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The column dimension is the same. The column is stuffed with silica particles that are modified to make them non-polar. This is finished by attaching very long hydrocarbon chains (eight–eighteen C atoms) to its surface.
Being a common rule, a two unit modify during the polarity index corresponds to an roughly ten-fold transform inside of a solute’s retention aspect. Right here is a straightforward illustration. If a solute’s retention factor, k
Shifting the mobile period’s composition given that the separation progresses is just one Resolution to this issue. To get a reversed-stage separation we use an First cellular section that may be far more polar. As being the separation progresses, we regulate the composition of cellular period so that it gets to be considerably less polar (see Figure 12.five.6
Inside the column, separation happens determined by the differential interactions between analytes as well as the stationary stage. Analytes website by using a stronger affinity for the stationary phase move slower from the column in comparison to Individuals with weaker interactions.
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
The functioning pressure inside an HPLC is sufficiently high that we can not inject the sample to the cellular stage by inserting a syringe by way of a septum, as is achievable in gasoline chromatography. As an alternative, we inject the sample utilizing a loop injector
This big difference in interaction situations brings about the separation of analytes because they exit the column at distinct occasions.
移動相としては、カラムや装置に悪影響を与えない範囲で各種の溶媒が使用される。水や塩類の水溶液、アルコール類、アセトニトリル、ジクロロメタン、トリフルオロ酢酸などが用いられる。相溶性のある（互いに混じり合う）溶媒を混合して使用する場合が多い。
Should the cell section’s pH is adequately acidic, the solutes are existing as neutral weak acids which are a lot more soluble while in the stationary section and choose longer to elute. Because the weak here acid solutes don't have identical p
When the cellular section’s pH is adequately acidic, the solutes are present as neutral weak acids that happen to be a lot more soluble during the stationary stage and choose for a longer time to elute. As the weak acid solutes would not have identical p
A reversed-stage HPLC separation is performed utilizing a cell stage of sixty% v/v water and forty% v/v methanol. What is the cellular phase’s polarity index?
Decreasing the amount of acetonitrile and growing the level of water within the mobile will maximize retention periods, delivering much more time to influence a separation.