TY  - JOUR
AU  - Antonić, Tamara
AU  - Ardalić, Daniela
AU  - Vladimirov, Sandra
AU  - Zeljković, Aleksandra
AU  - Vekić, Jelena
AU  - Mitrović, Marija
AU  - Ivanišević, Jasmina
AU  - Gojković, Tamara
AU  - Munjas, Jelena
AU  - Spasojević-Kalimanovska, Vesna
AU  - Miković, Željko
AU  - Stefanović, Aleksandra
PY  - 2023
UR  - https://farfar.pharmacy.bg.ac.rs/handle/123456789/4957
AB  - A specific feature of dyslipidemia in pregnancy is increased high-density lipoprotein (HDL) cholesterol concentration, which is probably associated with maternal endothelium protection. However, preeclampsia is most often associated with low HDL cholesterol, and the mechanisms behind this change are scarcely explored. We aimed to investigate changes in HDL metabolism in risky pregnancies and those complicated by late-onset preeclampsia. We analyze cholesterol synthesis (cholesterol precursors: desmosterol, 7-dehydrocholesterol, and lathosterol) and absorption markers (phytosterols: campesterol and β-sitosterol) within HDL particles (NCSHDL), the activities of principal modulators of HDL cholesterol’s content, and major HDL functional proteins levels in mid and late pregnancy. On the basis of the pregnancy outcome, participants were classified into the risk group (RG) (70 women) and the preeclampsia group (PG) (20 women). HDL cholesterol was lower in PG in the second trimester compared to RG (p < 0.05) and followed by lower levels of cholesterol absorption markers (p < 0.001 for campesterolHDL and p < 0.05 for β-sitosterolHDL). Lowering of HDL cholesterol between trimesters in RG (p < 0.05) was accompanied by a decrease in HDL phytosterol content (p < 0.001), apolipoprotein A-I (apoA-I) concentration (p < 0.05), and paraoxonase 1 (PON1) (p < 0.001), lecithin–cholesterol acyltransferase (LCAT) (p < 0.05), and cholesterol ester transfer protein (CETP) activities (p < 0.05). These longitudinal changes were absent in PG. Development of late-onset preeclampsia is preceded by the appearance of lower HDL cholesterol and NCSHDL in the second trimester. We propose that reduced capacity for intestinal HDL synthesis, decreased LCAT activity, and impaired capacity for HDL-mediated cholesterol efflux could be the contributing mechanisms resulting in lower HDL cholesterol.
PB  - MDPI
T2  - International Journal of Molecular Sciences
T1  - Cholesterol Metabolic Profiling of HDL in Women with Late-Onset Preeclampsia
VL  - 24
IS  - 14
DO  - 10.3390/ijms241411357
ER  - 

@article{
author = "Antonić, Tamara and Ardalić, Daniela and Vladimirov, Sandra and Zeljković, Aleksandra and Vekić, Jelena and Mitrović, Marija and Ivanišević, Jasmina and Gojković, Tamara and Munjas, Jelena and Spasojević-Kalimanovska, Vesna and Miković, Željko and Stefanović, Aleksandra",
year = "2023",
abstract = "A specific feature of dyslipidemia in pregnancy is increased high-density lipoprotein (HDL) cholesterol concentration, which is probably associated with maternal endothelium protection. However, preeclampsia is most often associated with low HDL cholesterol, and the mechanisms behind this change are scarcely explored. We aimed to investigate changes in HDL metabolism in risky pregnancies and those complicated by late-onset preeclampsia. We analyze cholesterol synthesis (cholesterol precursors: desmosterol, 7-dehydrocholesterol, and lathosterol) and absorption markers (phytosterols: campesterol and β-sitosterol) within HDL particles (NCSHDL), the activities of principal modulators of HDL cholesterol’s content, and major HDL functional proteins levels in mid and late pregnancy. On the basis of the pregnancy outcome, participants were classified into the risk group (RG) (70 women) and the preeclampsia group (PG) (20 women). HDL cholesterol was lower in PG in the second trimester compared to RG (p < 0.05) and followed by lower levels of cholesterol absorption markers (p < 0.001 for campesterolHDL and p < 0.05 for β-sitosterolHDL). Lowering of HDL cholesterol between trimesters in RG (p < 0.05) was accompanied by a decrease in HDL phytosterol content (p < 0.001), apolipoprotein A-I (apoA-I) concentration (p < 0.05), and paraoxonase 1 (PON1) (p < 0.001), lecithin–cholesterol acyltransferase (LCAT) (p < 0.05), and cholesterol ester transfer protein (CETP) activities (p < 0.05). These longitudinal changes were absent in PG. Development of late-onset preeclampsia is preceded by the appearance of lower HDL cholesterol and NCSHDL in the second trimester. We propose that reduced capacity for intestinal HDL synthesis, decreased LCAT activity, and impaired capacity for HDL-mediated cholesterol efflux could be the contributing mechanisms resulting in lower HDL cholesterol.",
publisher = "MDPI",
journal = "International Journal of Molecular Sciences",
title = "Cholesterol Metabolic Profiling of HDL in Women with Late-Onset Preeclampsia",
volume = "24",
number = "14",
doi = "10.3390/ijms241411357"
}

Antonić, T., Ardalić, D., Vladimirov, S., Zeljković, A., Vekić, J., Mitrović, M., Ivanišević, J., Gojković, T., Munjas, J., Spasojević-Kalimanovska, V., Miković, Ž.,& Stefanović, A.. (2023). Cholesterol Metabolic Profiling of HDL in Women with Late-Onset Preeclampsia. in International Journal of Molecular Sciences
MDPI., 24(14).
https://doi.org/10.3390/ijms241411357

Antonić T, Ardalić D, Vladimirov S, Zeljković A, Vekić J, Mitrović M, Ivanišević J, Gojković T, Munjas J, Spasojević-Kalimanovska V, Miković Ž, Stefanović A. Cholesterol Metabolic Profiling of HDL in Women with Late-Onset Preeclampsia. in International Journal of Molecular Sciences. 2023;24(14).
doi:10.3390/ijms241411357 .

