Automated Plate & Cartridge RP-SPE
This basic method provides the core methodology for translating a workflow into OneLab-executable script(s) as an attempt to fully or semi-automate a specific procedure. It demonstrates the benefits of automation and highlights OneLab capabilities and best practices to promote solution adoption, helping transition from manual to a more automated approach. It can be used alone or serves as a building block for a more complex workflow and is easily adaptable to users' requirements.
Overview
┛This method showcases a Core SPE Sample Prep┏
Fully Automated Solid Phase Extraction Sample Preparation using the Andrew+ Pipetting Robot configured with the Extraction+ Connected Device
Solid phase extraction (SPE) is a common extraction technique used in bioanalysis due to its ability to remove matrix interference, add selectivity and also provide the ability to concentrate samples for improved detection limits. While highly beneficial, SPE extraction is often perceived as complex due to the many protocol steps required to accurately and reliably isolate or purify the target analyte(s) from various biological matrices. Implementing fully automated SPE sample preparation greatly simplifies the analytical workflow, reduces human error, and improves analytical method reproducibility.
Figure 1: Graphical illustration of the automated SPE sample preparation and LC-MS analysis workflow using Oasis HLB SPE plates and cartridges and the Andrew+ Pipetting Robot configured with the Extraction+ connected device.
Extraction+ is a fully automatable SPE system solution used in combination with Andrew+, which eliminates the need for user intervention during SPE sample preparation and extraction. The Extraction+ device is comprised of two modules, an SPE manifold and a vacuum pump, both controlled via the OneLab software. An image of the Extraction+ configured with the Andrew+ is shown in Figure 2. Key features of Extraction+ include:
- Compatibility to perform SPE in both cartridge and plate formats,
- Liquid dispensing, collection, and evacuation into plates, tubes, or bottles,
- Flow-through waste collection,
- Full vacuum pressure control, defined within the OneLab protocol, for evacuation of the liquid during the SPE method execution (Figure 3).
Figure 2: This photo displays the Andrew+ Pipetting Robot configured with the Extraction+ system along with labware Dominos, Andrew Alliance Bluetooth electronic pipettes, and a microplate gripper on the tool stand. [5] Extraction+ includes the connected vacuum pump, flow-through waste container, the vacuum manifold with the collar, and the integrated collar lifter. Here in this photo, 6 cc SPE cartridges are loaded into a specific 12-position adaptor and placed onto the Extraction+ manifold.
Provided herein, are generic protocols for the reversed-phase SPE extraction of target analytes using Waters Oasis HLB 1, 3, and 6cc cartridges as well as macro and μElution 96-well plates. Protocols follow a simple 3-step SPE procedure (load, wash, elute).
Figure 3: Generic reversed-phase SPE procedure for Oasis cartridge and plate formats. Volumes (assuming 1:1 sample dilution/pre-treatment) used in the OneLab scripts are highlighted for each step. Values are based on the guidance provided in the Waters "SPE Method Development Wall Chart"(Waters, p/n 720001981en) and the "Oasis HLB Cartridges and 96-well Plates Care and Use Manual" (Waters, p/n 715000109).
◼️ PROTOCOL 1 | Automated Oasis HLB 1 cc Cartridge Reversed-Phase SPE | Andrew+ Configured with Extraction+
Figure 4: Andrew+ deck layout for Protocol #1 "Automated Oasis HLB 1 cc Cartridge Reversed-Phase SPE" using Andrew+ configured with Extraction+.
