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。
In this particular instrument, Just about every pump sends its mobile section to a mixing chamber in which they Blend to form the final cellular stage. The relative pace of the two pumps determines the mobile period’s remaining composition.
The area of the height is mechanically detected by the computer. The computer also detect the retention time of that precise element.
The analysis is complicated via the advanced matrix of serum samples. A solid-period extraction followed by an HPLC Investigation utilizing a fluorescence detector offers the required selectivity and detection restrictions.
A reversed-period HPLC separation is carried out using a mobile phase of sixty% v/v drinking water and 40% v/v methanol. What's the mobile period’s polarity index?
A detector identifies and steps Every single part. Retention time indicates some time taken for each compound to exit the column. HPLC's efficiency is determined by things like column sort and cell phase composition. Regular upkeep makes certain correct final results. Comprehension HPLC's phase-by-phase process is important for specific chemical Investigation in laboratories.
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前述した従来の順相タイプに対して、逆相クロマトグラフィーにおいては固定相に低極性のもの（例えばシリカゲルにアルキル基を共有結合させたもの）を、移動相に高極性のもの（例えば水や塩類の水溶液、アルコール、アセトニトリルなどの有機溶媒）を用いる。また珍しいケースではあるが、分離のための移動相pHをシリカゲルの使用範囲から外れたところに設定する必要がある場合、あるいはシリカゲル表面に残っている未反応シラノール基が分離に悪影響を及ぼし、かつそれが移動相の変更によっても解決できない場合には、固定相として樹脂を用いることがある。分析物はより極性の低いほどより強く固定相と相互作用して溶出が遅くなる。また極性の低い物質の割合が多い移動相ほど溶出が早くなる。
The information acquisition system controls the HPLC instrument and collects the signal with the detector. This information and facts is shown as a chromatogram, a graph check here exhibiting peaks corresponding to the divided analytes.
Retention periods: The time it's going to take for every analyte to get to the detector, providing a attribute fingerprint for identification.
The HPLC column properties the stationary stage, a crucial component for separating analytes. Selecting the right column is crucial:
Compounds within the sample partition concerning the stationary period and the cellular phase in partition chromatography. Compounds that has a more powerful affinity for your stationary phase commit much more time interacting with it, leading to slower elution from the column.
-hydroxybenzoic acid—on the nonpolar C18 column using an aqueous buffer of acetic acid and sodium acetate as being the mobile stage. The retention situations for these weak acids are shorter when utilizing a fewer acidic mobile period because Just about every solute is present in an anionic, weak foundation form that is definitely much less soluble from the nonpolar stationary phase.
The get more info selection to get started with acetonitrile is arbitrary—we are able to just as simply pick out to start with methanol or with tetrahydrofuran.