SPE—通过固相萃取进行样品富集和纯化—waters.docxVIP

SPE——通过固相萃取进行样品富集和纯化 为何使用固相萃取（SPE）技术 1. 您需要从样品中去除特定干扰物，以免它们在目标分析物的检测和定量过程中影响实验结果。在此处所示的示例中，不适当的样品制备方案未能去除干扰物，导致提取物呈现出残留的黄色干扰物，色谱图中目标分析物与多个干扰峰发生了重叠。 ? ? ? 2. 您需要提高初始样品中目标分析物的浓度，以便所用的分析技术能够更轻松地对其进行检测和准确定量。如果目标分析物可被较强地保留，那么可能需要在SPE色谱柱上加载较大的样品量，随后仅以极小体积的洗脱液将此分析物洗脱下来，由此提高样品中分析物的浓度。 ? ? ? 3. 您需要去除样品中的干扰物（即使不可见），这些干扰物会在质谱检测中抑制目标分析物的信号。在此处的示例中，蛋白沉淀法无法去除血浆提取物中的磷脂，从而造成严重的离子抑制。优化的复合模式SPE方案可获取最纯净的提取物，并可在最大程度上降低离子抑制效应。 ? What is Solid-Phase Extraction (SPE)? Dont be confused by the term solid-phase extraction [SPE]. A typical SPE device has 50 times more separation power than a simple, single liquid-liquid extraction. SPE is actually column liquid-solid chromatography. Since SPEis?liquid chromatography [LC], its practice is governed by LC principles. A sample is introduced into a column or a cartridge device containing a bed of appropriate particles, or other form, of a chromatographic packing material [stationary phase]. Solvent [mobile phase] flows through the bed. By choosing an appropriate combination of mobile and stationary phases, sample components may pass directly through the column bed, or they may be selectively retained. Individual compounds in the sample each typically appear to travel at different speeds through the device. Using a weaker solvent causes them to move slowly and/or be strongly retained. A stronger solvent speeds up their passage through the bed and elutes the analyte(s) in a more concentrated volume. Elution from an SPE device is usually done by increasing the strength of the mobile phase in a series of discrete, rather than continuous, steps during which selected analytes or interferences are either fully retained or rapidly eluted-this variation of gradient elution called a?step gradient. Most commonly, SPE is practiced using miniature column or cartridge devices. An example is shown here. A mixture of three dyes is loaded onto the cartridge in a weak solvent, causing strong sample retention in a narrow band that appears?black?at the column inlet.