Simple and Efficient Solution for Robustness Testing in Gradient Elution Liquid Chromatographic Methods
Abstract
In this paper, an efficient way for robustness testing of gradient elution liquid chromatographic methods is proposed and tested on model mixtures comprising cilazapril, hydrochlorothiazide, and their degradation products, solutes that differ not only in polarities, but also in solubility and absorption characteristics. In general, the robustness could be tested with respect to various responses: resolution, retention factor, selectivity factor, change of detector response, etc. In chromatographic methods, the separation of the adjacent peaks is mandatory, and, consequently, the resolution is usually used as response. In isocratic elution methods, the resolution threshold depends on many factors, such as sizes of adjacent peaks, peak shapes, and asymmetry factor. At the same time, the situation is even more complex in gradient elution methods, because separation depends on a larger number of parameters, such as gradient profile, column geometry, mobile phase flow rate, column equilibra...tion between gradient runs, etc. To ensure baseline separation, the authors propose application of separation criterion (s) as response and indirect modeling in the robustness evaluation. Examined response in this approach is represented by the difference between the retention time of the beginning of the peak and the retention time of the end of the previously eluting peak of the critical pair. Moreover, the proposed methodology included reusing experiments from the optimization phase to define a robust chromatographic region without increasing the number of experiments. It was shown that method robustness can be easily and efficiently evaluated by this methodology.
Keywords:
Liquid chromatography / Robustness testing / Gradient elution / Separation criterion / Indirect modelingSource:
Chromatographia, 2018, 81, 8, 1135-1145Publisher:
- Springer Heidelberg, Heidelberg
Funding / projects:
DOI: 10.1007/s10337-018-3545-x
ISSN: 0009-5893
WoS: 000439334900002
Scopus: 2-s2.0-85047156435
Collections
Institution/Community
PharmacyTY - JOUR AU - Sljivić, Jasmina AU - Protić, Ana AU - Malenović, Anđelija AU - Otašević, Biljana AU - Zečević, Mira PY - 2018 UR - https://farfar.pharmacy.bg.ac.rs/handle/123456789/3208 AB - In this paper, an efficient way for robustness testing of gradient elution liquid chromatographic methods is proposed and tested on model mixtures comprising cilazapril, hydrochlorothiazide, and their degradation products, solutes that differ not only in polarities, but also in solubility and absorption characteristics. In general, the robustness could be tested with respect to various responses: resolution, retention factor, selectivity factor, change of detector response, etc. In chromatographic methods, the separation of the adjacent peaks is mandatory, and, consequently, the resolution is usually used as response. In isocratic elution methods, the resolution threshold depends on many factors, such as sizes of adjacent peaks, peak shapes, and asymmetry factor. At the same time, the situation is even more complex in gradient elution methods, because separation depends on a larger number of parameters, such as gradient profile, column geometry, mobile phase flow rate, column equilibration between gradient runs, etc. To ensure baseline separation, the authors propose application of separation criterion (s) as response and indirect modeling in the robustness evaluation. Examined response in this approach is represented by the difference between the retention time of the beginning of the peak and the retention time of the end of the previously eluting peak of the critical pair. Moreover, the proposed methodology included reusing experiments from the optimization phase to define a robust chromatographic region without increasing the number of experiments. It was shown that method robustness can be easily and efficiently evaluated by this methodology. PB - Springer Heidelberg, Heidelberg T2 - Chromatographia T1 - Simple and Efficient Solution for Robustness Testing in Gradient Elution Liquid Chromatographic Methods VL - 81 IS - 8 SP - 1135 EP - 1145 DO - 10.1007/s10337-018-3545-x ER -
@article{ author = "Sljivić, Jasmina and Protić, Ana and Malenović, Anđelija and Otašević, Biljana and Zečević, Mira", year = "2018", abstract = "In this paper, an efficient way for robustness testing of gradient elution liquid chromatographic methods is proposed and tested on model mixtures comprising cilazapril, hydrochlorothiazide, and their degradation products, solutes that differ not only in polarities, but also in solubility and absorption characteristics. In general, the robustness could be tested with respect to various responses: resolution, retention factor, selectivity factor, change of detector response, etc. In chromatographic methods, the separation of the adjacent peaks is mandatory, and, consequently, the resolution is usually used as response. In isocratic elution methods, the resolution threshold depends on many factors, such as sizes of adjacent peaks, peak shapes, and asymmetry factor. At the same time, the situation is even more complex in gradient elution methods, because separation depends on a larger number of parameters, such as gradient profile, column geometry, mobile phase flow rate, column equilibration between gradient runs, etc. To ensure baseline separation, the authors propose application of separation criterion (s) as response and indirect modeling in the robustness evaluation. Examined response in this approach is represented by the difference between the retention time of the beginning of the peak and the retention time of the end of the previously eluting peak of the critical pair. Moreover, the proposed methodology included reusing experiments from the optimization phase to define a robust chromatographic region without increasing the number of experiments. It was shown that method robustness can be easily and efficiently evaluated by this methodology.", publisher = "Springer Heidelberg, Heidelberg", journal = "Chromatographia", title = "Simple and Efficient Solution for Robustness Testing in Gradient Elution Liquid Chromatographic Methods", volume = "81", number = "8", pages = "1135-1145", doi = "10.1007/s10337-018-3545-x" }
Sljivić, J., Protić, A., Malenović, A., Otašević, B.,& Zečević, M.. (2018). Simple and Efficient Solution for Robustness Testing in Gradient Elution Liquid Chromatographic Methods. in Chromatographia Springer Heidelberg, Heidelberg., 81(8), 1135-1145. https://doi.org/10.1007/s10337-018-3545-x
Sljivić J, Protić A, Malenović A, Otašević B, Zečević M. Simple and Efficient Solution for Robustness Testing in Gradient Elution Liquid Chromatographic Methods. in Chromatographia. 2018;81(8):1135-1145. doi:10.1007/s10337-018-3545-x .
Sljivić, Jasmina, Protić, Ana, Malenović, Anđelija, Otašević, Biljana, Zečević, Mira, "Simple and Efficient Solution for Robustness Testing in Gradient Elution Liquid Chromatographic Methods" in Chromatographia, 81, no. 8 (2018):1135-1145, https://doi.org/10.1007/s10337-018-3545-x . .