| Title: | Analytical Quality by Design: Achieving Robustness of an LC-CAD Method for the Analysis of Non-Volatile Fatty Acids |
| Author(s): | Walther R; Krmar J; Leistner A; Svrkota B; Otasevic B; Malenovic A; Holzgrabe U; Protic A; |
| Address: | "Institute for Pharmacy and Food Chemistry, University of Wurzburg, Am Hubland, 97074 Wurzburg, Germany. Department of Drug Analysis, Faculty of Pharmacy, University of Belgrade, Vojvode Stepe 450, 11 221 Belgrade, Serbia" |
| ISSN/ISBN: | 1424-8247 (Print) 1424-8247 (Electronic) 1424-8247 (Linking) |
| Abstract: | "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(18) 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 (alpha-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 degrees C, the flow rate was 0.575 mL/min, and acetonitrile linearly increased from 70 to 80% (v/v) within 14.2 min" |
| Keywords: | Analytical Quality by Design charged aerosol detector fatty acids magnesium stearate polysorbate 80; |
| Notes: | "PubMed-not-MEDLINEWalther, Rasmus Krmar, Jovana Leistner, Adrian Svrkota, Bojana Otasevic, Biljana Malenovic, Andjelija Holzgrabe, Ulrike Protic, Ana eng Grant Agreement with University of Belgrade-Faculty of Pharmacy No: 451-03-47/2023-01/200161/Ministry of Education and Science of Germany and the Ministry of Education, Science and Technological Development, Republic of Serbia/ Switzerland 2023/04/28 Pharmaceuticals (Basel). 2023 Mar 23; 16(4):478. doi: 10.3390/ph16040478" |
