Optimization of mixed-mode HPLC method intended for analysis of adrenergic drugs
Конференцијски прилог (Објављена верзија)
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One of widely used drug group refers to drugs that act through adrenergic system. In following
research, the intent was to develop High Performance Liquid Chromatography (HPLC) method for
separation of adrenergic drugs. Therefore, selected mixture of analytes contained bisoprolol (BP),
fenoterole (FT), doxazosin (DOX), tetrahydrazoline (TH) and lofexidine (LOF). One of the
advantages of this method reflects in expanding knowledge about the analytical behaviour of
lofexidine, which is limited. Since all of selected analytes are basic in nature, mixed-mode column,
which includes reverse phase (RP) and weak cation exchange (WCX) interactions, was considered as
adequate for this HPLC analysis. The pronunciation one of two separation mechanisms depends on
mobile phase content and pH, and as a consequence has the potential of selectivity modulation [1].
Analyses were performed on HPLC Thermo Acclaim Mixed Mode WCX-1 (3 μm, 2.1 x 150 mm)
column. During screening phase, factors of si...gnificant influence on cationic analytes behaviour were
determinated. For that reason, column temperature (30-38°C) and mobile phase composition
parameters (acetonitrile content (30-50% (v/v)), pH (3.8-5.6) and ionic strength (20-40 mM) of
aqueous part of mobile phase) were selected for optimization. Design of Experiments (DoE) was used
for simultaneous optimization of selected factors. Experimental plan was in line with face-centered
Central Composite Design. Optimization goals were set in order to adequately separate all critical
peak pairs, and execute the analysis in reasonable time. Since selectivity in mixed-mode HPLC can
be governed through mobile phase content, selectivity factor (α) values of critical peak pairs were set
as optimization goals (αBP/FT, αTH/BP, αLOF/DOX > 1.2). The retention factor of last eluting analyte (kDOX)
was desired not to be greater than 10, in order to assure the rational analysis time. Experimental plan
and mathematical models were obtained with Design-Expert 7.0.0 software. Obtained models were
statistically evaluated (R2, pred. R2 and adj. R2 > 0.99). The most pronounced effect on followed
responses had the organic solvent content, whose increase lead to αTH/BP
enhancing, and had the
opposite effect on αBP/FT, αLOF/DOX and kDOX. Higher ionic strength corresponded to better separation
of BP from both FT and TH. This can be assigned to competitive behaviour between analytes and
ions present in mobile phase [2]. Time analysis shortening resulted as a consequence of higher values
of organic solvent, ionic strength and temperature. However, mobile phase pH had the opposite effect,
which can be related to more expressed cationic interactions and greater retention of basic analytes
[2].
Taking into account effects of all factors, and according to generated Derringer’s desirability function
for multiobjective decision making, the values of the factors that give the optimal responses are
selected to be 44% of acetonitrile, ionic strength 36 mM, temperature of 38°C and mobile phase pH
of 5.1.
Извор:
IMA-2021, 12 International Conference on Instrumental Methods of Analysis, Modern Trends and Applications, 20-23 September 2021, Virtual event, 2021, 119-119Издавач:
- Aristotle University of Thessaloniki
- National Technical University of Athens
Институција/група
PharmacyTY - CONF AU - Svrkota, Bojana AU - Krmar, Jovana AU - Protić, Ana AU - Zečević, Mira AU - Otašević, Biljana PY - 2021 UR - https://farfar.pharmacy.bg.ac.rs/handle/123456789/4697 AB - One of widely used drug group refers to drugs that act through adrenergic system. In following research, the intent was to develop High Performance Liquid Chromatography (HPLC) method for separation of adrenergic drugs. Therefore, selected mixture of analytes contained bisoprolol (BP), fenoterole (FT), doxazosin (DOX), tetrahydrazoline (TH) and lofexidine (LOF). One of the advantages of this method reflects in expanding knowledge about the analytical behaviour of lofexidine, which is limited. Since all of selected analytes are basic in nature, mixed-mode column, which includes reverse phase (RP) and weak cation exchange (WCX) interactions, was considered as adequate for this HPLC analysis. The pronunciation one of two separation mechanisms depends on mobile phase content and pH, and as a consequence has the potential of selectivity modulation [1]. Analyses were performed on HPLC Thermo Acclaim Mixed Mode WCX-1 (3 μm, 2.1 x 150 mm) column. During screening phase, factors of significant influence on cationic analytes behaviour were determinated. For that reason, column temperature (30-38°C) and mobile phase composition parameters (acetonitrile content (30-50% (v/v)), pH (3.8-5.6) and ionic strength (20-40 mM) of aqueous part of mobile phase) were selected for optimization. Design of Experiments (DoE) was used for simultaneous optimization of selected factors. Experimental plan was in line with face-centered Central Composite Design. Optimization goals were set in order to adequately separate all critical peak pairs, and execute the analysis in reasonable time. Since selectivity in mixed-mode HPLC can be governed through mobile phase content, selectivity factor (α) values of critical peak pairs were set as optimization goals (αBP/FT, αTH/BP, αLOF/DOX > 1.2). The retention factor of last eluting analyte (kDOX) was desired not to be greater than 10, in order to assure the rational analysis time. Experimental plan and mathematical models were obtained with Design-Expert 7.0.0 software. Obtained models were statistically evaluated (R2, pred. R2 and adj. R2 > 0.99). The most pronounced effect on followed responses had the organic solvent content, whose increase lead to αTH/BP enhancing, and had the opposite