Antonić, Tamara, Ardalić, Daniela, Vladimirov, Sandra, Zeljković, Aleksandra, Vekić, Jelena, Mitrović, Marija, Ivanišević, Jasmina, Gojković, Tamara, Munjas, Jelena, Spasojević-Kalimanovska, Vesna, Miković, Željko, Stefanović, Aleksandra, "Cholesterol Metabolic Profiling of HDL in Women with Late-Onset Preeclampsia" in International Journal of Molecular Sciences, 24, no. 14 (2023),
https://doi.org/10.3390/ijms241411357 . .

TY  - JOUR
AU  - Rašević, Marija
AU  - Malenović, Anđelija
AU  - Protić, Ana
AU  - Zečević, Mira
PY  - 2023
UR  - https://farfar.pharmacy.bg.ac.rs/handle/123456789/4981
AB  - In this paper, method for enantiomeric purity testing of fourth-generation fluoroquinolone, moxifloxacin hydrochloride, was developed and validated. Exceptional enantioselectivity for this assay was achieved using cyclodextrin type Chiral Stationary Phase (CSP), phenylcarbamate-β-cyclodextrin CSP, and mobile phase consisted of acetonitrile and triethylammonium acetate (TEAA) buffer. Analytical Quality by Design (AQbD) methodology, comprising Design of Experiments (DoE) - Design Space (DS) approach, was used for method development. In order to select appropriate Critical Method Parameters (CMPs), risk assessment was done using combined three step strategy that involved Ishikawa diagram - CNX (Control, Noise and eXperimental) - FMEA (Failure Mode and Effect Analysis). Three CMPs were further selected and investigated in this study: acetonitrile content in the mobile phase (30–50%, v/v), triethylamine content in the TEAA buffer (0.1–1.5%, v/v) and aqueous phase pH (3.5–4.5). Monte Carlo simulations were performed and 3D-DS was computed. One point situated in the center of 3D-DS was selected as working point for method validation, with the following values of CMPs: acetonitrile content in the mobile phase set to 37% (v/v), triethylamine content in TEAA 0.8% and pH value of the aqueous phase set at 4.0. Also, 2D-DS was created (with fixed one factor – pH value of aqueous phase at 4.0) which also gave us confirmation that the selection of working conditions was suitable. The proposed enantioselective method was further on tested for its quantitative robustness, as well as for its suitability for the intended purpose through validation studies.
PB  - Elsevier B.V.
T2  - Journal of Pharmaceutical and Biomedical Analysis
T1  - Analytical method development supported by DoE-DS approach for enantioseparation of (S,S)- and (R,R)-moxifloxacin
VL  - 235
DO  - 10.1016/j.jpba.2023.115645
ER  - 

@article{
author = "Rašević, Marija and Malenović, Anđelija and Protić, Ana and Zečević, Mira",
year = "2023",
abstract = "In this paper, method for enantiomeric purity testing of fourth-generation fluoroquinolone, moxifloxacin hydrochloride, was developed and validated. Exceptional enantioselectivity for this assay was achieved using cyclodextrin type Chiral Stationary Phase (CSP), phenylcarbamate-β-cyclodextrin CSP, and mobile phase consisted of acetonitrile and triethylammonium acetate (TEAA) buffer. Analytical Quality by Design (AQbD) methodology, comprising Design of Experiments (DoE) - Design Space (DS) approach, was used for method development. In order to select appropriate Critical Method Parameters (CMPs), risk assessment was done using combined three step strategy that involved Ishikawa diagram - CNX (Control, Noise and eXperimental) - FMEA (Failure Mode and Effect Analysis). Three CMPs were further selected and investigated in this study: acetonitrile content in the mobile phase (30–50%, v/v), triethylamine content in the TEAA buffer (0.1–1.5%, v/v) and aqueous phase pH (3.5–4.5). Monte Carlo simulations were performed and 3D-DS was computed. One point situated in the center of 3D-DS was selected as working point for method validation, with the following values of CMPs: acetonitrile content in the mobile phase set to 37% (v/v), triethylamine content in TEAA 0.8% and pH value of the aqueous phase set at 4.0. Also, 2D-DS was created (with fixed one factor – pH value of aqueous phase at 4.0) which also gave us confirmation that the selection of working conditions was suitable. The proposed enantioselective method was further on tested for its quantitative robustness, as well as for its suitability for the intended purpose through validation studies.",
publisher = "Elsevier B.V.",
journal = "Journal of Pharmaceutical and Biomedical Analysis",
title = "Analytical method development supported by DoE-DS approach for enantioseparation of (S,S)- and (R,R)-moxifloxacin",
volume = "235",
doi = "10.1016/j.jpba.2023.115645"
}

Rašević, M., Malenović, A., Protić, A.,& Zečević, M.. (2023). Analytical method development supported by DoE-DS approach for enantioseparation of (S,S)- and (R,R)-moxifloxacin. in Journal of Pharmaceutical and Biomedical Analysis
Elsevier B.V.., 235.
https://doi.org/10.1016/j.jpba.2023.115645

Rašević M, Malenović A, Protić A, Zečević M. Analytical method development supported by DoE-DS approach for enantioseparation of (S,S)- and (R,R)-moxifloxacin. in Journal of Pharmaceutical and Biomedical Analysis. 2023;235.
doi:10.1016/j.jpba.2023.115645 .

Rašević, Marija, Malenović, Anđelija, Protić, Ana, Zečević, Mira, "Analytical method development supported by DoE-DS approach for enantioseparation of (S,S)- and (R,R)-moxifloxacin" in Journal of Pharmaceutical and Biomedical Analysis, 235 (2023),
https://doi.org/10.1016/j.jpba.2023.115645 . .

