The novel approach towards gradient elution HPLC method development
Аутори
Milenković, MilanĐajić, Nevena
Krmar, Jovana
Rašević, Marija
Malenović, Anđelija
Otašević, Biljana
Protić, Ana
Конференцијски прилог (Објављена верзија)
Метаподаци
Приказ свих података о документуАпстракт
Introduction: Gradient elution HPLC finds its
purpose in simultaneous analyses of solutes
covering wide range of polarities. However, the
instrument related factors, especially dwell volume, are frequently responsible for fizzy transfer and
short life cycle of the gradient elution method.
Therefore, it is advisable to incorporate dwell
volume into the optimization stage and avoid
transfer related failures. The chemometric
approach would enable selection of optimal
chromatographic conditions for different HPLC
instruments. The aim of this study was to propose
and test this approach in gradient elution method’s
development.
Materials and Methods: The experiments were
carried out on three chromatographic systems
(UPLC, UHPLC and HPLC), while the separation
was achieved on Kinetex C18 Core-shell column
(100 mm × 2.1 mm, 2.6 μm particle size). Design
of experiments was constructed in Design-Expert
11.0. Indirect modeling, grid point search and
graphical presentations wer...e done in Matlab
7.10.0.
Results: Dabigatrane etexilate mesylate and nine
structurally related compounds were selected as
suitable model mixture due to its complexity and
polarity. Method development was supported with
experimental design methodology, Placket –
Burman for screening and D-optimal design for
optimization purposes. Dwell volumes were
included in the optimization phase and in this way
the same optimal chromatographic conditions for
all three instruments were selected. They included 10 mM ammonium acetate buffer
with pH set to 4.9 using acetic acid, and
acetonitrile. The components of the mobile phase
were pumped into chromatographic system with
flow rate of 400 μL min-1 in a linear gradient mode:
at 0 minutes 24% (v/v) acetonitrile and 76% (v/v) of
buffer solution, at 15 minutes 54% (v/v) acetonitrile
and 46% (v/v) buffer solution. At 16 minutes the
acetonitrile content was back to 24% (v/v) and 76%
(v/v) of buffer solution. The re-equilibration time
was set to 5 minutes. The examined
chromatographic region is graphically presented
and optimal conditions are noticed as the cross
sections (yellow dots). The method was validated
and confirmed its utility on all instruments.
Conclusions: The proposed methodology
demonstrated its ability to predict joint optimal
chromatographic conditions for instruments with different values of dwell volume. The potential was
confirmed on complex model mixture and
instruments significantly differing in dwell volume
values. In this way the gap between developing
and routine needs could be overwhelmed, followed
by facilitated transfer of methods.
Извор:
13th International Symposium on Pharmaceutical Sciences (ISOPS), June 22-25, 2021, Ankara, Turkey, 2021, 227-228Издавач:
- Ankara University Faculty of Pharmacy
Финансирање / пројекти:
- Министарство науке, технолошког развоја и иновација Републике Србије, институционално финансирање - 200161 (Универзитет у Београду, Фармацеутски факултет) (RS-MESTD-inst-2020-200161)
Напомена:
- ISOPS 13, Abstract Book
Институција/група
PharmacyTY - CONF AU - Milenković, Milan AU - Đajić, Nevena AU - Krmar, Jovana AU - Rašević, Marija AU - Malenović, Anđelija AU - Otašević, Biljana AU - Protić, Ana PY - 2021 UR - https://farfar.pharmacy.bg.ac.rs/handle/123456789/4696 AB - Introduction: Gradient elution HPLC finds its purpose in simultaneous analyses of solutes covering wide range of polarities. However, the instrument related factors, especially dwell volume, are frequently responsible for fizzy transfer and short life cycle of the gradient elution method. Therefore, it is advisable to incorporate dwell volume into the optimization stage and avoid transfer related failures. The chemometric approach would enable selection of optimal chromatographic conditions for different HPLC instruments. The aim of this study was to propose and test this approach in gradient elution method’s development. Materials and Methods: The experiments were carried out on three chromatographic systems (UPLC, UHPLC and HPLC), while the separation was achieved on Kinetex C18 Core-shell column (100 mm × 2.1 mm, 2.6 μm particle size). Design of experiments was constructed in Design-Expert 11.0. Indirect modeling, grid point search and graphical presentations were done in Matlab 7.10.0. Results: Dabigatrane etexilate mesylate and nine structurally related compounds were selected as suitable model mixture due to its complexity and polarity. Method development was supported with experimental design methodology, Placket – Burman for screening and D-optimal design for optimization purposes. Dwell volumes were included in the optimization phase and in this way the same optimal chromatographic conditions for all three instruments were selected. They included 10 mM ammonium acetate buffer with pH set to 4.9 using acetic acid, and acetonitrile. The components of the mobile phase were pumped into chromatographic system with flow rate of 400 μL min-1 in a linear gradient mode: at 0 minutes 24% (v/v) acetonitrile and 