Effect of column dimension on observed column efficiency in very high pressure liquid chromatography
Effect of column dimension on observed column efficiency in very high pressure liquid chromatography
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DOI：
10.1016/j.chroma.2012.05.054
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发表时间：
2012-10-26
影响因子：
4.1
通讯作者：
Bradley, Ashley C.
中科院分区：
文献类型：
--
作者：
Wu, Naijun;Bradley, Ashley C.
研究了超高压液相色谱中不同内径色谱柱的柱外体积对观察到的线速度的影响。结果表明，观测到的线速度约为4.5、9.5、16.8。分别比 4.6、3.0、2.1 和 1.0 mm 内径色谱柱的柱外体积贡献校正线速度低 39.5%。使用不同内径的 50 cm 长管道获得了柱外谱带展宽和柱外体积之间的经验关系。对于内径为 0.0635-0.178 毫米（0.025-0.07 英寸）的管道，柱外体积的峰值方差与柱外体积的平方几乎成线性比例。使用 50/50 乙腈/水流动相，流速大于 0.3 mL/min。使用具有四种不同柱内径的 50 mm 柱和具有三种不同长度的 2.1 mm 内径柱，研究了柱内径和柱长度对观察效率的影响。结果表明，对于二苯甲酮 (k = 5.5)，3.0、2.1 和 1.0 mm 内径色谱柱的观察到的柱效分别比 4.6 mm 内径色谱柱低 18%、33% 和 73%。与填充 5.0 μm 颗粒的 150 mm x 2.1 mm 色谱柱相比，使用填充 1.7 μm 颗粒的 50 mm x 2.1 mm 色谱柱观察到苯丙酮 (k = 3.3) 的理论塔板数减少了约 20%，而前一色谱柱的分离速度提高了 9 倍。决定 VHPLC 中柱外谱带展宽程度的是柱与柱外体积之比，而不是绝对柱外体积。 (C) 2012 Elsevier B.V. 保留所有权利。
The effect of extra-column volume on observed linear velocity was investigated for columns of various internal diameters in very high pressure liquid chromatography. The results showed that the observed linear velocities were approximately 4.5, 9.5, 16.8. and 39.5% lower than the linear velocities corrected for the extra-column volume contribution for 4.6,3.0,2.1, and 1.0 mm internal diameter columns, respectively. An empirical relationship between extra-column band broadening and extra-column volume was obtained using 50 cm long tubings of various internal diameters. The peak variance from the extra-column volume is near linearly proportional to the square of the extra-column volume for tubings with 0.0635-0.178 mm (0.025-0.07 in.) i.d. using a 50/50 acetonitrile/water mobile phase at flow rates greater than 0.3 mL/min. The effect of column internal diameter and column length on observed efficiency was studied using 50 mm columns with four different column internal diameters and 2.1 mm i.d columns with three different lengths. The results showed that the observed column efficiencies for 3.0, 2.1, and 1.0 mm internal diameter columns were 18, 33, and 73% lower than that for a 4.6 mm internal diameter column for benzophenone (k = 5.5), respectively. An approximate 20% decrease in theoretical plate number was observed for propiophenone (k = 3.3) using a 50 mm x 2.1 mm column packed with 1.7 mu m particles compared to a 150 mm x 2.1 mm column packed with 5.0 mu m particles, while the former column provided 9 fold faster separation. It is the column to extra column volume ratio instead of absolute extra-column volume that determines the degree of extra-column band-broadening in VHPLC. (C) 2012 Elsevier B.V. All rights reserved.