TY  - JOUR
AU  - Salkić, Alma
AU  - Otašević, Biljana
AU  - Rašević, Marija
AU  - Zečević, Mira
PY  - 2023
UR  - https://farfar.pharmacy.bg.ac.rs/handle/123456789/5143
AB  - A selective eco-friendly micellar HPLC method was developed for investigation of moxifloxacin and related compounds in the presence of its degradation products. Central composite design was used to optimize the experimental conditions. The proposed method is based on isocratic elution on a C18 column using 92.5% (v/v) biodegradable aqueous mobile phase containing 0.01 M sodium dihydrogen phosphate, 0.15 M sodium dodecyl sulfate (SDS) and 0.5% triethylamine (v/v) with a pH of 3.5 and 7.5% isopropanol (v/v) as eco- friendly organic solvent. The flow rate and injection volume were 0.6 ml/min and 5 µl, respectively. Experiments were performed at a temperature of 60 °C and detection was performed at 295 nm. The optimized method was validated. The method was found to be suitable for the quantification of moxifloxacin and its related compounds in moxifloxacin drug substance. The Green Analytical Procedure Index (GAPI) proves the superiority of the developed method against other reported methods.
PB  - Slovensko Kemijsko Drustvo
T2  - Acta Chimica Slovenica
T1  - Micellar Liquid Chromatographic Method for Determination of Moxifloxacin and its Impurities
VL  - 70
IS  - 3
SP  - 385
EP  - 397
DO  - 10.17344/acsi.2023.8022
ER  - 

@article{
author = "Salkić, Alma and Otašević, Biljana and Rašević, Marija and Zečević, Mira",
year = "2023",
abstract = "A selective eco-friendly micellar HPLC method was developed for investigation of moxifloxacin and related compounds in the presence of its degradation products. Central composite design was used to optimize the experimental conditions. The proposed method is based on isocratic elution on a C18 column using 92.5% (v/v) biodegradable aqueous mobile phase containing 0.01 M sodium dihydrogen phosphate, 0.15 M sodium dodecyl sulfate (SDS) and 0.5% triethylamine (v/v) with a pH of 3.5 and 7.5% isopropanol (v/v) as eco- friendly organic solvent. The flow rate and injection volume were 0.6 ml/min and 5 µl, respectively. Experiments were performed at a temperature of 60 °C and detection was performed at 295 nm. The optimized method was validated. The method was found to be suitable for the quantification of moxifloxacin and its related compounds in moxifloxacin drug substance. The Green Analytical Procedure Index (GAPI) proves the superiority of the developed method against other reported methods.",
publisher = "Slovensko Kemijsko Drustvo",
journal = "Acta Chimica Slovenica",
title = "Micellar Liquid Chromatographic Method for Determination of Moxifloxacin and its Impurities",
volume = "70",
number = "3",
pages = "385-397",
doi = "10.17344/acsi.2023.8022"
}

Salkić, A., Otašević, B., Rašević, M.,& Zečević, M.. (2023). Micellar Liquid Chromatographic Method for Determination of Moxifloxacin and its Impurities. in Acta Chimica Slovenica
Slovensko Kemijsko Drustvo., 70(3), 385-397.
https://doi.org/10.17344/acsi.2023.8022

Salkić A, Otašević B, Rašević M, Zečević M. Micellar Liquid Chromatographic Method for Determination of Moxifloxacin and its Impurities. in Acta Chimica Slovenica. 2023;70(3):385-397.
doi:10.17344/acsi.2023.8022 .

Salkić, Alma, Otašević, Biljana, Rašević, Marija, Zečević, Mira, "Micellar Liquid Chromatographic Method for Determination of Moxifloxacin and its Impurities" in Acta Chimica Slovenica, 70, no. 3 (2023):385-397,
https://doi.org/10.17344/acsi.2023.8022 . .

TY  - JOUR
AU  - Rmandić, Milena
AU  - Vasilić, Đorđe
AU  - Rašević, Marija
AU  - Zečević, Mira
AU  - Otašević, Biljana
AU  - Protić, Ana
AU  - Malenović, Anđelija
PY  - 2023
UR  - https://farfar.pharmacy.bg.ac.rs/handle/123456789/5055
AB  - In this study, an AQbD-compliant chaotropic chromatography method for ziprasidone and the determination of its five impurities was developed. The influence of critical method parameters (initial and final methanol fraction in the mobile phase, gradient duration) on the set of selected critical method attributes (t_imp. V, t_imp. V − t_imp. I, S and <WUSP>) was studied by Box–Behnken design. The errors resulting from the calculation of the model coefficients were propagated to the selected responses by Monte Carlo simulations, and their predictive distribution was obtained. The design space was computed (π ≥ 80%), and a working point was selected: initial methanol fraction 38.5%, final methanol fraction 77.5%, and gradient duration 16.25 min. Furthermore, the quantitative robustness of the developed method was tested using the Plackett–Burman design. P_imp II and P_imp V were found to be significantly affected, the first by mobile phase flow rate and the second by gradient duration. Finally, the method was validated, and its reliability for routine quality control in capsules was confirmed.
PB  - MDPI
T2  - Pharmaceuticals
T1  - Development of Analytical Quality by Design Compliant Chaotropic Chromatography Method for Ziprasidone and Its Five Impurities Determination
VL  - 16
IS  - 9
DO  - 10.3390/ph16091296
ER  - 

