适用于质谱分析的在线固相萃取除盐方法

建立了一个简单、快速、有效的适用于质谱或液相色谱-质谱联用的在线固相萃取(SPE)高通量除盐方法。方法分为单柱和双柱模式,借助于包含双梯度泵(上样泵/分析泵)、自动进样器和配有十通切换阀的柱温箱的高效液相色谱系统,完成样品的自动化在线除盐。单柱模式通过上样泵实现在SPE柱上进样和除盐,被分析物则保留在SPE柱上;除盐完成后,通过阀切换利用分析泵洗脱富集在SPE柱上的被分析物。双柱模式则在单柱模式基础上增加了1根SPE柱,在色谱管理软件控制下2根SPE柱轮流工作,高效率完成样品的在线除盐。该方法在结合质谱分析蛋白质、多肽等领域具有较好的应用前景。

On-line solid phase extraction for desalting coupled with mass spectrometric analysis

The work shown here describes a simple, fast and effective on-line solid phase extraction (on-line SPE) method for facilitative high-throughput sample desalting before the detection of mass spectrometry(MS)or liquid chromatography-mass spectrometry(LC-MS). This method includes single SPE column mode and dual SPE column mode. It accomplishes the on-line desalting with an Ultimate 3000 HPLC system equipped with a dual pump system (loading pump/analytical pump), an autosampler, a column oven equipped with a 2p-10p valve, controlled by a chromatography data system. In the single SPE column mode, sample loading and desalting were performed on the SPE column using the loading pump. The analytes were retained on the SPE column, and the salt in the sample solution was flushed out of the SPE column. After desalting, the retained analytes were eluted from the SPE column with the analytical pump. In the dual SPE column mode, two same SPE columns were used. Firstly, the sample loading and desalting were performed on the SPE column 1. The analytes were retained and the salt was flushed out. After desalting on the SPE column 1, the sample loading and desalting were performed on the SPE column 2. Meanwhile, the retained analytes were eluted from the SPE column 1 with the analytical pump. As both of the processes on the SPE columns 1 and 2 described above completed, the sample loading and desalting on the SPE column 1, and the elution of the analytes from the SPE column 2 started again. As the SPE columns 1 and 2 worked in turn, the on-line SPE desalting system was efficient. The eluted analytes from the SPE columns may be determined by MS directly, or separated on the analytical column and then determined by MS/UV.