Speed versus quality isn’t a new dilemma. The phrase, “Done fast, done cheaply, done right. You can pick two,” applies to many industries. And yet, the introduction of high throughput in vitro ADME assays with short turnaround times has played a major role in the drop of clinical attrition due to poor pharmacokinetics (PK) from 40% to 10% in the 1990s. But do these gains in speed come at a cost in data quality?
Dr. Graeme Clark, Head of Analytical at Cyprotex, says there are tradeoffs, pointing out that some Phase II conjugates can fragment back to the parent compound in atmospheric pressure ionisation sources, skewing the results of the metabolic stability assays. Without prior knowledge of the structure of the parent compound, it may not be flagged for this potential issue. However, high quality analytical data is within reach with a highly specialised and finely tuned system in place.
A robust LC-MS/MS platform lays the foundation for achieving high quality bioanalytical data using short run times. Generic protocols that can be optimised for a broad range of chemicals and assay matrices are a critical component. To determine potential regions of ion suppression and/or enhancement from common endogenous material, post-column infusion and LC-MS/MS methods should be applied. These evaluations should be performed across different HPLC stationary phases, mobile phases, additives and chromatographic gradients for a panel of compounds representative of drug-like molecules to allow establishment of a set of relevant generic LC-MS/MS methods. .
Further enhancements in throughput can be achieved using commercial software, however, to maintain quality, manual intervention and review is often necessary. Our internally built LIMS , known as Labsys™, has been designed to facilitate the compound optimisation process. A key advantage is that it can intelligently build automated sample lists for cassette analysis using parameters such as precursor ion, retention time, isotopic distribution and common biotransformations. LabsysTM allows ADME scientists to design an assay, prepare robotic scripts and generate LC-MS/MS sample lists and methods in parallel so that there is no risk of a mismatched method and cassette.
In summary, by using LabsysTM and commercial software in combination with a tiered LC-MS/MS system, we can deliver up to 4000 data points per system per overnight run with a >99% success rate for method optimisation and a >95% success rate for compound analysis from an assay, irrespective of matrix. The workflow ensures a highly efficient process and achieves a cost effective, high quality data in short turnaround. Now we can confidently say “Done fast, done cheaply, done right. You can pick three.”