@article{
author = "Rmandić, Milena and Vasilić, Đorđe and Rašević, Marija and Zečević, Mira and Otašević, Biljana and Protić, Ana and Malenović, Anđelija",
year = "2023",
abstract = "In this study, an AQbD-compliant chaotropic chromatography method for ziprasidone and the determination of its five impurities was developed. The influence of critical method parameters (initial and final methanol fraction in the mobile phase, gradient duration) on the set of selected critical method attributes (t_imp. V, t_imp. V − t_imp. I, S and <WUSP>) was studied by Box–Behnken design. The errors resulting from the calculation of the model coefficients were propagated to the selected responses by Monte Carlo simulations, and their predictive distribution was obtained. The design space was computed (π ≥ 80%), and a working point was selected: initial methanol fraction 38.5%, final methanol fraction 77.5%, and gradient duration 16.25 min. Furthermore, the quantitative robustness of the developed method was tested using the Plackett–Burman design. P_imp II and P_imp V were found to be significantly affected, the first by mobile phase flow rate and the second by gradient duration. Finally, the method was validated, and its reliability for routine quality control in capsules was confirmed.",
publisher = "MDPI",
journal = "Pharmaceuticals",
title = "Development of Analytical Quality by Design Compliant Chaotropic Chromatography Method for Ziprasidone and Its Five Impurities Determination",
volume = "16",
number = "9",
doi = "10.3390/ph16091296"
}

Rmandić, M., Vasilić, Đ., Rašević, M., Zečević, M., Otašević, B., Protić, A.,& Malenović, A.. (2023). Development of Analytical Quality by Design Compliant Chaotropic Chromatography Method for Ziprasidone and Its Five Impurities Determination. in Pharmaceuticals
MDPI., 16(9).
https://doi.org/10.3390/ph16091296

Rmandić M, Vasilić Đ, Rašević M, Zečević M, Otašević B, Protić A, Malenović A. Development of Analytical Quality by Design Compliant Chaotropic Chromatography Method for Ziprasidone and Its Five Impurities Determination. in Pharmaceuticals. 2023;16(9).
doi:10.3390/ph16091296 .

Rmandić, Milena, Vasilić, Đorđe, Rašević, Marija, Zečević, Mira, Otašević, Biljana, Protić, Ana, Malenović, Anđelija, "Development of Analytical Quality by Design Compliant Chaotropic Chromatography Method for Ziprasidone and Its Five Impurities Determination" in Pharmaceuticals, 16, no. 9 (2023),
https://doi.org/10.3390/ph16091296 . .

TY  - JOUR
AU  - Milenković, Milan
AU  - Rašević, Marija
AU  - Otašević, Biljana
AU  - Zečević, Mira
AU  - Malenović, Anđelija
AU  - Protić, Ana
PY  - 2022
UR  - https://farfar.pharmacy.bg.ac.rs/handle/123456789/3977
AB  - Nowadays, method development is strongly focused on reducing time needed for method development and execution. This subject specially concerns gradient elution methods regarding the usual need for trouble shooting assistance with uncertain outcome during the method transfer from one laboratory to another. One of the main reasons for this situation is the dwell volume difference between HPLC systems. Therefore, the aim of this study was to propose a novel method development methodology that would integrate the dwell volumes differences in the optimization process. The proposed approach could be quite useful in industry that has insight in HPLC instruments planned to be used during the method life cycle. It was tested on the model mixture consisting of dabigatran etexilate mesylate and its nine impurities by use of perimental design methodology. Three different (U)HPLC instruments with high dwell volume differences were selected to challenge the methodology. Plan of experiments was defined with Plackett-Burman design for screening phase and D-optimal design for optimization phase. Initial and final amount of organic modifier, time of the gradient elution and pH value of the aqueous phase were selected as variables nificant for the gradient programme profile and included in the optimization stage along with dwell volume values. The separation criteria s between critical peak pairs was selected as output for method optimization while indirect modelling together with Monte Carlo simulations enabled selection of optimal and robust chromatographic conditions. They included 24% (v/v) of initial amount of acetonitrile, 54% (v/v) of the final amount of acetonitrile, 15 min of gradient elution run time and pH value equal to 4.9. The proposed method was successfully validated, met all validation criteria and thus proved its utility.
PB  - Elsevier B.V.
T2  - Journal of Pharmaceutical and Biomedical Analysis
T1  - Generic approach in a gradient elution HPLC method development that enables troubleshooting free method transfer
VL  - 207
DO  - 10.1016/j.jpba.2021.114367
ER  - 

@article{
author = "Milenković, Milan and Rašević, Marija and Otašević, Biljana and Zečević, Mira and Malenović, Anđelija and Protić, Ana",
year = "2022",
abstract = "Nowadays, method development is strongly focused on reducing time needed for method development and execution. This subject specially concerns gradient elution methods regarding the usual need for trouble shooting assistance with uncertain outcome during the method transfer from one laboratory to another. One of the main reasons for this situation is the dwell volume difference between HPLC systems. Therefore, the aim of this study was to propose a novel method development methodology that would integrate the dwell volumes differences in the optimization process. The proposed approach could be quite useful in industry that has insight in HPLC instruments planned to be used during the method life cycle. It was tested on the model mixture consisting of dabigatran etexilate mesylate and its nine impurities by use of perimental design methodology. Three different (U)HPLC instruments with high dwell volume differences were selected to challenge the methodology. Plan of experiments was defined with Plackett-Burman design for screening phase and D-optimal design for optimization phase. Initial and final amount of organic modifier, time of the gradient elution and pH value of the aqueous phase were selected as variables nificant for the gradient programme profile and included in the optimization stage along with dwell volume values. The separation criteria s between critical peak pairs was selected as output for method optimization while indirect modelling together with Monte Carlo simulations enabled selection of optimal and robust chromatographic conditions. They included 24% (v/v) of initial amount of acetonitrile, 54% (v/v) of the final amount of acetonitrile, 15 min of gradient elution run time and pH value equal to 4.9. The proposed method was successfully validated, met all validation criteria and thus proved its utility.",
publisher = "Elsevier B.V.",
journal = "Journal of Pharmaceutical and Biomedical Analysis",
title = "Generic approach in a gradient elution HPLC method development that enables troubleshooting free method transfer",
volume = "207",
doi = "10.1016/j.jpba.2021.114367"
}

Milenković, M., Rašević, M., Otašević, B., Zečević, M., Malenović, A.,& Protić, A.. (2022). Generic approach in a gradient elution HPLC method development that enables troubleshooting free method transfer. in Journal of Pharmaceutical and Biomedical Analysis
Elsevier B.V.., 207.
https://doi.org/10.1016/j.jpba.2021.114367

Milenković M, Rašević M, Otašević B, Zečević M, Malenović A, Protić A. Generic approach in a gradient elution HPLC method development that enables troubleshooting free method transfer. in Journal of Pharmaceutical and Biomedical Analysis. 2022;207.
doi:10.1016/j.jpba.2021.114367 .

