Dilute and Shoot
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
Automated, Simple Dilute-and-Shoot Sample Preparation for Improved Accuracy and Superior Reproducibility
Ethyl glucuronide (EtG) and ethyl sulfate (EtS) are important biomarkers for monitoring alcohol use. Identification and quantification of EtG and EtS as biomarkers of ethanol use is performed for a wide range of testing purposes. Detecting these metabolites has proven beneficial as alcohol abuse is highly prevalent in many cultures and contributes considerably to the global burden of health and social issues. Therefore, there is a growing need for the detection and identification of ethanol use.
EtG and EtS are superior markers of alcohol intake due to a long detection window. EtG and EtS are minor water-soluble II metabolites of ethanol and are detectable in urine up to 80hrs following ethanol consumption. The sample preparation method detailed here is based upon the “dilute-and-shoot” approach. This method involves the dilution of samples with an internal standard before injection into the LC-MS system for the detection of target metabolites.
The dilute-and-shoot sample preparation can seem very straightforward compared to other analytical workflows. Nevertheless, repetitive pipetting can be tedious and error-prone especially when processing a large number of samples. Developing the EtG/EtS dilute and-shoot method on Andrew+ guarantees accurate pipetting and efficient mixing of samples. The automation of this workflow frees up analyst expertise for other tasks and requires minimal training to run. The OneLab cloud-native software guides the user when running a protocol with the required materials and deck setup. Andrew+ provides reproducible and consistent pipetting with every run, eliminating the risk of user error associated with manual operations.
Automation Benefits
- Mitigate the risk of error during sample preparation and maximize repeatability
- Ensure seamless method transferability
- Improve lab efficiency – Automated pipetting enables better utilization of analyst’s time
- OneLab software is easily operated via an intuitive interface for designing and executing protocols
- Andrew+ is versatile with multiple tools and device options to fit different needs while ensuring high-performance standards - A comparable mixing efficiency is obtained using the Andrew Alliance pipettes and the Microplate Shaker+
◼️ PROTOCOL 1 | Dilute & Shoot - Pipette Mixing | Andrew+
ORDERING INFORMATION
Andrew+ System Components: Dominos, Electronic Pipettes & Tips
- Deepwell Microplate Domino | p/n 186009597
- Microtube Domino | p/n 186009601
- Storage Plate Domino | p/n 186009596
- 2x Tip Insertion System Domino | p/n 186009612
- Andrew Alliance Bluetooth Electronic Pipette, 1-ch 300 μL | p/n 186009606
- Andrew Alliance Bluetooth Electronic Pipette, 8-ch 1200 µL | p/n 186009615
- Sartorius, Optifit Tips, 5-350 μL | p/n 700013297
- Sartorius, Optifit Tips, 50-1200 µL | p/n 700013300
Consumables & Other Systems
- Waters 2 mL 96-square well collection plate, cut corner H1 | p/n 186002482
- Agilent 6-column reagent reservoir | p/n 201284-100
- Fisherbrand™ Premium 1.5 mL microtube | p/n 11926955
- Acquity UPLC CSH Phenyl-Hexyl column | p/n 186005408
- ACQUITY UPLC-I-Class (FTN) system
- Xevo TQD Mass Spectrometer
◼️ PROTOCOL 2 | Dilute & Shoot - Microplate Shaking | Andrew+
ORDERING INFORMATION
Andrew+ System Components: Dominos, Devices, Electronic Pipettes & Tips
- Deepwell Microplate Domino | p/n 186009597
- Microtube Domino | p/n 186009601
- 2x Tip Insertion System Domino | p/n 186009612
- Microplate Shaker+ | p/n 176004577
- Andrew Alliance Bluetooth Electronic Pipette, 1-ch 300 μL | p/n 186009606
- Andrew Alliance Bluetooth Electronic Pipette, 8-ch 1200 µL | p/n 186009615
- Sartorius, Optifit Tips, 5-350 μL | p/n 700013297
- Sartorius, Optifit Tips, 50-1200 µL | p/n 700013300
Consumables & Other Systems
- Waters 2 mL 96-square well collection plate, cut corner H1 | p/n 186002482
- Agilent 6-column reagent reservoir | p/n 201284-100
- Fisherbrand™ Premium 1.5 mL microtube | p/n 11926955
- Acquity UPLC CSH Phenyl-Hexyl column | p/n 186005408
- ACQUITY UPLC-I-Class (FTN) system
- Xevo TQD Mass Spectrometer
Figure 1: Dilute-and-shoot workflow using Andrew+. On the left, the Andrew+ deck layout to run the protocol "Dilute & Shoot" with pipette mixing; On the right, the Andrew+ deck layout to run the protocol "Dilute & Shoot" with plate shaking using the connected Microplate Shaker+ (position 5).
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. 