División Química Analítica y CIDEPINT, Facultad de Ciencias Exactas, Universidad Nacional de La Plata, La Plata, Argentina.
J Sep Sci. 2010 Sep;33(17-18):2645-53. doi: 10.1002/jssc.201000254.
Recent developments in HPLC methods have focused on various strategies in order to increase the speed of analysis. One area of impressive growing is column technology. Today, analytical methods that propose the use of short columns packed with sub-2 μm particles installed in ultra high-pressure LC instruments are not uncommon. Another strategy consisted of heating thermally resistant columns to temperatures well above of 100°C in order to reduce eluent viscosities and, therefore, column backpressure. We discuss experimental conditions for achieving high-throughput analysis using standard instruments with a few simple modifications. The chromatographic performance of two particulated and a silica-based monolithic column operated at moderate temperatures and flow rates are compared. The monolithic column proved to be stable over several thousands column volumes at 60°C. More important, its resistance to mass transfer at this temperature was significantly reduced. Very fast separations of two different mixtures of pharmaceutical compounds, anti-inflammatory drugs and β-blockers, were achieved with the three columns at 60°C by using ACN/buffer at 5 mL/min. Excellent peak shapes of basic solutes and quite reasonable resolutions were achieved in very short analysis times with columns operated at temperatures moderately higher than the usual room temperature.
近年来，高效液相色谱方法的发展集中在各种策略上，以提高分析速度。一个令人印象深刻的发展领域是柱技术。如今，分析方法建议使用短柱，柱内填充亚 2μm 颗粒，并安装在超高压液相色谱仪器中，这种方法已很常见。另一种策略是将热稳定柱加热到 100°C 以上的温度，以降低洗脱液的粘度，从而降低柱背压。我们讨论了使用标准仪器通过一些简单的修改来实现高通量分析的实验条件。比较了在中等温度和流速下操作的两种颗粒状和基于硅胶的整体式柱的色谱性能。整体式柱在 60°C 下经过数千个柱体积后仍保持稳定。更重要的是，其在该温度下的传质阻力显著降低。通过使用 ACN/缓冲液在 5 mL/min 的流速下，在 60°C 下，三种柱可实现两种不同药物混合物、抗炎药和β受体阻滞剂的快速分离。在稍高于室温的温度下运行的柱可在非常短的分析时间内实现基本溶质的出色峰形和相当合理的分辨率。