Milenković, Milan, Rašević, Marija, Otašević, Biljana, Zečević, Mira, Malenović, Anđelija, Protić, Ana, "Generic approach in a gradient elution HPLC method development that enables troubleshooting free method transfer" in Journal of Pharmaceutical and Biomedical Analysis, 207 (2022),
https://doi.org/10.1016/j.jpba.2021.114367 . .

TY  - JOUR
AU  - Đajić, Nevena
AU  - Krmar, Jovana
AU  - Rmandić, Milena
AU  - Rašević, Marija
AU  - Otašević, Biljana
AU  - Zečević, Mira
AU  - Malenović, Anđelija
AU  - Protić, Ana
PY  - 2022
UR  - https://farfar.pharmacy.bg.ac.rs/handle/123456789/4995
AB  - Most commonly used analytical technique for determination of active pharmaceutical ingredients and their impurities in quality control throughout all phases of drug research, development and manufacture is definitely reversed-phase high performance liquid chromatography (RP-HPLC). However, pharmaceutical industry professionals are often faced with various challenges in RP mode, which cannot be resolved with common variations in the composition of the mobile phase. These challenges often occur when analyzing compounds that contain basic ionizable groups, possess large differences in polarities and require consumption of high amounts of toxic organic solvents. Among available strategies for addressing the aforementioned issues, the most convenient one includes RP-HPLC mobile phase modifications by an addition of the proper chemical compounds. In that respect, RP-HPLC method can be easily adapted to the needs of the analysis without time-consuming and expensive equipment procurement. In this review the chaotropic chromatography, micellar liquid chromatography, and cyclodextrin modified RP-HPLC systems are presented and discussed in details. Special attention is devoted to the theoretical background, the possibility of retention modeling and applications in various fields of pharmacy, as well as their prospective in further research.
PB  - Elsevier B.V.
T2  - Journal of Chromatography Open
T1  - Modified aqueous mobile phases: A way to improve retention behavior of active pharmaceutical compounds and their impurities in liquid chromatography
VL  - 2
DO  - 10.1016/j.jcoa.2021.100023
ER  - 

@article{
author = "Đajić, Nevena and Krmar, Jovana and Rmandić, Milena and Rašević, Marija and Otašević, Biljana and Zečević, Mira and Malenović, Anđelija and Protić, Ana",
year = "2022",
abstract = "Most commonly used analytical technique for determination of active pharmaceutical ingredients and their impurities in quality control throughout all phases of drug research, development and manufacture is definitely reversed-phase high performance liquid chromatography (RP-HPLC). However, pharmaceutical industry professionals are often faced with various challenges in RP mode, which cannot be resolved with common variations in the composition of the mobile phase. These challenges often occur when analyzing compounds that contain basic ionizable groups, possess large differences in polarities and require consumption of high amounts of toxic organic solvents. Among available strategies for addressing the aforementioned issues, the most convenient one includes RP-HPLC mobile phase modifications by an addition of the proper chemical compounds. In that respect, RP-HPLC method can be easily adapted to the needs of the analysis without time-consuming and expensive equipment procurement. In this review the chaotropic chromatography, micellar liquid chromatography, and cyclodextrin modified RP-HPLC systems are presented and discussed in details. Special attention is devoted to the theoretical background, the possibility of retention modeling and applications in various fields of pharmacy, as well as their prospective in further research.",
publisher = "Elsevier B.V.",
journal = "Journal of Chromatography Open",
title = "Modified aqueous mobile phases: A way to improve retention behavior of active pharmaceutical compounds and their impurities in liquid chromatography",
volume = "2",
doi = "10.1016/j.jcoa.2021.100023"
}

Đajić, N., Krmar, J., Rmandić, M., Rašević, M., Otašević, B., Zečević, M., Malenović, A.,& Protić, A.. (2022). Modified aqueous mobile phases: A way to improve retention behavior of active pharmaceutical compounds and their impurities in liquid chromatography. in Journal of Chromatography Open
Elsevier B.V.., 2.
https://doi.org/10.1016/j.jcoa.2021.100023

Đajić N, Krmar J, Rmandić M, Rašević M, Otašević B, Zečević M, Malenović A, Protić A. Modified aqueous mobile phases: A way to improve retention behavior of active pharmaceutical compounds and their impurities in liquid chromatography. in Journal of Chromatography Open. 2022;2.
doi:10.1016/j.jcoa.2021.100023 .

Đajić, Nevena, Krmar, Jovana, Rmandić, Milena, Rašević, Marija, Otašević, Biljana, Zečević, Mira, Malenović, Anđelija, Protić, Ana, "Modified aqueous mobile phases: A way to improve retention behavior of active pharmaceutical compounds and their impurities in liquid chromatography" in Journal of Chromatography Open, 2 (2022),
https://doi.org/10.1016/j.jcoa.2021.100023 . .

