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Oasis HLB SPE method development using the 20 bottle approach

Click & Execute Method

Bioanalysis, LCMS sample prep

Published by

Waters Corporation

Version 1, 3 Dec 2023 at 11:49 AM

Solid phase extraction (SPE) is a routinely used sample preparation technique in bioanalytical/DMPK laboratories, selectively capturing the analyte or drug target while removing biomatrix components, thus improving the sensitivity, selectivity, and robustness of the analytical method. SPE is often perceived as complex and labor-intensive, requiring lengthy method optimization to achieve high analyte recovery. The 20 bottle method optimization approach described in the Oasis HLB care & use manual provides an elegant way of optimizing the organic composition of the wash and elution solvents in a single experiment, significantly simplifying the method development process.

Use in OneLab

Please note that this is a “Click & Execute Method”

This advanced method offers an end-to-end solution developed around an advanced application, a specialized kit, or an analytical system. It delivers complete functional, ready-to-use protocols that are qualitatively and quantitatively assessed for consistency, executability, and repeatability, usually without needing to make any changes. It operates per batch size and is scalable to accommodate changing requirements.

Overview

Bioanalysis, or the analysis of drugs and their metabolites from biological fluids, is used in support of pharmaceutical drug discovery, development, and research. SPE is often used in bioanalytical/DMPK laboratories as a sample preparation technique to improve extraction recoveries, obtain cleaner samples as well as to concentrate samples without the need for evaporation (which may cause sample loss).

The critical steps of the SPE workflow that can be optimized include the wash and elution solvent composition. The Oasis HLB care & use manual (715000109EN) provides a starting protocol that provides reasonable recoveries for a diversity of analytes. Additionally, it also describes an elegant SPE experimental design (Figure 2) which allows scientists to optimize the composition of the wash and elution solutions in a single experiment, thereby simplifying Oasis HLB method optimization.

Figure 1: Visual representation of Oasis HLB 20 bottle method optimization approach from care and use manual

The 20-bottle SPE method optimization approach describes an elegant experimental design that varies the pH and the organic content of the elution solution to determine which condition provides the best probability of eluting the compound of interest from the sorbent bed. The elution solvent composition varies from 0-100% organic (generally methanol) with either 2% ammonium hydroxide or 2% formic acid. The acidic or basic nature of the elution solvent helps alter the pH, whereas the organic content helps dissociate analytes from the sorbent bed and improve solubility.

Additionally, because this experimental approach provides an elution profile for the analytes at different concentrations of organic solvent, the same data can also be used to determine the highest % organic that can be used as a wash solvent which will eliminate unwanted matrix components without losing any analyte from the SPE sorbent, further enhancing method selectivity. The final experiment for SPE method development usually involves testing the optimized wash and elution solvents to extract analytes spiked in the matrix of interest to determine recovery and matrix effects.

Assay notes

Figure 2: OneLab Protocol Visualization

Figure 3: Andrew+ OneLab deck set up for the Oasis HLB SPE method optimization using the 20 bottle method. Samples and reagents locations on Andrew+ deck: [1-4] Tips Insertion System Domino, [5] Extraction+, [6] Storage Plate domino, [7] Conical Microtube Domino, [8] 50 mL Centrifuge Tube Domino, [9-11] 8-Channel Reservoir Domino

Protocol specifications

Oasis HLB 20 Bottle Method Optimization:

Estimated time of execution: 4h 31m 15s
Hands-on time: 1m (last step before analysis)
Tip consumption:
52 x 5-120 μL tips
120 x 10-300 μL tips
23 x 50-1000 µL tips

Ordering information

Andrew+ System Components: Dominos, Devices, Electronic Pipettes & Tips

Andrew+ Pipetting Robot
OneLab software
4x Tip Insertion System Domino | p/n 186009612
Extraction+ Base Kit with Plate Gripper | p/n 176005201
Storage Plate Domino | p/n 186009596
Microtube Domino | p/n 186009601
50mL Conical Centrifuge Tube Domino | p/n 186009614
2x 8-Channel Pipette Reservoir Domino | p/n 186009613
Andrew Alliance Bluetooth Electronic Pipette, 1-ch 120 µL | p/n 186009765
Andrew Alliance Bluetooth Electronic Pipette, 1-ch 1000 µL | p/n 186009766
Andrew Alliance Bluetooth Electronic Pipette, 8-ch 300 µL | p/n 186009607
2× Pipette Adaptor Single Channel | p/n 186009590
1× Pipette Adaptor Multi-Channel | p/n 186009591

Recommended consumables

Oasis HLB 96-well µElution Plate, 2 mg Sorbent per Well, 30 µm | p/n 186001828BA
96-well Sample Collection plate, 700 µL Round well | p/n 186005837
Polypropylene Cap Mat Round Well for 96-well Plate | p/n 186002483
INTEGRA 10 mL multichannel reservoir | p/n 4332
Falcon® 50 mL conical centrifuge tube | p/n 352070
Fisherbrand™ Premium 1.5 mL microtube | p/n 11926955

References

Waters Care & Use Manual 715000109EN - Oasis HLB Cartridges and 96-Well Plates (https://www.waters.com/webassets/cms/support/docs/715000109.pdf)

This is an open access protocol distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.