Thermodynamic interpretation of the drift and noise of gradient baselines in reversed-phase liquid chromatography using mobile phase additives

化学 色谱法 口译(哲学) 相(物质) 高效液相色谱法 反相色谱法 噪音(视频) 有机化学 人工智能 计算机科学 图像(数学) 程序设计语言
作者
Fabrice Gritti
出处
期刊:Journal of Chromatography A [Elsevier BV]
卷期号:1633: 461605-461605 被引量:6
标识
DOI:10.1016/j.chroma.2020.461605
摘要

Abstract The drift and noise of acetonitrile-water gradient baselines (5-95%, v/v, 5 min linear gradient) in reversed phase liquid chromatography (RPLC) are recorded at a wavelength of 215 nm using 0.1% trifluoroacetic acid (TFA) as the mobile phase additive, a 4.6 mm  ×  150 mm 5 μm Symmetry-C18 RPLC column, and an Arc system (low-pressure gradient proportioning valve or GPV, pump with a stroke volume of either 66 or 132 μL, no mixer) as the LC instrument. These observations are predicted from solid-liquid adsorption thermodynamics which requires the measurement of the excess adsorption isotherm of acetonitrile from water onto the RPLC column and of the variation of the Henry’s constant of TFA as a function of the volume fraction of acetonitrile in the bulk mobile phase. The incomplete mixing of the acetonitrile and water packets delivered by the low-pressure GPV is represented by a sinusoidal perturbation of the programmed volume fraction of acetonitrile during the entire gradient. The variation of the TFA absorbance at 215 nm with increasing acetonitrile concentration is measured in order to transform TFA concentration into the observable absorbance unit. The drift and noise of the gradient baseline are calculated by solving numerically (Rouchon method) the equilibrium-dispersive (ED) mass balance equations of acetonitrile and TFA. The agreement between the calculated and observed gradient baselines is very good as the proposed model of chromatography accurately accounts for the displacement of TFA between stationary and mobile phases (early excess and late deficit of TFA concentration relative to 0.1%) and for the frequency (equal to the ratio of the applied flow rate to the stroke volume) and the amplitude of the periodic noise recorded during the gradient. From a practical viewpoint, the drift of the gradient baseline can be minimized by maximizing the ratio of the gradient volume to the hold-up volume ( > 10) and/or by minimizing the retention factor of the mobile phase additive in the water-rich eluent (k
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