Jimidar M Ilias, De Smet Maurits, Sneyers Rudy, Van Ael Willy, Janssens Willy, Redlich Dirk, Cockaerts Paul
Johnson & Johnson Pharmaceutical Research and Development, A Division of Janssen Pharmaceutica N.V., Global Analytical Development-Method Development, Beerse, Belgium.
J Capill Electrophor Microchip Technol. 2003 May-Aug;8(3-4):45-52.
High-performance liquid chromatography is usually used to assay the main compound and organic impurity content of drug substance and drug product during pharmaceutical development. A crucial validation parameter of these methods is specificity--the ability to unequivocally assess the analyte in the presence of component expected to be present. Typically, these include impurities, degradation products, and matrices. Besides adequate chromatographic separation with sufficient selectivity, additional 2- or 3-D spectroscopic or chromatographic tools are frequently necessary for this purpose. In our current practice, HPLC is used with ultraviolet photodiode array detection and on-line mass spectrometry (LC-UVDAD-MS) during the assessment of specificity. Although this approach is very powerful and can solve the majority of problems, separation of isomers of the main compound is still difficult. Since HPLC usually cannot offer the required selectivity and because of the similar molecular weights, structural isomers are not specifically detected using LC-MS. Capillary electrophoresis, on the other hand, offers high separation efficiency and can be applied as an adjunct to HPLC. Therefore, a set of highly selective CE methods is used orthogonally in the specificity assessment of HPLC methods.
在药物研发过程中，高效液相色谱法通常用于测定原料药和制剂中主要化合物及有机杂质的含量。这些方法的一个关键验证参数是专属性，即在预期存在的组分存在下明确评估分析物的能力。通常，这些组分包括杂质、降解产物和基质。除了具有足够选择性的充分色谱分离外，为此目的通常还需要额外的二维或三维光谱或色谱工具。在我们目前的实践中，在评估专属性时，高效液相色谱法与紫外光电二极管阵列检测和在线质谱联用（LC-UVDAD-MS）。虽然这种方法非常强大，可以解决大多数问题，但主要化合物异构体的分离仍然很困难。由于高效液相色谱法通常无法提供所需的选择性，并且由于分子量相似，结构异构体无法使用液相色谱-质谱法进行特异性检测。另一方面，毛细管电泳具有高分离效率，可作为高效液相色谱法的辅助方法。因此，一组高选择性的毛细管电泳方法被正交用于高效液相色谱法的专属性评估。
