Analytical Quality by Design: Achieving Robustness of an LC-CAD Method for the Analysis of Non-Volatile Fatty Acids
An alternative to the time-consuming and error-prone pharmacopoeial gas chromatography method for the analysis of fatty acids (FAs) is urgently needed. The objective was therefore to propose a robust liquid chromatography method with charged aerosol detection for the analysis of polysorbate 80 (PS80...
| Main Authors: | , , , , , , , |
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| Format: | Article |
| Language: | English |
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MDPI AG
2023-03-01
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| Series: | Pharmaceuticals |
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| Online Access: | https://www.mdpi.com/1424-8247/16/4/478 |
| _version_ | 1827744048455090176 |
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| author | Rasmus Walther Jovana Krmar Adrian Leistner Bojana Svrkota Biljana Otašević Andjelija Malenović Ulrike Holzgrabe Ana Protić |
| author_facet | Rasmus Walther Jovana Krmar Adrian Leistner Bojana Svrkota Biljana Otašević Andjelija Malenović Ulrike Holzgrabe Ana Protić |
| author_sort | Rasmus Walther |
| collection | DOAJ |
| description | An alternative to the time-consuming and error-prone pharmacopoeial gas chromatography method for the analysis of fatty acids (FAs) is urgently needed. The objective was therefore to propose a robust liquid chromatography method with charged aerosol detection for the analysis of polysorbate 80 (PS80) and magnesium stearate. FAs with different numbers of carbon atoms in the chain necessitated the use of a gradient method with a Hypersil Gold C<sub>18</sub> column and acetonitrile as organic modifier. The risk-based Analytical Quality by Design approach was applied to define the Method Operable Design Region (MODR). Formic acid concentration, initial and final percentages of acetonitrile, gradient elution time, column temperature, and mobile phase flow rate were identified as critical method parameters (CMPs). The initial and final percentages of acetonitrile were fixed while the remaining CMPs were fine-tuned using response surface methodology. Critical method attributes included the baseline separation of adjacent peaks (α-linolenic and myristic acid, and oleic and petroselinic acid) and the retention factor of the last compound eluted, stearic acid. The MODR was calculated by Monte Carlo simulations with a probability equal or greater than 90%. Finally, the column temperature was set at 33 °C, the flow rate was 0.575 mL/min, and acetonitrile linearly increased from 70 to 80% (<i>v</i>/<i>v</i>) within 14.2 min. |
| first_indexed | 2024-03-11T04:39:28Z |
| format | Article |
| id | doaj.art-8ef32e416f0b4464afdde9d101cbad72 |
| institution | Directory Open Access Journal |
| issn | 1424-8247 |
| language | English |
| last_indexed | 2024-03-11T04:39:28Z |
| publishDate | 2023-03-01 |
| publisher | MDPI AG |
| record_format | Article |
| series | Pharmaceuticals |
| spelling | doaj.art-8ef32e416f0b4464afdde9d101cbad722023-11-17T20:49:30ZengMDPI AGPharmaceuticals1424-82472023-03-0116447810.3390/ph16040478Analytical Quality by Design: Achieving Robustness of an LC-CAD Method for the Analysis of Non-Volatile Fatty AcidsRasmus Walther0Jovana Krmar1Adrian Leistner2Bojana Svrkota3Biljana Otašević4Andjelija Malenović5Ulrike Holzgrabe6Ana Protić7Institute for Pharmacy and Food Chemistry, University of Würzburg, Am Hubland, 97074 Würzburg, GermanyDepartment of Drug Analysis, Faculty of Pharmacy, University of Belgrade, Vojvode Stepe 450, 11 221 Belgrade, SerbiaInstitute for Pharmacy and Food Chemistry, University of Würzburg, Am Hubland, 97074 Würzburg, GermanyDepartment of Drug Analysis, Faculty of Pharmacy, University of Belgrade, Vojvode Stepe 450, 11 221 Belgrade, SerbiaDepartment of Drug Analysis, Faculty of Pharmacy, University of Belgrade, Vojvode Stepe 450, 11 221 Belgrade, SerbiaDepartment of Drug Analysis, Faculty of Pharmacy, University of Belgrade, Vojvode Stepe 450, 11 221 Belgrade, SerbiaInstitute for Pharmacy and Food Chemistry, University of Würzburg, Am Hubland, 97074 Würzburg, GermanyDepartment of Drug Analysis, Faculty of Pharmacy, University of Belgrade, Vojvode Stepe 450, 11 221 Belgrade, SerbiaAn alternative to the time-consuming and error-prone pharmacopoeial gas chromatography method for the analysis of fatty acids (FAs) is urgently needed. The objective was therefore to propose a robust liquid chromatography method with charged aerosol detection for the analysis of polysorbate 80 (PS80) and magnesium stearate. FAs with different numbers of carbon atoms in the chain necessitated the use of a gradient method with a Hypersil Gold C<sub>18</sub> column and acetonitrile as organic modifier. The risk-based Analytical Quality by Design approach was applied to define the Method Operable Design Region (MODR). Formic acid concentration, initial and final percentages of acetonitrile, gradient elution time, column temperature, and mobile phase flow rate were identified as critical method parameters (CMPs). The initial and final percentages of acetonitrile were fixed while the remaining CMPs were fine-tuned using response surface methodology. Critical method attributes included the baseline separation of adjacent peaks (α-linolenic and myristic acid, and oleic and petroselinic acid) and the retention factor of the last compound eluted, stearic acid. The MODR was calculated by Monte Carlo simulations with a probability equal or greater than 90%. Finally, the column temperature was set at 33 °C, the flow rate was 0.575 mL/min, and acetonitrile linearly increased from 70 to 80% (<i>v</i>/<i>v</i>) within 14.2 min.https://www.mdpi.com/1424-8247/16/4/478Analytical Quality by Designfatty acidscharged aerosol detectorpolysorbate 80magnesium stearate |
| spellingShingle | Rasmus Walther Jovana Krmar Adrian Leistner Bojana Svrkota Biljana Otašević Andjelija Malenović Ulrike Holzgrabe Ana Protić Analytical Quality by Design: Achieving Robustness of an LC-CAD Method for the Analysis of Non-Volatile Fatty Acids Pharmaceuticals Analytical Quality by Design fatty acids charged aerosol detector polysorbate 80 magnesium stearate |
| title | Analytical Quality by Design: Achieving Robustness of an LC-CAD Method for the Analysis of Non-Volatile Fatty Acids |
| title_full | Analytical Quality by Design: Achieving Robustness of an LC-CAD Method for the Analysis of Non-Volatile Fatty Acids |
| title_fullStr | Analytical Quality by Design: Achieving Robustness of an LC-CAD Method for the Analysis of Non-Volatile Fatty Acids |
| title_full_unstemmed | Analytical Quality by Design: Achieving Robustness of an LC-CAD Method for the Analysis of Non-Volatile Fatty Acids |
| title_short | Analytical Quality by Design: Achieving Robustness of an LC-CAD Method for the Analysis of Non-Volatile Fatty Acids |
| title_sort | analytical quality by design achieving robustness of an lc cad method for the analysis of non volatile fatty acids |
| topic | Analytical Quality by Design fatty acids charged aerosol detector polysorbate 80 magnesium stearate |
| url | https://www.mdpi.com/1424-8247/16/4/478 |
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