ORDERING INFORMATION
Andrew+ System Components: Dominos, Devices, Electronic Pipettes & Tips
➤ Automated Oasis HLB 1 cc Cartridge Reversed-Phase SPE Protocol – Andrew+
- 2x 250mL Duran Bottle Domino | p/n 186010193
- 1x Microtube Domino | p/n 186009601
- 2x Tip Insertion System Domino | p/n 186009612
- 1x 5mL Tip Rack Holder Domino | p/n 186009599
- Extraction+ Base Kit with Plate Gripper | p/n 176005201
- Extraction+ 1cc Cartridge Kit including 1x Collection Labware Rack Domino | p/n 176005204
- Andrew Alliance Bluetooth Electronic Pipette, 1-ch 300 μL | p/n 186009606
- Andrew Alliance Bluetooth Electronic Pipette, 1-ch 1000 μL | p/n 186009766
- Andrew Alliance Bluetooth Electronic Pipette, 1-ch 5000 μL | p/n 186009608
- Sartorius, Optifit Tips, 5-350 μL (x2) | p/n 700013297
- Sartorius, Optifit Tips, 10-1000 μL (x26) | p/n 700013298
- Sartorius, Optifit Tips, 100-5000 μL (x1) | p/n 700013291
Consumables
- DURAN® 250 mL clear glass laboratory bottle | p/n 218013651
- Eppendorf 2 mL Safe-Lock tube | p/n 0030120094
- Waters Oasis HLB 1 cc flangeless Vac cartridge, 30 mg sorbent, 30 μm | p/n 186001879 (NOTE: In the script, a placeholder cartridge, the Oasis HLB 1 cc Vac cartridge with flange, 30 mg sorbent, 30 μm is used. Cartridges with flange and flangeless cartridges work similarly)
- Waters 2 mL LC/GC screw-top vial | p/n 186000272C
◼️ PROTOCOL 2 | Automated Oasis HLB 3 cc Cartridge Reversed-Phase SPE | Andrew+ Configured with Extraction+
Figure 5: Andrew+ deck layout for Protocol #2 "Automated Oasis HLB 3 cc Cartridge Reversed-Phase SPE" using Andrew+ configured with Extraction+.
ORDERING INFORMATION
Andrew+ System Components: Dominos, Devices, Electronic Pipettes & Tips
➤ Automated Oasis HLB 3 cc Cartridge Reversed-Phase SPE Protocol – Andrew+
- 2x 250mL Duran Bottle Domino | p/n 186010193
- 1x Microtube Domino | p/n 186009601
- 2x Tip Insertion System Domino | p/n 186009612
- 1x 5mL Tip Rack Holder Domino | p/n 186009599
- Extraction+ Base Kit with Plate Gripper | p/n 176005201
- Extraction+ 3cc Cartridge Kit including 1x Collection Labware Rack Domino | p/n 176005205
- Andrew Alliance Bluetooth Electronic Pipette, 1-ch 300 μL | p/n 186009606
- Andrew Alliance Bluetooth Electronic Pipette, 1-ch 1000 μL | p/n 186009766
- Andrew Alliance Bluetooth Electronic Pipette, 1-ch 5000 μL | p/n 186009608
- Sartorius, Optifit Tips, 5-350 μL (x2) | p/n 700013297
- Sartorius, Optifit Tips, 10-1000 μL (x25) | p/n 700013298
- Sartorius, Optifit Tips, 100-5000 μL (x2) | p/n 700013291
Consumables
- DURAN® 250 mL clear glass laboratory bottle | p/n 218013651
- Eppendorf 2 mL Safe-Lock tube | p/n 0030120094
- Waters Oasis HLB 3 cc flangeless Vac cartridge, 60 mg sorbent, 30 μm | p/n 186001880
- Waters 2 mL LC/GC screw-top vial | p/n 186000272C
◼️ PROTOCOL 3 | Automated Oasis HLB 6 cc Cartridge Reversed-Phase SPE | Andrew+ Configured with Extraction+
Figure 6: Andrew+ deck layout for Protocol #3 "Automated Oasis HLB 6 cc Cartridge Reversed-Phase SPE" using Andrew+ configured with Extraction+.
