크로마토그래피 원리의 큰 틀도 마찬가지로 두 상에 대한 분배 차이를 이용하여 분석물을 분리, 정제할 수 있습니다. 다만 크로마토그래피에서 두 개의 상은 하나는 고정하고 다른 하나는 일정 방향으로 이동시켜 사용합니다.
. Solvent triangle for optimizing a reversed-period HPLC separation. The 3 blue circles display cellular phases consisting of an natural solvent and drinking water.
機械的に高い圧力をかけることによって移動相溶媒を高流速でカラムに通し、これにより分析物が固定相に留まる時間を短くして分離能・検出感度を高くすることを特徴とする。
Don't forget, consulting your instrument handbook along with the manufacturer's technical help can even be useful resources when troubleshooting specific concerns with your HPLC system.
). If your detector is usually a diode array spectrometer, then we also can Exhibit The end result as A 3-dimensional chromatogram that demonstrates absorbance being a function of wavelength and elution time.
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Hold a logbook: Doc your observations, such as peak shapes, retention situations, and any improvements designed to the method. This can assist you determine developments and troubleshoot challenges far more effectively.
The pump is the guts with the HPLC system. It provides the mobile section at a continuing and high pressure (approximately 400 atm) from the column. Constant stream fee is critical for acquiring ideal separation and protecting reproducibility. Variables to consider when picking a move price involve:
The info acquisition system controls the HPLC instrument and collects the signal from your detector. This information is displayed being a chromatogram, a graph exhibiting peaks similar to the divided analytes.
An HPLC typically features click here two columns: an analytical column, that's liable for the separation, along with a guard column that is definitely placed before the analytical column to shield it from contamination.
Conversely, a flow amount that may be far too low can cause extreme band broadening. Test your stream fee settings and modify them according to the recognized system.
Common cell section modifiers like acids and bases may be added to fine-tune the conversation between analytes and the column. These modifiers can:
-hydroxybenzoic acid—with a nonpolar C18 column making use of an aqueous buffer click here of acetic acid and sodium acetate as the cellular section. The retention moments for these weak acids are shorter when employing a significantly less acidic cell phase mainly because Each and every solute is existing in an anionic, weak base form that is certainly much less soluble from the nonpolar stationary section.
Whilst Each individual process is exclusive, the following description of the resolve of fluoxetine in serum offers an instructive illustration of a normal treatment. The description here relies on Smyth, W. F. Analytical Chemistry of Sophisticated Matricies