlooked parameter in fused-core method description and validation.
作为方法开发的一部分，对快速融合核色谱中作为关键质量属性的进样体积进行了评估。我们选择了在实验室中正在研究的一种具有药学意义的烷基酰胺——毛果芸香碱作为模型化合物。将毛果芸香碱溶解在磷酸盐缓冲盐水（PBS）和甲醇/水（70/30，v/v）中，随后使用融合核系统进行分析，在此选择五个色谱特征（保留时间、峰面积、峰高、理论塔板数和对称因子）作为响应指标。我们证明，进样体积显著影响融合核色谱的定性和定量性能，这种现象与所用样品溶剂的性质有关。以PBS为溶剂，进样体积从2 μL增加到100 μL时，对称因子有利地降低了20%。此外，理论塔板数和定量参数（峰面积和峰高）增加了30%。相反，在此进样体积范围内，以甲醇为基质的样品的理论塔板数减少了60%以上，而对称因子增加，峰高降低，两者均降低了30%。因此，进样体积是融合核方法描述和验证中的一个关键且常被忽视的参数。