76% (v/v) of buffer solution, at 15 minutes 54% (v/v) acetonitrile and 46% (v/v) buffer solution. At 16 minutes the acetonitrile content was back to 24% (v/v) and 76% (v/v) of buffer solution. The re-equilibration time was set to 5 minutes. The examined chromatographic region is graphically presented and optimal conditions are noticed as the cross sections (yellow dots). The method was validated and confirmed its utility on all instruments. Conclusions: The proposed methodology demonstrated its ability to predict joint optimal chromatographic conditions for instruments with different values of dwell volume. The potential was confirmed on complex model mixture and instruments significantly differing in dwell volume values. In this way the gap between developing and routine needs could be overwhelmed, followed by facilitated transfer of methods. PB - Ankara University Faculty of Pharmacy C3 - 13th International Symposium on Pharmaceutical Sciences (ISOPS), June 22-25, 2021, Ankara, Turkey T1 - The novel approach towards gradient elution HPLC method development SP - 227 EP - 228 UR - https://hdl.handle.net/21.15107/rcub_farfar_4696 ER -
@conference{ author = "Milenković, Milan and Đajić, Nevena and Krmar, Jovana and Rašević, Marija and Malenović, Anđelija and Otašević, Biljana and Protić, Ana", year = "2021", abstract = "Introduction: Gradient elution HPLC finds its purpose in simultaneous analyses of solutes covering wide range of polarities. However, the instrument related factors, especially dwell volume, are frequently responsible for fizzy transfer and short life cycle of the gradient elution method. Therefore, it is advisable to incorporate dwell volume into the optimization stage and avoid transfer related failures. The chemometric approach would enable selection of optimal chromatographic conditions for different HPLC instruments. The aim of this study was to propose and test this approach in gradient elution method’s development. Materials and Methods: The experiments were carried out on three chromatographic systems (UPLC, UHPLC and HPLC), while the separation was achieved on Kinetex C18 Core-shell column (100 mm × 2.1 mm, 2.6 μm particle size). Design of experiments was constructed in Design-Expert 11.0. Indirect modeling, grid point search and graphical presentations were done in Matlab 7.10.0. Results: Dabigatrane etexilate mesylate and nine structurally related compounds were selected as suitable model mixture due to its complexity and polarity. Method development was supported with experimental design methodology, Placket – Burman for screening and D-optimal design for optimization purposes. Dwell volumes were included in the optimization phase and in this way the same optimal chromatographic conditions for all three instruments were selected. They included 10 mM ammonium acetate buffer with pH set to 4.9 using acetic acid, and acetonitrile. The components of the mobile phase were pumped into chromatographic system with flow rate of 400 μL min-1 in a linear gradient mode: at 0 minutes 24% (v/v) acetonitrile and 76% (v/v) of buffer solution, at 15 minutes 54% (v/v) acetonitrile and 46% (v/v) buffer solution. At 16 minutes the acetonitrile content was back to 24% (v/v) and 76% (v/v) of buffer solution. The re-equilibration time was set to 5 minutes. The examined chromatographic region is graphically presented and optimal conditions are noticed as the cross sections (yellow dots). The method was validated and confirmed its utility on all instruments. Conclusions: The proposed methodology demonstrated its ability to predict joint optimal chromatographic conditions for instruments with different values of dwell volume. The potential was confirmed on complex model mixture and instruments significantly differing in dwell volume values. In this way the gap between developing and routine needs could be overwhelmed, followed by facilitated transfer of methods.", publisher = "Ankara University Faculty of Pharmacy", journal = "13th International Symposium on Pharmaceutical Sciences (ISOPS), June 22-25, 2021, Ankara, Turkey", title = "The novel approach towards gradient elution HPLC method development", pages = "227-228", url = "https://hdl.handle.net/21.15107/rcub_farfar_4696" }
Milenković, M., Đajić, N., Krmar, J., Rašević, M., Malenović, A., Otašević, B.,& Protić, A.. (2021). The novel approach towards gradient elution HPLC method development. in 13th International Symposium on Pharmaceutical Sciences (ISOPS), June 22-25, 2021, Ankara, Turkey Ankara University Faculty of Pharmacy., 227-228. https://hdl.handle.net/21.15107/rcub_farfar_4696
Milenković M, Đajić N, Krmar J, Rašević M, Malenović A, Otašević B, Protić A. The novel approach towards gradient elution HPLC method development. in 13th International Symposium on Pharmaceutical Sciences (ISOPS), June 22-25, 2021, Ankara, Turkey. 2021;:227-228. https://hdl.handle.net/21.15107/rcub_farfar_4696 .
Milenković, Milan, Đajić, Nevena, Krmar, Jovana, Rašević, Marija, Malenović, Anđelija, Otašević, Biljana, Protić, Ana, "The novel approach towards gradient elution HPLC method development" in 13th International Symposium on Pharmaceutical Sciences (ISOPS), June 22-25, 2021, Ankara, Turkey (2021):227-228, https://hdl.handle.net/21.15107/rcub_farfar_4696 .