The Basic Principles Of high performance liquid chromatography
The Basic Principles Of high performance liquid chromatography
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HPLC works following The essential theory of thin layer chromatography or column chromatography, where it has a stationary section in addition to a cell stage. The cell section flows from the stationary period and carries the elements with the combination with it.
최상의 결과를 위해서는 올바른 시약을 사용함으로써 피크 대칭성을 개선할 수 있습니다.
예를 들어 설탕과 같이 물에 녹기 쉬운 물질을 첨가했을 때 설탕은 기름층에 거의 녹지 않으므로 물층에 많이 존재하게 됩니다. 반대로 식용유와 같이 헥산에 녹기 쉬운 용질을 첨가했을 때는 물층보다 기름층에 많이 존재합니다. 이와같이, 설탕과 식용유는 물과 헥산의 두 상 사이의 존재의 비율(=분배 비율)이 크게 다르기 때문에, 만약 당신과 이 분액깔대기에서 설탕만을 분리하고 싶다면, 분액깔대기에서 물층만을 꺼내 물을 증류시키면 설탕만을 얻을 수 있습니다.
By adhering to these guidelines and systematically addressing potential results in, you could properly troubleshoot frequent HPLC complications and be certain your analyses are correct and reputable.
. Illustration of an average high-performance liquid chromatograph with insets showing the pumps that move the cell phase in the system as well as the plumbing utilized to inject the sample in to the cell section.
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The column is full of a stationary read more stage content. The selection of column and stationary period is dependent upon the character in the compounds currently being analyzed and also the separation ambitions.
高速液体クロマトグラフィーにおいては各物質は比較的鋭いピークとして検出され、分離（他の物質のピークと明確に分けられる）および検出（鋭いピークにより高い感度が得られる）の能力が従来の液体クロマトグラフィーより良くなる。
-hydroxybenzoic acid—with a nonpolar C18 column employing an aqueous buffer of acetic acid and sodium acetate since the cell stage. The retention periods for these weak acids are shorter when employing a significantly less acidic mobile stage simply because Every single solute is current in an anionic, weak foundation form that is certainly a lot less soluble during the nonpolar stationary period.
). Because the tubing and fittings that carry the cell section have pressure boundaries, a higher back again stress demands a reduce circulation price and a longer Assessment time. Monolithic columns, during which the good support is just one, porous rod, give column efficiencies akin to a packed capillary column though permitting for a lot quicker circulation costs. A monolithic column—which generally is comparable in sizing to a standard packed column, Even though scaled-down, capillary columns also are offered—is prepared by forming the mono- lithic rod inside a mildew and covering it with PTFE tubing or possibly a polymer resin.
The mobile phase’s flow level is set by the combined speeds of The 2 pumps. By shifting the relative speeds of The 2 pumps, diverse binary cell phases is usually prepared.
Quite a few differing kinds of detectors are working of hplc system use to monitor HPLC separations, the vast majority of which utilize the spectroscopic approaches from Chapter 10 or perhaps the electrochemical approaches from Chapter eleven.
, for example, has two cellular section reservoirs which are used for an isocratic elution or possibly a gradient elution by drawing solvents from 1 or each reservoirs.
A quantitative HPLC analysis is often easier than the usual quantitative GC Assessment due to the fact a fixed quantity sample loop provides a more specific and precise injection.