TY  - CONF
AU  - Protić, Ana
AU  - Milenković, Milan
AU  - Rašević, Marija
AU  - Otašević, Biljana
AU  - Zečević, Mira
AU  - Krmar, Jovana
AU  - Malenović, Anđelija
PY  - 2022
UR  - https://farfar.pharmacy.bg.ac.rs/handle/123456789/4446
AB  - Nowadays, one of the ultimate requirements in drug analysis is rapid method
development, prompt chromatographic run time and long lasting method’s lifecycle. Special
attention should be paid to development of gradient elution (U)HPLC methods that are
commonly related to troubleshooting during the inter-laboratory method transfer. The dwell
volume difference between (U)HPLC systems is the main reason for this issue (1). The aim of
this study was to propose new gradient method development methodology with integrated
dwell volumes values in the optimization process. This is especially useful approach for
industry where is frequently known which (U)HPLC instruments will be applied for drug
analysis. Proposed methodology was tested on the model mixture which encompassed
dabigatran etexilate mesylate and its nine impurities. Experimental design methodology and
three different (U)HPLC instruments with significant dwell volume differences were utilized.
Statistically significant variables were selected with Plackett-Burman design, and along with
dwell volume values were included in D-optimal design. The separation criteria s between
adjacent peaks was selected as output for method optimization. Indirect modelling together
with Monte Carlo simulations enabled selection of optimal and robust chromatographic
conditions joint for all instruments. The optimal conditions included 24% (v/v) of initial
amount of acetonitrile, 54% (v/v) of the final amount of acetonitrile, 15 min of gradient
elution run time and pH value equal to 4.9. The proposed method utility was proved since it
was successfully validated and met all validation criteria (2).
AB  - U današnje vreme, jedan od važnih zahteva prilikom razvoja hromatografskih metoda
jeste da se razvoj uradi brzo, da vreme trajanja hromatografske analize bude kratko i da se
obezbedi dug životni ciklus metode. Posebnu pažnju treba obratiti na (U)HPLC metode sa
gradijentnim eluiranjem kod kojih se često javljaju problemi prilikom međulaboratorijskog
transfera. Glavni razlog za zahtevan transfer gradijentnih (U)HPLC metoda je razlika u
vrednostima dwell volume između (U)HPLC uređaja (1). Cilj ovog rada je bio da predloži
novu metodologiju razvoja gradijentnih (U)HPLC metoda, pri čemu će vrednosti dwell
volumes biti integrisane sa drugim ulaznim promenljivama u fazi optimizacije. Ovo bi bio
posebno koristan pristup u industriji gde se obično zna na kojim će sve (U)HPLC
instrumentima metoda biti korišćena. Metodologija je testirana na smeši dabigatran eteksilat
mezilata i njegovih devet nečistoća. Za razvoj metode korišćen je eksperimentalni dizajn i tri
različita (U)HPLC instrumenta sa značajnim razlikama u dwell volume vrednostima.
Statistički značajne ulazne promenljive su dobijene primenom Plackett‐Burman dizajna, i
zajedno sa vrednostima dwell volume su bile uključene u D‐opimal dizajn. Kriterijum
razdvajanja s između susednih parova pikova odabran je kao odgovor u odnosu na koji je
metoda optimizovana. Indirektno modelovanje zajedno sa Monte Karlo simulacijom
omoguć ilo je izbor optimalnih i robusnih hromatografskih uslova zajedničkih za sva tri
instrumenta. Oni su uključivali 24% (v/v) početnog udela acetonitrila, 54% (v/v) finalnog
udela acetonitrila, vreme trajanja gradijenta od 15 minuta i pH vrednost 4,9. Primenljivost
predložene metodologije je dokazana, jer je dobijena metoda uspešno validirana na sva tri
(U)HPLC instrumenta (2).
PB  - Savez farmaceutskih udruženja Srbije (SFUS)
C3  - Arhiv za farmaciju
T1  - Novel Gradient Elution (U)HPLC Method Development that Enables Successful Method Transfer
T1  - Novi pristup u razvoju (U)HPLC gradijentnih metoda koji bi omogućio uspešan transfer
VL  - 72
IS  - 4 suplement
SP  - S55
EP  - S56
UR  - https://hdl.handle.net/21.15107/rcub_farfar_4446
ER  - 

@conference{
author = "Protić, Ana and Milenković, Milan and Rašević, Marija and Otašević, Biljana and Zečević, Mira and Krmar, Jovana and Malenović, Anđelija",
year = "2022",
abstract = "Nowadays, one of the ultimate requirements in drug analysis is rapid method
development, prompt chromatographic run time and long lasting method’s lifecycle. Special
attention should be paid to development of gradient elution (U)HPLC methods that are
commonly related to troubleshooting during the inter-laboratory method transfer. The dwell
volume difference between (U)HPLC systems is the main reason for this issue (1). The aim of
this study was to propose new gradient method development methodology with integrated
dwell volumes values in the optimization process. This is especially useful approach for
industry where is frequently known which (U)HPLC instruments will be applied for drug
analysis. Proposed methodology was tested on the model mixture which encompassed
dabigatran etexilate mesylate and its nine impurities. Experimental design methodology and
three different (U)HPLC instruments with significant dwell volume differences were utilized.
Statistically significant variables were selected with Plackett-Burman design, and along with
dwell volume values were included in D-optimal design. The separation criteria s between
adjacent peaks was selected as output for method optimization. Indirect modelling together
with Monte Carlo simulations enabled selection of optimal and robust chromatographic
conditions joint for all instruments. The optimal conditions included 24% (v/v) of initial
amount of acetonitrile, 54% (v/v) of the final amount of acetonitrile, 15 min of gradient
elution run time and pH value equal to 4.9. The proposed method utility was proved since it
was successfully validated and met all validation criteria (2)., U današnje vreme, jedan od važnih zahteva prilikom razvoja hromatografskih metoda
jeste da se razvoj uradi brzo, da vreme trajanja hromatografske analize bude kratko i da se
obezbedi dug životni ciklus metode. Posebnu pažnju treba obratiti na (U)HPLC metode sa
gradijentnim eluiranjem kod kojih se često javljaju problemi prilikom međulaboratorijskog
transfera. Glavni razlog za zahtevan transfer gradijentnih (U)HPLC metoda je razlika u
vrednostima dwell volume između (U)HPLC uređaja (1). Cilj ovog rada je bio da predloži
novu metodologiju razvoja gradijentnih (U)HPLC metoda, pri čemu će vrednosti dwell
volumes biti integrisane sa drugim ulaznim promenljivama u fazi optimizacije. Ovo bi bio
posebno koristan pristup u industriji gde se obično zna na kojim će sve (U)HPLC
instrumentima metoda biti korišćena. Metodologija je testirana na smeši dabigatran eteksilat
mezilata i njegovih devet nečistoća. Za razvoj metode korišćen je eksperimentalni dizajn i tri
različita (U)HPLC instrumenta sa značajnim razlikama u dwell volume vrednostima.
Statistički značajne ulazne promenljive su dobijene primenom Plackett‐Burman dizajna, i
zajedno sa vrednostima dwell volume su bile uključene u D‐opimal dizajn. Kriterijum
razdvajanja s između susednih parova pikova odabran je kao odgovor u odnosu na koji je
metoda optimizovana. Indirektno modelovanje zajedno sa Monte Karlo simulacijom
omoguć ilo je izbor optimalnih i robusnih hromatografskih uslova zajedničkih za sva tri
instrumenta. Oni su uključivali 24% (v/v) početnog udela acetonitrila, 54% (v/v) finalnog
udela acetonitrila, vreme trajanja gradijenta od 15 minuta i pH vrednost 4,9. Primenljivost
predložene metodologije je dokazana, jer je dobijena metoda uspešno validirana na sva tri
(U)HPLC instrumenta (2).",
publisher = "Savez farmaceutskih udruženja Srbije (SFUS)",
journal = "Arhiv za farmaciju",
title = "Novel Gradient Elution (U)HPLC Method Development that Enables Successful Method Transfer, Novi pristup u razvoju (U)HPLC gradijentnih metoda koji bi omogućio uspešan transfer",
volume = "72",
number = "4 suplement",
pages = "S55-S56",
url = "https://hdl.handle.net/21.15107/rcub_farfar_4446"
}

