Chemometrically Assisted RP-HPLC Method Development for Efficient Separation of Ivabradine and its Eleven Impurities
Аутори
Tomić, JovanaIvković, Branka
Oljačić, Slavica
Nikolić, Katarina
Maljurić, Nevena
Protić, Ana
Agbaba, Danica
Чланак у часопису (Објављена верзија)
Метаподаци
Приказ свих података о документуАпстракт
The aim of this study was to develop a novel reversed-phase high-performance liquid chromatography (RP-HPLC)
method for efficient separation of ivabradine and its 11 impurities. Similar polarity of impurities in the sample mixture
made method optimization challenging and accomplishable only when different chemometric tools, such as
principal component analysis (PCA), Box–Behnken design (BBD), and desirability function as a multicriteria approach,
were employed. The presence of 3 positional isomers (impurities III, V, and VI), keto–enol tautomerism of
impurity VII, and diastereoisomers of impurity X made separation of this complex mixture even more challenging.
Chromatographic retention parameters obtained with the mobile phase consisting of 30 mM phosphate buffer and
acetonitrile (80:20, v/v) on four different RP-HPLC columns at varying pH values (3.0, 4.0, and 5.0) were subjected
to the PCA analysis to select the column with the most appropriate selectivity. Then the column te...mperature,
pH of the aqueous component of mobile phase, phosphate buffer molarity and the organic solvent content in the
mobile phase were estimated employing BBD. Valid and reliable mathematical models towards resolution of twelve
critical peak pairs were obtained. After determination of the desirability making criteria for all responses, desirability
functions were established and used in optimization. The proposed optimal chromatographic conditions included
the Zorbax Eclipse Plus C18 chromatographic column (100 × 4.6 mm, 3.5 μm), the column temperature of 34 °C,
the mobile phase flow rate of 1.6 mL min−1 and the UV detection at 220 nm. The mobile phase consisted of the
28 mM phosphate buffer at pH 6.0 and acetonitrile (85:15, v/v). Separation of one pair of positional isomers was
not achieved, so methanol was added to the organic part of mobile phase in small increments with the optimal ratio
of methanol to acetonitrile 59:41, v/v. The overall organic component of the mobile phase also increased to 18%,
accelerating the chromatographic analysis.
Кључне речи:
Ivabradine / principal component analysis / Box-Behnken design / isocratic elution / RP-HPLC / diastereoisomersИзвор:
Acta Chromatographica, 2020, 32, 1, 53-63Издавач:
- Akademiai Kiado Zrt.
Финансирање / пројекти:
- Синтеза, квантитативни однос између структуре и дејства, физичко-хемијска карактеризација и анализа фармаколошки активних супстанци (RS-MESTD-Basic Research (BR or ON)-172033)
DOI: 10.1556/1326.2019.00659
ISSN: 1233-2356
WoS: 000520838000010
Scopus: 2-s2.0-85086134969
Институција/група
PharmacyTY - JOUR AU - Tomić, Jovana AU - Ivković, Branka AU - Oljačić, Slavica AU - Nikolić, Katarina AU - Maljurić, Nevena AU - Protić, Ana AU - Agbaba, Danica PY - 2020 UR - https://farfar.pharmacy.bg.ac.rs/handle/123456789/4903 AB - The aim of this study was to develop a novel reversed-phase high-performance liquid chromatography (RP-HPLC) method for efficient separation of ivabradine and its 11 impurities. Similar polarity of impurities in the sample mixture made method optimization challenging and accomplishable only when different chemometric tools, such as principal component analysis (PCA), Box–Behnken design (BBD), and desirability function as a multicriteria approach, were employed. The presence of 3 positional isomers (impurities III, V, and VI), keto–enol tautomerism of impurity VII, and diastereoisomers of impurity X made separation of this complex mixture even more challenging. Chromatographic retention parameters obtained with the mobile phase consisting of 30 mM phosphate buffer and acetonitrile (80:20, v/v) on four different RP-HPLC columns at varying pH values (3.0, 4.0, and 5.0) were subjected to the PCA analysis to select the column with the most appropriate selectivity. Then the column temperature, pH of the aqueous component of mobile phase, phosphate buffer molarity and the organic solvent content in the mobile phase were estimated employing BBD. Valid and reliable mathematical models towards resolution of twelve critical peak pairs were obtained. After determination of the desirability