Timothy Cross Liquid chromatography is just liquid chromatography, right? Conversely with analytical LC the goal is to identify and perform qualitative and quantitative analysis of the compound s in the sample mixture. Google is a trademark of Google LLC. Google on Google LLC kaubamärk. Analytical LC is often used to assess the purity and purification performed by preparative LC and preparative applications are often initially performed at an analytical level and then scaled-up to the preparative scale.
Columns and Chemistries In preparative LCwhere the primary goal is to separate and collect pure protein with a high yield as fast as possible then this dictates a larger column diameter, typically in the range of 50 — mm to achieve the required throughput and capacity. To increase the capacity of the column the options are to increase the diameter or the length of the column increasing column length can also aid purity.
The stationary phase packed in to the column are typically 10 microns and above and permit flow rates in the tens of millilitres per minute depending on the column size and throughput required. When all of these parameters are combined together the observed chromatogram tends to contain peaks that are broader and less defined than in analytical LC. As you would expect, analytical LC is almost the exact opposite as the goal is centred on clearly resolving individual peaks and having a high peak capacity for identification and analysis.
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Typical analytical LC columns have a diameter between 4. With smaller inner diameter columns and smaller particles comes a reduction in flow rate, typically under 2 millilitres per minute, and sharper, more defined, peaks in the chromatogram.
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The smaller column diameters and particles, in conjunction with a relatively high flow rate, in proportion give a much higher backpressure which characterises analytical LC and why it is often also referred to as HPLC. The goals of the two approaches do favour particular chemistries and applications though. In preparative LC, where purity and yield are important, then affinity resins such as Protein A are common, especially in protein purification where highly specific binding can aid purity.
In analytical LC, reverse-phase chemistries such as C18 are the first-choice with mixed-mode chemistries gaining popularity for demanding separations, however all chemistries can be run on both, after method development and optimisation.
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Instrument Differences The biggest difference between a preparative and analytical LC system is the system backpressures. In analytical LC, with relatively high flow rates compared to small column diameters and particles, backpressures can get very high.
Conversely on standard preparative LC systems that utilise large column diameters and particles, 10 bar and under is more common.
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Dispersion is a common problem for both techniques, but is more of an issue with analytical LC where sharp, well-resolved peaks from complex mixtures are required compared to affinity-based preparative separation where some peak broadening is less of a problem. This considerably increases the backpressure and is why wider tubing is used in preparative LC. Additional modules also differ in preparative and analytical LC systems.
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