(M1430-08-45) A Systematic Approach to Analytical Method Transfer to Modern UHPLC Instruments

Purpose: Method life-cycle management is a regulatory requirement in the pharmaceutical industry. Two key tasks in method life-cycle management are performance monitoring and modernization. One pathway to modernization can be the transfer of existing methods to improved laboratory instrumentation. Analytical method transfers are documented processes that qualify a receiving laboratory to execute testing with an analytical method that originated in another laboratory. The method transfer process can also be applied to qualify new equipment by comparing chromatography results obtained between new and originating instruments. A mechanistic understanding of the instruments and test methods being qualified is critical in determining the suitability and optimization of the instrument and method. In this work, we will demonstrate how a leading CDMO has successfully transferred liquid chromatography methods onto a novel UHPLC platform.

Methods: Gradient delay volumes of the originator system and the target liquid chromatography system were assessed. In some cases, the exact configuration of the originator system was known, which facilitated the transfer onto the new UHPLC platform. In these cases, the pump and injection volume were taken into account to meet the originator system gradient delay volume. For example, a liquid chromatography system had a gradient delay volume of 1,100 μL and the method needed to be transferred onto a UHPLC equipped with a quaternary pump and a 100 μL injection loop, which had a default gradient delay volume of 974 μL, a gradient delay volume difference of 127 μL from the originator system. In many cases, this difference did not necessitate further modifications to the system configurations as the chromatography was meeting the method system suitability requirements. Should the chromatography differ, additional capillaries could be added. In cases where the exact configuration of the originator system was not known, the gradient delay volume was evaluated by running a gradient from 0% to 100% 10 mg/mL caffeine in water. The gradient delay volume (GDV) is then calculated by taking into account the flow rate (FR) in mL/min, time of 50% response (T50) and Time of Gradient excluding hold times (TG) as per the following equation, GDV = FR × (T50 – (0.5 × TG)).

Results: Approximately 200 UHPLC and HPLC methods ranging from R&D to commercial phase were transferred from several HPLC and UHPLC systems to the new UHPLC systems. The new systems offer improved sensitivity and reduced system suitability failures through reduced baseline fluctuation cause by both pump and injection pressure fluctuations. Examples of this can be seen in table 1 and 2 comparing the performance of the new system with other liquid chromatography platform for meeting system suitability. The transfer data indicated that the new UHPLC system provided a viable route to method modernization with minimal method re-optimization or validation.

Conclusion: Method transfer should not be cumbersome. With good knowledge of regulatory requirements, an understanding of how the target liquid chromatography system compares to the origin system, and with documented and characterized system modifications, method transfer should no longer be a bottleneck to modernization. More importantly, modernizing methods to a new UHPLC platform can lead to significant improvements to an analytical method, such as better resolution, improved sensitivity, and reduced system suitability failures.


Table 1: System Suitability Summary


Table 2: System Suitability Summary