物質の持つ特定波長の光を吸収する性質を利用した検出器。次のようなものが存在している。
The sample injector is used to inject the sample into the HPLC system. To realize ideal elution, the sample is normally dissolved in an appropriate solvent that matches the mobile stage.
. HPLC separation of a mix of flavonoids with UV/Vis detection at 360 nm and, within the inset, at 260 nm. The choice of wavelength impacts Each individual analyte’s signal.
Non-polar molecules are slowed down on their own way from the column. They type various levels of attraction With all the hydrocarbon teams principally via van der Waals dispersion forces and hydrophobic interactions.
Degassing is accomplished in various techniques, but the most common are using a vacuum pump or sparging having an inert gasoline, for instance He, which has a low solubility within the cellular stage. Particulate products, which can clog the HPLC tubing or column, are taken out by filtering the solvents.
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The interface among the HPLC along with the mass spectrometer is technically tougher than that inside of a GC–MS as a result of incompatibility of the liquid cell period Along with the mass spectrometer’s high vacuum requirement.
It achieves this by exploiting the click here differing interactions of sample compounds with two important phases: the mobile phase as well as stationary phase. Knowledge the Main factors of the HPLC system as well as their roles is important for prosperous Investigation.
The determine below displays the calibration curve and calibration equation for that list of external standards. Substituting the sample’s peak space into the calibration equation offers the focus of caffeine inside the sample as 94.four mg/L.
This leads to various elution fees for the several parts and results in the separation in the components since they flow out the column. In comparison with column chromatography, HPLC is highly automated and extremely delicate.
, that's the more prevalent sort of HPLC, here the stationary phase is nonpolar as well as the mobile section is polar. The most typical nonpolar stationary phases use an organochlorosilane where by the R group is definitely an n
The region below Every single peak is proportional to the quantity of the corresponding analyte. The information acquisition system allows for the Evaluation of peak retention occasions, peak locations, and also the calculation of analyte concentrations.
The selection of detector depends on the specific requirements from the Evaluation, thinking about aspects like sensitivity, selectivity, and compatibility With all the mobile period.
In liquid–liquid chromatography the stationary stage is often a liquid film coated on the packing materials, generally 3–10 μm porous silica particles. Since the stationary phase might be partly soluble inside the mobile stage, it might elute, or bleed with the column with time.