The development of LC separations can be time-consuming and costly, with numerous steps, including literature research, column selection, method scouting development and optimization. To eliminate these steps, a software modeling tool that instantly models a separation on different column phases of preselected compounds was developed. Optimization can be performed while maintaining critical pair separations by adjusting for volume effects, mobile phases, gradient steps, and more. The modeler delivers a fast, no-cost starting point. The initial database includes a Drugs of Abuse (DoA) library and a Nitrosamines library with plans to continually expand the utility.
To build the modeler, a DoA library of 250 compounds was created. Retention times were collected using a fast/slow gradient, 30°C/60°C temperatures, and ACN/MeOH mobile phases on a single column dimension. Some additional data points outside of these runs were also collected for the development of a semiempirical correction factor that was used to improve modeling accuracy.
To assess the modeler’s accuracy, experiments comparing retention time values between wet-lab and modeled data were conducted. During development, the acceptance criteria for retention time agreement was set at +/- 15 seconds because it represents a typical MRM window. In the last stage of verification, only 13 retention time datapoints out of 704 collected exceeded the +/- 15 seconds window, giving a pass rate of 98.2%.
Once the initial library built, the modeler was evaluated over four increasingly more complex stages of verification. In the final stage, to assess the viability of adding future compounds to existing libraries, new compounds not present in the initial DoA library were added and then compared by testing different column dimensions and lengths, mobile phases, stationary phases, gradients programs, and temperatures against modeled retention time values.
This free virtual method development software can be used to deliver a fast, no-cost starting point for method development and optimization to LC users, no matter their expertise, via an on-demand consultative user experience that instantly generates a chromatogram and instrument-ready conditions (that can be further optimized by users) for the separation of preselected compounds.
