HPLC溶剂选择中的批判考虑因素

Solvent selection for mobile phase preparation requires careful review of its properties
Solvent selection for mobile phase preparation requires careful review of its properties
Solvent selection for mobile phase preparation requires careful review of its properties

选择溶剂用作流动阶段HPLC.分析是方法开发的关键组成部分。不可能根据分析要求确定将满足所有应用的通用溶剂,并且通常是溶剂的组合。选择合适的溶剂是基于它们的物理性质和叠加能力,并柱固定相。

本文讨论了一些重要的考虑因素,在决定溶剂作为流动阶段的适用性方面发挥着重要作用。

Cost

成本是,随着HPLC需要优异的纯度等级溶剂,且很常见于大型实验室中的时钟运行的数十种HPLC系统。这意味着在批量数量中消耗高级溶剂,因此成本考虑起到重要作用。

溶解度

样品应完全溶于流动阶段。最轻微的不透明度将导致相分离或悬浮液,这将有助于操作问题。

Absorbance

通常,HPLC中使用的检测器基于样品竞争分物的光吸光度。选定波长范围内的流动相分子的固有吸光度不应干扰样品的吸光度。理想情况下,流动相溶剂应在感兴趣的波长下没有吸光度。

Volatility

流动相溶剂应具有低挥发性,特别是与光散射探测器一起使用。高挥发性溶剂可导致组成变化移动阶段composition over use and storage. This can lead to poor reproducibility of chromatograms.

Viscosity

所选择的溶剂应具有低粘度,使得流过柱不会导致高压压力的发展。

惯性

选择的溶剂应该惰性样本有限公司mponents, column packing and column material. Any reactivity with any of these components can lead to formation of precipitates, gases or other reaction products which can upset the system performance. The solvents should not form separate phase on coming in contact with the sample. In other words there should be complete miscibility of solvents.

水is also an important component of reverse phase mobile phase composition either in combination with other solvents or as buffer media. Water purification systems are commercially available. The water generated by such systems should be used at the earliest possible so that its quality does not degrade over the storage period.

HPLC技术对移动相的组成的变化高度敏感,从而达到检测器,因此必须通过0.45μm过滤器过滤溶剂以除去任何固体悬浮液。脱气也是去除任何可能导致流动限制或杂散峰的任何溶解空气的必要要求。

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