making criteria for all responses, desirability functions were established and used in optimization. The proposed optimal chromatographic conditions included the Zorbax Eclipse Plus C18 chromatographic column (100 × 4.6 mm, 3.5 μm), the column temperature of 34 °C, the mobile phase flow rate of 1.6 mL min−1 and the UV detection at 220 nm. The mobile phase consisted of the 28 mM phosphate buffer at pH 6.0 and acetonitrile (85:15, v/v). Separation of one pair of positional isomers was not achieved, so methanol was added to the organic part of mobile phase in small increments with the optimal ratio of methanol to acetonitrile 59:41, v/v. The overall organic component of the mobile phase also increased to 18%, accelerating the chromatographic analysis. PB - Akademiai Kiado Zrt. T2 - Acta Chromatographica T1 - Chemometrically Assisted RP-HPLC Method Development for Efficient Separation of Ivabradine and its Eleven Impurities VL - 32 IS - 1 SP - 53 EP - 63 DO - 10.1556/1326.2019.00659 ER -
@article{ author = "Tomić, Jovana and Ivković, Branka and Oljačić, Slavica and Nikolić, Katarina and Maljurić, Nevena and Protić, Ana and Agbaba, Danica", year = "2020", abstract = "The aim of this study was to develop a novel reversed-phase high-performance liquid chromatography (RP-HPLC) method for efficient separation of ivabradine and its 11 impurities. Similar polarity of impurities in the sample mixture made method optimization challenging and accomplishable only when different chemometric tools, such as principal component analysis (PCA), Box–Behnken design (BBD), and desirability function as a multicriteria approach, were employed. The presence of 3 positional isomers (impurities III, V, and VI), keto–enol tautomerism of impurity VII, and diastereoisomers of impurity X made separation of this complex mixture even more challenging. Chromatographic retention parameters obtained with the mobile phase consisting of 30 mM phosphate buffer and acetonitrile (80:20, v/v) on four different RP-HPLC columns at varying pH values (3.0, 4.0, and 5.0) were subjected to the PCA analysis to select the column with the most appropriate selectivity. Then the column temperature, pH of the aqueous component of mobile phase, phosphate buffer molarity and the organic solvent content in the mobile phase were estimated employing BBD. Valid and reliable mathematical models towards resolution of twelve critical peak pairs were obtained. After determination of the desirability making criteria for all responses, desirability functions were established and used in optimization. The proposed optimal chromatographic conditions included the Zorbax Eclipse Plus C18 chromatographic column (100 × 4.6 mm, 3.5 μm), the column temperature of 34 °C, the mobile phase flow rate of 1.6 mL min−1 and the UV detection at 220 nm. The mobile phase consisted of the 28 mM phosphate buffer at pH 6.0 and acetonitrile (85:15, v/v). Separation of one pair of positional isomers was not achieved, so methanol was added to the organic part of mobile phase in small increments with the optimal ratio of methanol to acetonitrile 59:41, v/v. The overall organic component of the mobile phase also increased to 18%, accelerating the chromatographic analysis.", publisher = "Akademiai Kiado Zrt.", journal = "Acta Chromatographica", title = "Chemometrically Assisted RP-HPLC Method Development for Efficient Separation of Ivabradine and its Eleven Impurities", volume = "32", number = "1", pages = "53-63", doi = "10.1556/1326.2019.00659" }
Tomić, J., Ivković, B., Oljačić, S., Nikolić, K., Maljurić, N., Protić, A.,& Agbaba, D.. (2020). Chemometrically Assisted RP-HPLC Method Development for Efficient Separation of Ivabradine and its Eleven Impurities. in Acta Chromatographica Akademiai Kiado Zrt.., 32(1), 53-63. https://doi.org/10.1556/1326.2019.00659
Tomić J, Ivković B, Oljačić S, Nikolić K, Maljurić N, Protić A, Agbaba D. Chemometrically Assisted RP-HPLC Method Development for Efficient Separation of Ivabradine and its Eleven Impurities. in Acta Chromatographica. 2020;32(1):53-63. doi:10.1556/1326.2019.00659 .
Tomić, Jovana, Ivković, Branka, Oljačić, Slavica, Nikolić, Katarina, Maljurić, Nevena, Protić, Ana, Agbaba, Danica, "Chemometrically Assisted RP-HPLC Method Development for Efficient Separation of Ivabradine and its Eleven Impurities" in Acta Chromatographica, 32, no. 1 (2020):53-63, https://doi.org/10.1556/1326.2019.00659 . .