SPE固相萃取方法开发选择色谱模式
下表汇总了各种SPE模式，为方法开发工作的开展提供了一个良好的起点：
固相萃取选择色谱模式及吸附剂
| |
反相 |
正相 |
离子交换 |
| 分析物 |
中-低极性 |
低-高极性/中性 |
带电荷、可电离 |
| 分离机制 |
基于疏水性的分离 |
基于极性的分离 |
基于电荷的分离 |
| 样品基质 |
水溶液 |
非极性有机溶剂 |
水溶液/低离子强度 |
| SPE吸附剂的活化/平衡 |
1. 用极性有机溶剂得到的溶剂化物
2. 水 |
非极性有机溶剂 |
低离子强度缓冲液 |
| 初步冲洗步骤 |
水溶液/缓冲液 |
非极性有机溶剂 |
低离子强度缓冲液 |
| 洗脱步骤 |
增加极性有机溶剂的含量 |
增加混合有机溶剂的洗脱强度 |
更强的缓冲液——通过调节离子强度或pH值而中和电荷 |
| |
|
|
AX
[阴离子交换] |
CX
[阳离子交换] |
| 吸附剂官能团 |
C18, tC18, C8, tC2, CN, NH2, HLB, RDX, Rxn RP |
Silica, Alumina, Florisil, Diol, CN, NH2 |
Accell Plus QMA, NH2, SAX, MAX, WAX |
Accell Plus CM, SCX, MCX, WCX, Rxn CX |
| 吸附剂表面极性 |
低至中等 |
高至中等 |
高 |
高 |
| 典型溶剂的极性范围 |
高至中等 |
低至中等 |
高 |
高 |
| 典型的上样溶剂 |
水、低强度缓冲液 |
正己烷、氯仿、二氯甲烷 |
水、低强度缓冲液 |
水、低强度缓冲液 |
| 典型的洗脱溶剂 |
MeOH/水、CH3CN/水 |
乙酸乙酯、丙酮、CH3CN |
缓冲液、高离子强度的盐类，提高pH |
缓冲液、高离子强度的盐类，降低pH |
| 样品洗脱顺序 |
最大极性样品组分最先洗脱出来 |
最弱极性样品组分最先洗脱出来 |
最弱电离样品组分最先洗脱出来 |
最弱电离样品组分最先洗脱出来 |
| 为洗脱化合物而做的流动相溶剂改变 |
减弱溶剂极性 |
增强溶剂极性 |
增加离子强度或提高pH值 |
增加离子强度或降低pH值 |
This has been a brief introduction to sample enrichment and purification using solid-phase extraction [SPE]. The best way to start using SPE is to first learn what others have done with analytes and/or matrices similar to those of interest to you. You will find > 7,700 references to the use of SPE in the Resource Library on waters.com. Fill in the blank with a partial compound or matrix name in the following search phrase:
“Sep-Pak” OR “Oasis” AND ______*
NOTE: Rather than risk a spelling error, use an asterisk [*] with a root name for best results. Using this same search string, even more references [> 60,000] may be found on GOOGLE Scholar.
Further reading:
J.C. Arsenault and P.D. McDonald, Beginners Guide to Liquid Chromatography, Waters [2007]; Order P/N 715001531 on waters.com
P.D. McDonald and E.S.P. Bouvier, A Sample Preparation Primer and Guide to Solid-Phase Extraction Methods Development, Waters [2001] Search for WA20300 on waters.com
Waters, Purity by SPE [2008]; Search for 720001692en on waters.com
U.D. Neue, P.D. McDonald, Topics in Solid-Phase Extraction. Part 1. Ion Suppression in LC/MS Analysis: A Review. Strategies for its elimination by well-designed, multidimensional solid-phase extraction [SPE] protocols and methods for its quantitative assessment [2005]; Search for 720001273en on waters.com
