how HPLC works - An Overview
how HPLC works - An Overview
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The cellular period carries the sample elements from the column, the place they communicate with the stationary period to varying levels. This conversation decides how long Just about every ingredient spends during the column, resulting in their separation.
각각 다른 산업 분야에 대한 자세한 정보 및 다양한 카테고리는 다음 써모 피셔 사이언티픽 학습 센터에서 산업 및 응용 과학 페이지를 확인하세요.
측정 가능한 농도 범위는 컬럼에 의해서도 결정됩니다. 컬럼 충진제의 종류, 입자 지름, 컬럼의 크기에 따라 분리에 최적인 시료 주입량이 크게 다릅니다.
Right before utilizing a cellular phase solvent we must take away dissolved gases, for example N2 and O2, and compact particulate make any difference, such as dust. Mainly because You will find there's large drop in force across the column—the tension at the column’s entrance is around various hundred atmospheres, however it is atmospheric force on the column’s exit—gases dissolved during the mobile stage are introduced as gasoline bubbles that may interfere Together with the detector’s response.
The 3 pink circles are binary cellular phases produced by combining equivalent volumes of your pure mobile phases. The ternary cellular stage demonstrated via the purple circle has all a few with the pure mobile phases.
The pump is in charge of delivering the cellular section at a continuing circulation rate. This makes certain that the cell section is constantly fed for the column.
규제 약물(마약, 합성 마약, 대마, 각성제, 향정신성 의약품, 아편양제제 등), 반도핑 관련(금지 물질, 금지 약물, 스테로이드 등), 약물 대사물
The elution get of solutes in HPLC is ruled by polarity. For a normal-stage separation, a solute of decreased polarity spends proportionally less time while in the polar stationary stage and elutes in advance of a solute that is certainly a lot more polar. Provided a selected stationary period, retention periods in typical-section HPLC are managed by altering the mobile phase’s Houses. By way of example, If your resolution amongst two solutes is inadequate, switching into a less polar mobile phase retains the solutes to the column for a longer time and offers far more prospect for his or her separation.
The buy of elution of compounds from the column is governed through the depth of connection with more info the stationary stage. The eluent Along with the divided chemicals flows past the detector.
충전제는 실리카겔 혹은 중합체의 미세입자로 표면에 화학 수식이 되어 있는 경우가 대부분이며 여러 종류가 있습니다.
utilizes an autosampler to inject samples. In place of using a syringe to force the read more sample in to the sample loop, the syringe draws sample in the sample loop.
溶媒の組成に勾配を付けて（すなわち組成を連続的に変えて）溶出を行うことも多い。たとえば後述の逆相クロマトグラフィーにおいて水/メタノール勾配を使う場合、まずメタノールの少ない条件で極性の高い物質が溶出し、その後メタノールの割合を増加させてゆくに従ってより極性の低い物質が順次溶出する。これをグラジェント分析と呼ぶ。これに対し、一定組成の溶媒で分析物を溶出させる分析法をアイソクラテック分析と呼ぶ。
Column selection: The stationary period from the column interacts with analytes. Utilizing the Erroneous column chemistry may lead to very poor resolution. Think about using another column with a stationary period that provides improved selectivity for your analytes.
Along with the analysis procedure recognized, let's address popular issues that will come up and the way to troubleshoot them.