Oasis 2x4 SPE Method DevelopmentBioanalysis, Metabolomics, Biopharma
┛This method showcases an SPE-based Sample Prep┏
Andrew+ Automated Mixed-Mode SPE Method Development
Quantitative and qualitative LC-MS analysis of complex biological samples requires extensive sample clean-up procedures to remove undesirable matrix components that may otherwise coelute and adversely affect the quantitation of the target analyte. Mixed-mode (reverse phase-ion exchange) solid-phase extraction (SPE) is an effective and well-proven technique for sample cleanup. However, developing mixed-mode SPE methods for the quantitation of analytes can be a tedious process. A good understanding of the physical and chemical properties of the analyte and the SPE sorbent chemistries is essential to selecting a suitable mixed-mode SPE sorbent that will not only remove the undesired components from the sample matrix but also result in good analyte recovery from the sample matrix. Furthermore, mixed-mode SPE method development involves several steps requiring precise and accurate pipetting of samples, reagents, and solvents.
Waters Oasis Method Development or sorbent selection plate simplifies this sorbent selection process, wherein it includes all four different mixed-mode ion exchange/reverse phase sorbent chemistries (MCX, MAX, WAX, and WCX) in a single plate format and utilizes just two optimized and proven protocols (2x4 sorbent selection method) to develop a mixed-mode SPE method (Figure 1). While the Oasis Method Development or sorbent selection plate with the 2x4 protocol simplifies the sorbent selection process, automation of this Oasis 2x4 sorbent selection protocol using the Andrew+ pipetting robot makes the entire SPE sorbent selection process rapid, reproducible, accurate, and robust.
◼️ PROTOCOL | Oasis 2x4 SPE Method Development – Andrew+
In this protocol, the automation of the Oasis 2x4 sorbent selection method using the Oasis Method Development 96-well μElution Plate is demonstrated by extracting four analytes Imipramine, 1-decanesulfonic acid, Ketoprofen, and Valethamate from spiked human blood plasma samples. Each of these analytes produces good analyte recovery, specific to the four Oasis mixed-mode sorbent chemistries MCX, WAX, MAX, and WCX respectively. All the steps in the SPE sorbent selection method development from loading, washing, and elution as well as controlling the vacuum of the SPE manifold are automated by the Andrew+ platform, except for changing the SPE sample collection plate in the vacuum manifold between washing and elution steps of the protocol. However, vacuum control or programming of the vacuum pump is available only when the IKA VACSTAR control vacuum pump is used. The SPE extracted samples are then quantified by LC-MS.
Figure 1: Overview of the conditions used in the Oasis 2x4 SPE Method Development.
Andrew+ System Components
- Microplate Vacuum+ | p/n 176004579
- IKA VACSTAR control vacuum pump | p/n 0020109375 for EU/CH, 0020109376 for the US, 0020109377 for the UK
Consumables & Other Systems
- Oasis method development 96-well μElution plate, 2 mg sorbent/well | p/n 186004475
- Waters QuanRecovery™ 700 μL 96-well plate | p/n 186009185
- Axygen 12-well reservoir, 12 channel trough | p/n RES-MW12-HP
- Falcon 50 mL conical centrifuge tube | p/n 352070
- Waters silicone/PTFE cap mat for 96-well plate, 7 mm round wells | p/n 186006333
- ACQUITY UPLC BEH C18 column, 130Å, 1.7 µm, 2.1x50 mm | p/n 176000863
- ACQUITY UPLC I-Class system
- Xevo TQ-S Tandem Quadrupole Mass Spectrometer