ディテクター（検出器）としては目的とする物質の性質に応じて光学的性質（吸光度、屈折率、蛍光等）、電気化学的性質、質量分析法などを利用する装置がある。
ディテクターから出力された、電気信号を記録し、そこからピークを検出、解釈を行う。結果は、感熱紙等に印字される。装置のコントロールをしないのであれば、どのメーカーの物を使用しても問題はないが、通常は、装置のコントロールも同時に行うため、同じメーカーの物を選択する。
측정 가능한 농도 범위는 컬럼에 의해서도 결정됩니다. 컬럼 충진제의 종류, 입자 지름, 컬럼의 크기에 따라 분리에 최적인 시료 주입량이 크게 다릅니다.
Keep in mind, consulting your instrument guide as well as producer's technical help can also be precious means when troubleshooting particular concerns together with your HPLC system.
A reversed-section HPLC separation is completed employing a cellular stage of 60% v/v drinking water and 40% v/v methanol. What is the mobile section’s polarity index?
5.1 displays an illustration of a normal HPLC instrument, which has a number of vital factors: reservoirs that retailer the cell phase; a pump for pushing the cell period from the system; an injector for introducing the sample; a column for separating the sample into its ingredient parts; in addition to a detector for checking the eluent since it comes off the column. Allow’s consider Every single of these factors.
, we could location a solvent proportioning valve prior to one pump. The solvent proportioning value connects two or even more solvent reservoirs on the pump and decides how much of every solvent is pulled through Every single on the pump’s cycles. A different method for eliminating a pulsed move is to incorporate a pulse damper amongst the pump along with the column.
. One issues with an isocratic elution is the fact an acceptable mobile phase toughness for resolving early-eluting solutes might bring about website unacceptably very long retention occasions for late-eluting solutes. Optimizing the cell period for late-eluting solutes, On the flip side, may possibly offer an insufficient separation of early-eluting solutes.
The information acquisition system controls the HPLC instrument and collects the signal in the detector. This information and facts is shown to be a chromatogram, a graph displaying peaks similar to the separated analytes.
(HPLC) we inject the sample, which happens to be in solution sort, into a liquid cell stage. The cell stage carries the sample via a packed or capillary column that separates the sample’s elements based mostly on their own capability to partition involving the cellular period and also the stationary stage. Determine twelve.
If your cell section’s pH is sufficiently acidic, the solutes are present as neutral weak acids that happen to be extra soluble in the stationary stage and consider more time to elute. Because the weak acid solutes do not have identical p
Popular cell period modifiers like acids and bases is often additional to wonderful-tune the interaction in between analytes plus the column. These modifiers can:
The display might be recorded as a series of peaks- each one represents the Each and every ingredient inside the mixture which may take up UV gentle. The area of the peak is proportional to the quantity of the element passed throughout the detector.
Two challenges often shorten the lifetime of the analytical column. Very first, solutes that bind irreversibly to your stationary period degrade the column’s performance by decreasing the amount of stationary phase click here obtainable for effecting a separation. Next, particulate substance injected With all the sample might clog the analytical column.