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首页> 外文期刊>Journal of chromatography, A: Including electrophoresis and other separation methods >The influence of injection volume on efficiency of microbore liquid chromatography columns for gradient and isocratic elution
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The influence of injection volume on efficiency of microbore liquid chromatography columns for gradient and isocratic elution

机译：注射体积对梯度和等异构洗脱微液相色谱柱效率的影响

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摘要

The injection volume and the associated column volume overload is one of the most common issues in miniaturized chromatography. The injection volume should not exceed 10% of the effective column volume. A further reduction of the injection volume leads to an increase in chromatographic efficiency. However, the signal intensity must be above a certain threshold to generate a chromatographic peak that can be detected. Therefore, the injection volume has to be optimized to reach the ideal balance between chromatographic efficiency and sensitivity. This study examined the general influence of the injection volume for both isocratic and gradient elution, depending on the retention factor and peak standard deviation. For this purpose, substances of different polarity were selected to represent a broad elution spectrum. Besides the model analyte naphthalene, these were mainly pharmaceuticals. For all measurements a microbore column with an ID of 300 mu m and packed with 1.9 mu m fully porous particles was used. For isocratic elution, the injection volume was varied between 4 and 16% of the effective column volume. The retention factors were adjusted between 2 and 10. For gradient elution, the injection volume was varied between 4 and 160% of the effective column volume. The observed effects were further investigated using the gradient kinetic plot theory. In isocratic elution, a loss in plate height up to 50% was observed for components that elute near the void time. A significant reduction of the chromatographic efficiency was noticed up to a retention factor of 4. In gradient elution, a reduction in peak capacity could only be observed if the injection volume exceeded 40% of the effective column volume. For some substances, only a slight loss in peak capacity was noticed even with a volume overload of 160%. (C) 2021 Elsevier B.V. All rights reserved.
机译：进样体积和相关的柱体积过载是微型色谱中最常见的问题之一。进样体积不得超过有效柱体积的10%。进样量的进一步减少导致色谱效率的提高。然而，信号强度必须高于某个阈值，才能产生可检测的色谱峰。因此，必须优化进样量，以达到色谱效率和灵敏度之间的理想平衡。本研究考察了注射体积对等度和梯度洗脱的一般影响，这取决于保留因子和峰值标准偏差。为此，选择不同极性的物质来代表广泛的洗脱光谱。除了模型分析物萘，这些主要是药物。所有测量均使用内径为300μm、填充有1.9μm完全多孔颗粒的微孔柱。对于等度洗脱，进样体积在有效柱体积的4%到16%之间变化。保留因子在2到10之间进行调整。对于梯度洗脱，进样体积在有效柱体积的4%到160%之间变化。利用梯度动力学图理论进一步研究了观察到的效应。在等度洗脱中，观察到在空隙时间附近洗脱的组分的板高损失高达50%。当保留因子达到4时，色谱效率显著降低。在梯度洗脱中，只有当进样体积超过有效柱体积的40%时，才能观察到峰容量的降低。对于某些物质，即使在容量过载为160%的情况下，也只注意到峰值容量的轻微损失。（c）2021爱思唯尔B.V.保留所有权利。

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