Protić, A., Milenković, M., Rašević, M., Otašević, B., Zečević, M., Krmar, J.,& Malenović, A.. (2022). Novel Gradient Elution (U)HPLC Method Development that Enables Successful Method Transfer. in Arhiv za farmaciju
Savez farmaceutskih udruženja Srbije (SFUS)., 72(4 suplement), S55-S56.
https://hdl.handle.net/21.15107/rcub_farfar_4446

Protić A, Milenković M, Rašević M, Otašević B, Zečević M, Krmar J, Malenović A. Novel Gradient Elution (U)HPLC Method Development that Enables Successful Method Transfer. in Arhiv za farmaciju. 2022;72(4 suplement):S55-S56.
https://hdl.handle.net/21.15107/rcub_farfar_4446 .

Protić, Ana, Milenković, Milan, Rašević, Marija, Otašević, Biljana, Zečević, Mira, Krmar, Jovana, Malenović, Anđelija, "Novel Gradient Elution (U)HPLC Method Development that Enables Successful Method Transfer" in Arhiv za farmaciju, 72, no. 4 suplement (2022):S55-S56,
https://hdl.handle.net/21.15107/rcub_farfar_4446 .

TY  - 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 .

TY  - JOUR
AU  - Mitrović, Marija
AU  - Protić, Ana
AU  - Malenović, Anđelija
AU  - Otašević, Biljana
AU  - Zečević, Mira
PY  - 2020
UR  - https://farfar.pharmacy.bg.ac.rs/handle/123456789/3483
AB  - Official method in Ph. Eur. for evaluation of timolol enantiomeric purity is normal-phase high perfor-mance liquid chromatography (NP-HPLC) method. Compared to other HPLC modes, NP is depicted asquite expensive with high consumption of organic solvents which leads to chronic exposure of analyststo toxic and carcinogenic effects. In order to overcome above-mentioned drawbacks, the aim of thisstudy was to develop new method with better eco-friendly features. This was enabled by using proteintype Chiral Stationary Phase (CSP) in reversed-phase mode that required up to 10 % (v/v) of organicsolvent. Therefore, an enantioselective HPLC method was developed and validated for quantification of(S)-timolol and its chiral impurity, (R)-isomer. Optimized separation conditions on ovomucoid columnwere set using Analytical Quality by Design (AQbD) approach in method development. Optimizationstep was performed following the Box-Behnken experimental plan and the influence of three criticalmethod parameters (CMPs) towards enantioseparation of the above-mentioned peak pair was exam-ined. CMPs included variation of acetonitrile content in the mobile phase (5–10 %, v/v), pH value of theaqueous phase (6.0–7.0) and ammonium chloride concentration in the aqueous part of the mobile phase(10−30 mmol L−1). The most relevant critical method attributes (CMAs) in this case were the separa-tion criterion between studied critical pair and retention factor of the second eluting peak, (S)-timolol.Qualitative Design Space (DS) was defined by Monte Carlo simulations providing adequate assurance ofmethod’s qualitative robustness ( = 95 %). The selected working point situated in the middle of the DSwas characterized by following combination of CMPs: acetonitrile content in the mobile phase 7 % (v/v),pH value of the aqueous phase 6.8 and concentration of ammonium chloride in aqueous phase 14 mmolL–1. In the next step, the quantitative robustness was tested by Plackett-Burman experimental design. Thevalidation studies confirmed adequacy of the proposed method for its intended purpose. Finally, Ana-lytical Eco-Scale metric tool was applied to confirm that developed method represents excellent greenanalytical method compared to the official one.
PB  - Elsevier
T2  - Journal of Pharmaceutical and Biomedical Analysis
T1  - Analytical quality by design development of an ecologically acceptable enantioselective HPLC method for timolol maleate enantiomeric purity testing on ovomucoid chiral stationary phase
VL  - 180
DO  - 10.1016/j.jpba.2019.113034
ER  - 

