Molnár-Institute collaborates on research on revolutionizing UHPLC with QbD principles

Berlin: – Scientists from the Molnár-Institute for Applied Chromatography have teamed up with Hungarian colleagues to author a significant research paper on new approaches to ultra-high performance liquid chromatography (UHPLC).

The study ‘Exploring better column selectivity choices in ultra-high performance liquid chromatography using Quality by Design principles’ is lead authored by Dr. Róbert Kormány from Budapest-based Egis Plc, working in collaboration with Molnár-Institute President and founder, Dr. Imre Molnár and colleague Dr. Hans-Jürgen Rieger.

Drastically reducing analysis time

Their research effort reworked an older method for amlodipine with the goal of reducing analysis time from one hour to under six minutes.

To select the best column for short and robust analysis, the team investigates nine different UHPLC column chemistries were investigated using 3-dimensional resolution spaces based on 12 experiments using modelling software.

The main variables used were gradient time (tG), temperature (T) and the pH of eluent A. The best critical resolution was calculated and located in a 3D space in an automated fashion and the corresponding best experiments were carried out. The work (9 × 12 = 108 runs) was finished with an UPLC instrument in less than 24 hours.

The comparison between predictions and real experiments showed an excellent correlation with differences typically less than 0.04 min (<3 s) in average, although the set points were located at quite different conditions on gradient times, pH’s and temperatures for the individual columns. All columns could perform the required baseline separation at their individual best working points with satisfactory results. However, method robustness was also depending on the column chemistry.

Conclusions

The team say that their method development for UHPLC separations according to QbD principles has demonstrated that if eluent properties are carefully considered with a wide variety of column chemistries, retention modelling can be used to find solutions for best separation for almost every column with analysis time reduced from 60 min to under a tenth of that.

“Although there are in terms of method robustness observable differences between the individual columns, the results are showing a much easier handling of the separation as such. This is a great advantage in the rapid development of the best possible separation in industrial units, helping to develop new drugs faster for many diseases, which could not be treated before,” the researchers write.

They note that their work breaks new ground in showing a scientifically reliable way of comparing columns in a highly variable multifactorial Design Space, reflecting not only column chemistries, but also the method robustness, which depends on the influence of gradient time, pH, ternary eluent composition, flow rate, starting and final percentage organic eluent composition at the same time.

“This is a new scientific approach, which enables us to gain insight of the fundamentals of multifactorial variabilities of UHPLC methods. Besides of the scientific importance of precise predictions of chromatograms, the paper is also important for the application of reliable science at reduced costs in the pharmaceutical industry, helping to update older pharmacopoeia methods,” the authors conclude.

About Molnar-Institute

Molnár-Institute for applied chromatography focuses on new strategies for improving the efficiency and success of modern high-performance liquid chromatography (HPLC) through its method development software DryLab^®4, HPLC and software training courses, and HPLC consulting.

Since its foundation in 1981, by Dr. Imre Molnár in Berlin, Molnár-Institute has followed a consistent mission, to form a center of excellence to save, reduce and optimize method development and run time in HPLC.

The institute supports ongoing research and development at universities and corporations worldwide with a team of chromatography specialists devoted to improving global health care and to ensuring the development of safe and effective products in the pharmaceutical, life-science, and food industries.

By applying its proprietary method development software DryLab®4 in conjunction with rigorous methodologies, the institute is able to optimize method development and reduce method run time in HPLC and UHPLC. Using real data from a small number of experimental runs, DryLab® builds multi-dimensional selectivity maps depicting the interaction of the most important parameters influencing an HPLC separation. This systematic, guided method development process can halve development time, help users optimize parameters based on their specific method goals, and enable easier compliance with regulatory standards (QbD).

Dr. Imre Molnár remains president of the Molnar-Institute, which is a privately-owned organisation, headquartered in central Berlin.

About Egis Plc

Egis Pharmaceuticals Plc, headquartered in Budapest, is a leading generic pharmaceutical company operating principally in Central and Eastern Europe and active in full pharmaceutical value chain from R&D through production of APIs and drug products to sales and marketing in more than 60 countries.

Egis has extensive experience in development of value-added generic molecules, efficient manufacturing processes, robust analytical methods for determination of impurities, comprehensive knowledge of organic chemical reactions and operations and strong IP support

Its recent investments have significantly increased development capabilities, especially with the construction of a new API R&D Centre including a dedicated lab for HPAPI development, headed by . Dr. Róbert Kormány.