The Basic Principles Of high performance liquid chromatography
The Basic Principles Of high performance liquid chromatography
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. The working pump as well as equilibrating pump each Have a very piston whose backwards and forwards motion maintains a constant circulation level of up to a number of mL/min and supplies the high output strain necessary to drive the mobile stage with the chromatographic column.
The mobile phase’s stream fee is set with the merged speeds of the two pumps. By changing the relative speeds of The 2 pumps, distinctive binary mobile phases is usually prepared.
, as an example, displays retention moments for 4 weak acids in two cell phases with approximately similar values for (P^ key ). Even though the purchase of elution is the same for both equally mobile phases, each solute’s retention time is afflicted in another way by the choice of natural solvent.
Bubbling an inert gasoline in the mobile phase releases risky dissolved gases. This method is referred to as sparging.
a values, the pH on the mobile stage has another impact on Each individual solute’s retention time, permitting us to find the the best possible pH for effecting an entire separation from the four solutes.
カラム周辺の温度の変動によって溶出時間が安定せず再現性が悪くなる場合があるため、カラム温度を一定に保つために使用する。またカラム温度を分離条件のパラメーターの一つとして積極的に利用する場合もある。
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, a fluorescence detector delivers further selectivity since only a few of the sample’s elements are fluorescent. Detection boundaries are as little as one–ten pg of injected analyte.
The selection of detector relies on the particular requires in the analysis, contemplating elements like sensitivity, selectivity, and compatibility With all the cell stage.
In liquid–liquid chromatography the stationary section can be a liquid movie coated on the packing material, commonly three–ten μm porous silica particles. Because the stationary period could be partly soluble in the cell section, it may well elute, or bleed within the column over time.