effect on αBP/FT, αLOF/DOX and kDOX. Higher ionic strength corresponded to better separation of BP from both FT and TH. This can be assigned to competitive behaviour between analytes and ions present in mobile phase [2]. Time analysis shortening resulted as a consequence of higher values of organic solvent, ionic strength and temperature. However, mobile phase pH had the opposite effect, which can be related to more expressed cationic interactions and greater retention of basic analytes [2]. Taking into account effects of all factors, and according to generated Derringer’s desirability function for multiobjective decision making, the values of the factors that give the optimal responses are selected to be 44% of acetonitrile, ionic strength 36 mM, temperature of 38°C and mobile phase pH of 5.1. PB - Aristotle University of Thessaloniki PB - National Technical University of Athens C3 - IMA-2021, 12 International Conference on Instrumental Methods of Analysis, Modern Trends and Applications, 20-23 September 2021, Virtual event T1 - Optimization of mixed-mode HPLC method intended for analysis of adrenergic drugs SP - 119 EP - 119 UR - https://hdl.handle.net/21.15107/rcub_farfar_4697 ER -
@conference{ author = "Svrkota, Bojana and Krmar, Jovana and Protić, Ana and Zečević, Mira and Otašević, Biljana", year = "2021", abstract = "One of widely used drug group refers to drugs that act through adrenergic system. In following research, the intent was to develop High Performance Liquid Chromatography (HPLC) method for separation of adrenergic drugs. Therefore, selected mixture of analytes contained bisoprolol (BP), fenoterole (FT), doxazosin (DOX), tetrahydrazoline (TH) and lofexidine (LOF). One of the advantages of this method reflects in expanding knowledge about the analytical behaviour of lofexidine, which is limited. Since all of selected analytes are basic in nature, mixed-mode column, which includes reverse phase (RP) and weak cation exchange (WCX) interactions, was considered as adequate for this HPLC analysis. The pronunciation one of two separation mechanisms depends on mobile phase content and pH, and as a consequence has the potential of selectivity modulation [1]. Analyses were performed on HPLC Thermo Acclaim Mixed Mode WCX-1 (3 μm, 2.1 x 150 mm) column. During screening phase, factors of significant influence on cationic analytes behaviour were determinated. For that reason, column temperature (30-38°C) and mobile phase composition parameters (acetonitrile content (30-50% (v/v)), pH (3.8-5.6) and ionic strength (20-40 mM) of aqueous part of mobile phase) were selected for optimization. Design of Experiments (DoE) was used for simultaneous optimization of selected factors. Experimental plan was in line with face-centered Central Composite Design. Optimization goals were set in order to adequately separate all critical peak pairs, and execute the analysis in reasonable time. Since selectivity in mixed-mode HPLC can be governed through mobile phase content, selectivity factor (α) values of critical peak pairs were set as optimization goals (αBP/FT, αTH/BP, αLOF/DOX > 1.2). The retention factor of last eluting analyte (kDOX) was desired not to be greater than 10, in order to assure the rational analysis time. Experimental plan and mathematical models were obtained with Design-Expert 7.0.0 software. Obtained models were statistically evaluated (R2, pred. R2 and adj. R2 > 0.99). The most pronounced effect on followed responses had the organic solvent content, whose increase lead to αTH/BP enhancing, and had the opposite effect on αBP/FT, αLOF/DOX and kDOX. Higher ionic strength corresponded to better separation of BP from both FT and TH. This can be assigned to competitive behaviour between analytes and ions present in mobile phase [2]. Time analysis shortening resulted as a consequence of higher values of organic solvent, ionic strength and temperature. However, mobile phase pH had the opposite effect, which can be related to more expressed cationic interactions and greater retention of basic analytes [2]. Taking into account effects of all factors, and according to generated Derringer’s desirability function for multiobjective decision making, the values of the factors that give the optimal responses are selected to be 44% of acetonitrile, ionic strength 36 mM, temperature of 38°C and mobile phase pH of 5.1.", publisher = "Aristotle University of Thessaloniki, National Technical University of Athens", journal = "IMA-2021, 12 International Conference on Instrumental Methods of Analysis, Modern Trends and Applications, 20-23 September 2021, Virtual event", title = "Optimization of mixed-mode HPLC method intended for analysis of adrenergic drugs", pages = "119-119", url = "https://hdl.handle.net/21.15107/rcub_farfar_4697" }
Svrkota, B., Krmar, J., Protić, A., Zečević, M.,& Otašević, B.. (2021). Optimization of mixed-mode HPLC method intended for analysis of adrenergic drugs. in IMA-2021, 12 International Conference on Instrumental Methods of Analysis, Modern Trends and Applications, 20-23 September 2021, Virtual event Aristotle University of Thessaloniki., 119-119. https://hdl.handle.net/21.15107/rcub_farfar_4697
Svrkota B, Krmar J, Protić A, Zečević M, Otašević B. Optimization of mixed-mode HPLC method intended for analysis of adrenergic drugs. in IMA-2021, 12 International Conference on Instrumental Methods of Analysis, Modern Trends and Applications, 20-23 September 2021, Virtual event. 2021;:119-119. https://hdl.handle.net/21.15107/rcub_farfar_4697 .
Svrkota, Bojana, Krmar, Jovana, Protić, Ana, Zečević, Mira, Otašević, Biljana, "Optimization of mixed-mode HPLC method intended for analysis of adrenergic drugs" in IMA-2021, 12 International Conference on Instrumental Methods of Analysis, Modern Trends and Applications, 20-23 September 2021, Virtual event (2021):119-119, https://hdl.handle.net/21.15107/rcub_farfar_4697 .