The Basic Principles Of high performance liquid chromatography
The Basic Principles Of high performance liquid chromatography
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To prevent the loss of stationary stage, which shortens the column’s lifetime, it truly is sure covalently on the silica particles. Bonded stationary phases
A number of other detectors are already Employed in HPLC. Measuring a improve while in the cell section’s refractive index is analogous to checking the cellular phase’s thermal conductivity in gas chromatography. A refractive index detector is almost common, responding to Virtually all compounds, but has a relatively lousy detection limit of 0.
a values, the pH on the mobile stage has a unique effect on Just about every solute’s retention time, making it possible for us to locate the ideal pH for effecting a complete separation of your four solutes.
The selection to begin with acetonitrile is arbitrary—we can just as effortlessly choose to begin with methanol or with tetrahydrofuran.
A reversed-stage HPLC separation is carried out using a mobile period of 60% v/v h2o and forty% v/v methanol. Exactly what is the cell stage’s polarity index?
シリカゲルの粒子径が小さければ小さいほどピークの分離性は良くなるが、送液に必要なポンプの圧力が高くなる。そのため、ポンプ－インジェクター間、インジェクター－カラム間の配管の耐圧を上げたり、カラム自体を比較的高温の下にさらして溶媒の粘度を下げ、抵抗を小さくする工夫をしている。
규제 약물(마약, 합성 마약, 대마, 각성제, 향정신성 의약품, 아편양제제 등), 반도핑 관련(금지 물질, 금지 약물, 스테로이드 등), 약물 대사물
-hydroxybenzoic acid elutes additional little read more by little. Although we will solve absolutely these two solutes working with cell stage that is certainly sixteen% v/v acetonitrile, we can not solve them When the mobile phase is ten% tetrahydrofuran.
Bad resolution usually means analytes elute as well shut together, earning them difficult to distinguish. Here is the way to read more troubleshoot:
원하는 분석 결과를 얻기 위해서는 컬럼도 충분히 고려하고 선택하는 것이 좋습니다.
The HPLC column properties the stationary stage, a critical ingredient for separating analytes. Choosing the appropriate column is vital:
During this segment we look at the essential plumbing needed to transfer the mobile phase with the column and to inject the sample to the cell section.
The detector screens the eluent as it exits the column. Various detectors are made use of depending on the compounds currently being analyzed and the expected sensitivity.
, for example, displays an amperometric flow mobile. Effluent with the column passes about the working electrode—held at a constant possible relative to your downstream reference electrode—that absolutely oxidizes or decreases the analytes.