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Analytical Quality by Design: Achieving Robustness of an LC-CAD Method for the Analysis of Non-Volatile Fatty Acids

Authors :
Rasmus Walther
Jovana Krmar
Adrian Leistner
Bojana Svrkota
Biljana Otašević
Andjelija Malenović
Ulrike Holzgrabe
Ana Protić
Source :
Pharmaceuticals; Volume 16; Issue 4; Pages: 478, Pharmaceuticals
Publication Year :
2023
Publisher :
Multidisciplinary Digital Publishing Institute, 2023.

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 C18 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% (v/v) within 14.2 min.

Details

Language :
English
ISSN :
14248247
Database :
OpenAIRE
Journal :
Pharmaceuticals; Volume 16; Issue 4; Pages: 478
Accession number :
edsair.doi.dedup.....d3b51b00fea3f6788af8479e181e0d60
Full Text :
https://doi.org/10.3390/ph16040478