How Advances in Bioanalysis Drive Scientific and Ethical Progress

How Sygnature Discovery Supports the 3Rs with LC-MS/MS

Scientific Precision and Ethical Innovation

In the world of drug discovery, researchers face a critical challenge: how do you generate high-quality, reproducible bioanalytical data while minimizing the ethical and logistical burden of using animals in testing?  This is especially so with central nervous system (CNS) drug discovery, where obtaining biological samples from the CNS is not straightforward, resulting in multiple animals often being required to obtain the necessary data.

What are the 3Rs in Relation to Animal Use in Drug Discovery?

1. Reduction: Reducing the number of animals needed without compromising data
2. Refinement: Real-time, minimally invasive sampling with fewer ethical concessions
3. Replacement: Obtaining more insight per subject while reducing replication

Addressing the core tenets behind the 3 Rs is possible by combining advanced LC-MS/MS bioanalysis platforms and rethinking traditional pharmacokinetic/pharmacodynamic (PK/PD) study designs with new techniques like microdialysis.

From Compromise to Confidence: The 3Rs in Action

Historically, CNS PK/PD studies required larger sample volumes, leading to high animal use and associated variability. Sygnature challenged this model by integrating LC-MS/MS assays with low-volume microdialysis serial sampling techniques, dramatically reducing the number of animals needed without compromising data integrity.

1. Reduction: Less Animal-Based Testing, More Data

Using ultra-sensitive LC-MS/MS technology—notably the Waters™ Xevo™ TQ Absolute Mass Spectrometer, Sygnature reliably quantified neurotransmitters and drug compounds from as little as 10-20 μL of artificial CSF (aCSF). Their optimized PK workflow reduces animal usage from 24 animals down to just three animals per study, improving ethical compliance and improving data robustness with PK profiles generated for the same animal.  

“This has fundamentally reduced inter-animal variability while maximizing the data we get from each study.”
— Dr. Stuart Best, Deputy Director of Drug Metabolism and Pharmacokinetics, Sygnature Discovery

2. Refinement: Real-Time Monitoring with Microdialysis

Microdialysis enables real-time serial CNS sampling in conscious animals, eliminating the need for terminal single timepoint procedures. As LC-MS/MS technologies continue to become more sensitive, this approach, even with its low sample volumes, delivers insightful data.

3. Replacement: Smarter Study Design, Less Animal Dependence

Full replacement of animal models remains a long-term goal, but Sygnature’s approach shifts the paradigm. By extracting more insight per subject and reducing unnecessary replication, their team demonstrates that ethical, data-rich CNS research is not only possible— but scalable.

A New Era of CNS Bioanalysis: Supported by Waters

Sygnature Discovery’s approach highlights how cutting-edge technology enables both scientific and ethical progress. Leveraging Waters UPLC™ and tandem quadrupole MS systems, including the Xevo TQ Absolute and ACQUITY™ UPLC Systems, they’ve redefined what’s possible in low-volume, high-throughput CNS bioanalysis.

“Our ability to perform high-sensitivity CNS bioanalysis from such small volumes is a game-changer—for both the science and the animals.”
— Dr. Stuart Best

Why CNS Bioanalysis Matters

Sygnature Discovery’s integrated approach enables:

• High-throughput, high-sensitivity CNS bioanalysis
• Ethical animal studies aligned with the 3Rs
• Comprehensive PK/PD modelling from minimal sample volumes

“LC-MS/MS has transformed our ability to maximize bioanalytical data from the smallest possible samples. It’s invaluable for CNS discovery.”
— Dr. Stuart Best

Ready to Learn More?

 Dr. Stuart Best’s presentation is part of Navigating an Evolving Landscape in DMPK: Bioanalysis and Beyond- a webinar series from Waters that explores how the pharmaceutical industry is adapting to emerging modalities and ethical research practices through advanced bioanalytical technologies.

In this 30-minute on-demand session, “More from Less: LC-MS/MS – An Enabling Technology to Maximize Bioanalytical Data Collection from Small Sample Volumes”, Dr. Best shares how Sygnature Discovery is generating high-quality CNS data from low-volume samples while aligning with the 3Rs principles of ethical research.

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