ORDERING INFORMATION
Andrew+ System Components: Dominos, Devices, Electronic Pipettes & Tips
➤ Automated Oasis HLB 6 cc Cartridge Reversed-Phase SPE Protocol – Andrew+
- 2x 250mL Duran Bottle Domino | p/n 186010193
- 1x Microtube Domino | p/n 186009601
- 1x Tip Insertion System Domino | p/n 186009612
- 1x 5mL Tip Rack Holder Domino | p/n 186009599
- Extraction+ Base Kit with Plate Gripper | p/n 176005201
- Extraction+ 6cc Cartridge Kit including 1x Collection Labware Rack Domino | p/n 176005206
- Andrew Alliance Bluetooth Electronic Pipette, 1-ch 1000 μL | p/n 186009766
- Andrew Alliance Bluetooth Electronic Pipette, 1-ch 5000 μL | p/n 186009608
- Sartorius, Optifit Tips, 10-1000 μL (x2) | p/n 700013298
- Sartorius, Optifit Tips, 100-5000 μL (x26) | p/n 700013291
Consumables
- DURAN® 250 mL clear glass laboratory bottle | p/n 218013651
- Eppendorf 2 mL Safe-Lock tube | p/n 0030120094
- Waters Oasis HLB 6 cc Vac cartridge, 150 mg sorbent, 60 μm | p/n 186003379 (NOTE: In the script, a placeholder cartridge, the Oasis HLB 6 cc Vac cartridge, 150 mg sorbent, 30 μm, is used)
- VWR® 5 mL snap-cap centrifuge tube | p/n 10002-728
◼️ PROTOCOL 4 | Automated Oasis HLB 96-well Macro Plate Reversed-Phase SPE | Andrew+ Configured with Extraction+
Figure 7: Andrew+ deck layout for Protocol #4 "Automated Oasis HLB 96-well Macro Plate Reversed-Phase SPE" using Andrew+ configured with Extraction+.
ORDERING INFORMATION
Andrew+ System Components: Dominos, Devices, Electronic Pipettes & Tips
➤ Automated Oasis HLB 96-well Macro Plate Reversed-Phase SPE Protocol – Andrew+
- 2x Storage Plate Domino | p/n 186009596
- 4x Tip Insertion System Domino | p/n 186009612
- Extraction+ Base Kit with Plate Gripper | p/n 176005201
- Extraction+ Plate Kit including 1x Deepwell Microplate Domino and 1x Storage Plate Domino| p/n 176005203
- Andrew Alliance Bluetooth Electronic Pipette, 8-ch 300 μL | p/n 186009607
- Andrew Alliance Bluetooth Electronic Pipette, 8-ch 1200 μL | p/n 186009615
- Sartorius, Optifit Tips, 5-350 μL (x96) | p/n 700013297
- Sartorius, Optifit Tips, 50-1200 μL (x240) | p/n 700013300
Consumables
- Agilent 6-column reagent reservoir | p/n 201284-100
- Waters Oasis HLB 96-square well plate, 30 mg sorbent/well, 30 μm | p/n WAT058951
- Waters 800 μL 96-round well collection plate | p/n 186002481
- Waters 2 mL 96-square well collection plate, cut corner H1 | p/n 186002482
- Waters pre-slit PTFE/silicone cap mat for 96-well plate, round wells | p/n 186006332
◼️ PROTOCOL 5 | Automated Oasis HLB 96-well μElution Plate Reversed-Phase SPE | Andrew+ Configured with Extraction+
Figure 8: Andrew+ deck layout for Protocol #5 "Automated Oasis HLB 96-well μElution Plate Reversed-Phase SPE" using Andrew+ configured with Extraction+.
ORDERING INFORMATION
Andrew+ System Components: Dominos, Devices, Electronic Pipettes & Tips
➤ Automated Oasis HLB 96-well μElution Plate Reversed-Phase SPE Protocol – Andrew+
- 2x Storage Plate Domino | p/n 186009596
- 5x Tip Insertion System Domino | p/n 186009612
- Extraction+ Base Kit with Plate Gripper | p/n 176005201
- Extraction+ Plate Kit including 1x Deepwell Microplate Domino and 1x Storage Plate Domino | p/n 176005203
- Andrew Alliance Bluetooth Electronic Pipette, 8-ch 300 μL | p/n 186009607
- Andrew Alliance Bluetooth Electronic Pipette, 8-ch 1200 μL | p/n 186009615
- Sartorius, Optifit Tips, 5-350 μL (x200) | p/n 700013297
- Sartorius, Optifit Tips, 50-1200 μL (x120) | p/n 700013300
Consumables
- Agilent 6-column reagent reservoir | p/n 201284-100
- Waters Oasis HLB 96-well μElution plate, 2 mg sorbent/well, 30 μm | p/n 186001828BA
- Waters 800 μL 96-round well collection plate | p/n 186002481
- Waters 2 mL 96-square well collection plate, cut corner H1 | p/n 186002482
- Waters pre-slit PTFE/silicone cap mat for 96-well plate, round wells| p/n 186006332
Find more information about Recommended Pretreatment, Wash, and Elution Solutions in the:
Protocols
Contact info
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. 