The Definitive Guide to how HPLC works
I would really like to enroll in newsletters from Sartorius (Sartorius AG and its affiliated companies) based mostly of my private pursuits.최상의 결과를 위해서는 올바른 시약을 사용함으로써 피크 대칭성을 개선할 수 있습니다.
, as an example, displays retention periods for 4 weak acids in two mobile phases with just about identical values for (P^ prime ). Even though the purchase of elution is identical for both of those cell phases, Each individual solute’s retention time is influenced in a different way by the selection of organic solvent.
, which makes it possible for us to take a look at a wide number of cell phases with only 7 experiments. We commence by adjusting the level of acetonitrile in the cellular stage to provide the best possible separation inside of the desired Evaluation time.
Preserve your instrument: Often cleanse and retain your HPLC system in accordance with the maker's Guidelines. This includes replacing frits, seals, and filters as needed.
분석물의 피크 면적 값(=검출기의 응답)은 정량화를 위해 사용됩니다. 분석자는 분석을 수행하기 전, 분석물의 표준 용액(기지 농도의 시액)을 몇 가지 측정하고, 시료 농도와 획득한 피크 면적 값에 의해 도표된 검량선을 그립니다.
-hydroxybenzoic acid (PH) over a nonpolar C18 column subject to your maximum analysis time website of 6 min. The shaded places stand for regions wherever a separation is not possible, Using the unresolved solutes identified.
The pump is the guts in the HPLC system. It provides the cellular section at a relentless and high strain (as many as four hundred atm) with the column. Reliable movement level is vital for acquiring ideal separation and protecting reproducibility. Things to look at when picking a movement rate involve:
The information acquisition system data and procedures the alerts from your detector, allowing for for your development of chromatograms and also the quantification of compounds.
移動相としては、カラムや装置に悪影響を与えない範囲で各種の溶媒が使用される。水や塩類の水溶液、アルコール類、アセトニトリル、ジクロロメタン、トリフルオロ酢酸などが用いられる。相溶性のある（互いに混じり合う）溶媒を混合して使用する場合が多い。
The cellular section’s move amount is set by the blended speeds of the two pumps. By altering the relative speeds of The 2 pumps, various binary mobile phases may be prepared.
高速液体クロマトグラフィー 高速液体クロマトグラフィー（こうそくえきたいクロマトグラフィー、英: high performance liquid chromatography、略称: HPLC）はカラムクロマトグラフィーの一種である。移動相として高圧に加圧した液体を用いることが特徴である。
The elution get of solutes in HPLC is ruled by polarity. For a normal-stage separation, a solute of decreased polarity spends proportionally fewer time during the polar stationary period and elutes prior to a solute that may be additional polar. Presented a specific stationary phase, retention situations in standard-phase HPLC are managed by modifying the cell website stage’s Attributes. For example, Should the resolution in between two solutes is poor, switching to your much less polar cellular section keeps the solutes about the column for an extended time and gives extra possibility for his or her separation.
The liquid that transports the sample from the column is called the cell period. It comprises of a number of solvents decided on dependant on the analysis’s exclusive demands.