@article{
author = "Mitrović, Marija and Protić, Ana and Malenović, Anđelija and Otašević, Biljana and Zečević, Mira",
year = "2020",
abstract = "Official method in Ph. Eur. for evaluation of timolol enantiomeric purity is normal-phase high perfor-mance liquid chromatography (NP-HPLC) method. Compared to other HPLC modes, NP is depicted asquite expensive with high consumption of organic solvents which leads to chronic exposure of analyststo toxic and carcinogenic effects. In order to overcome above-mentioned drawbacks, the aim of thisstudy was to develop new method with better eco-friendly features. This was enabled by using proteintype Chiral Stationary Phase (CSP) in reversed-phase mode that required up to 10 % (v/v) of organicsolvent. Therefore, an enantioselective HPLC method was developed and validated for quantification of(S)-timolol and its chiral impurity, (R)-isomer. Optimized separation conditions on ovomucoid columnwere set using Analytical Quality by Design (AQbD) approach in method development. Optimizationstep was performed following the Box-Behnken experimental plan and the influence of three criticalmethod parameters (CMPs) towards enantioseparation of the above-mentioned peak pair was exam-ined. CMPs included variation of acetonitrile content in the mobile phase (5–10 %, v/v), pH value of theaqueous phase (6.0–7.0) and ammonium chloride concentration in the aqueous part of the mobile phase(10−30 mmol L−1). The most relevant critical method attributes (CMAs) in this case were the separa-tion criterion between studied critical pair and retention factor of the second eluting peak, (S)-timolol.Qualitative Design Space (DS) was defined by Monte Carlo simulations providing adequate assurance ofmethod’s qualitative robustness ( = 95 %). The selected working point situated in the middle of the DSwas characterized by following combination of CMPs: acetonitrile content in the mobile phase 7 % (v/v),pH value of the aqueous phase 6.8 and concentration of ammonium chloride in aqueous phase 14 mmolL–1. In the next step, the quantitative robustness was tested by Plackett-Burman experimental design. Thevalidation studies confirmed adequacy of the proposed method for its intended purpose. Finally, Ana-lytical Eco-Scale metric tool was applied to confirm that developed method represents excellent greenanalytical method compared to the official one.",
publisher = "Elsevier",
journal = "Journal of Pharmaceutical and Biomedical Analysis",
title = "Analytical quality by design development of an ecologically acceptable enantioselective HPLC method for timolol maleate enantiomeric purity testing on ovomucoid chiral stationary phase",
volume = "180",
doi = "10.1016/j.jpba.2019.113034"
}

Mitrović, M., Protić, A., Malenović, A., Otašević, B.,& Zečević, M.. (2020). Analytical quality by design development of an ecologically acceptable enantioselective HPLC method for timolol maleate enantiomeric purity testing on ovomucoid chiral stationary phase. in Journal of Pharmaceutical and Biomedical Analysis
Elsevier., 180.
https://doi.org/10.1016/j.jpba.2019.113034

Mitrović M, Protić A, Malenović A, Otašević B, Zečević M. Analytical quality by design development of an ecologically acceptable enantioselective HPLC method for timolol maleate enantiomeric purity testing on ovomucoid chiral stationary phase. in Journal of Pharmaceutical and Biomedical Analysis. 2020;180.
doi:10.1016/j.jpba.2019.113034 .

Mitrović, Marija, Protić, Ana, Malenović, Anđelija, Otašević, Biljana, Zečević, Mira, "Analytical quality by design development of an ecologically acceptable enantioselective HPLC method for timolol maleate enantiomeric purity testing on ovomucoid chiral stationary phase" in Journal of Pharmaceutical and Biomedical Analysis, 180 (2020),
https://doi.org/10.1016/j.jpba.2019.113034 . .

TY  - CONF
AU  - Mitrović, Marija
AU  - Malenović, Anđelija
AU  - Otašević, Biljana
AU  - Protić, Ana
AU  - Zečević, Mira
PY  - 2019
UR  - https://farfar.pharmacy.bg.ac.rs/handle/123456789/4706
PB  - University of Milano-Bicocca
C3  - 48th International Symposium on High-Performance Liquid Phase Separations and Related Techniques HPLC 2019, June 16-20, 2019, Milan, Italy.
T1  - Development and validation of a HPLC method for the enantiomeric purity testing of timolol maleate on ovomucoid chiral stationary phase
UR  - https://hdl.handle.net/21.15107/rcub_farfar_4706
ER  - 

@conference{
author = "Mitrović, Marija and Malenović, Anđelija and Otašević, Biljana and Protić, Ana and Zečević, Mira",
year = "2019",
publisher = "University of Milano-Bicocca",
journal = "48th International Symposium on High-Performance Liquid Phase Separations and Related Techniques HPLC 2019, June 16-20, 2019, Milan, Italy.",
title = "Development and validation of a HPLC method for the enantiomeric purity testing of timolol maleate on ovomucoid chiral stationary phase",
url = "https://hdl.handle.net/21.15107/rcub_farfar_4706"
}

Mitrović, M., Malenović, A., Otašević, B., Protić, A.,& Zečević, M.. (2019). Development and validation of a HPLC method for the enantiomeric purity testing of timolol maleate on ovomucoid chiral stationary phase. in 48th International Symposium on High-Performance Liquid Phase Separations and Related Techniques HPLC 2019, June 16-20, 2019, Milan, Italy.
University of Milano-Bicocca..
https://hdl.handle.net/21.15107/rcub_farfar_4706

Mitrović M, Malenović A, Otašević B, Protić A, Zečević M. Development and validation of a HPLC method for the enantiomeric purity testing of timolol maleate on ovomucoid chiral stationary phase. in 48th International Symposium on High-Performance Liquid Phase Separations and Related Techniques HPLC 2019, June 16-20, 2019, Milan, Italy.. 2019;.
https://hdl.handle.net/21.15107/rcub_farfar_4706 .

Mitrović, Marija, Malenović, Anđelija, Otašević, Biljana, Protić, Ana, Zečević, Mira, "Development and validation of a HPLC method for the enantiomeric purity testing of timolol maleate on ovomucoid chiral stationary phase" in 48th International Symposium on High-Performance Liquid Phase Separations and Related Techniques HPLC 2019, June 16-20, 2019, Milan, Italy. (2019),
https://hdl.handle.net/21.15107/rcub_farfar_4706 .