Stability indicating RP-HPLC method development and validation for quantification of impurities in gonadotropin-releasing hormone (Elagolix): Robustness study by quality by design

Biomed Chromatogr. 2024 Dec;38(12):e6036. doi: 10.1002/bmc.6036. Epub 2024 Nov 1.

Abstract

The purpose of this research was to establish and validate a reverse phase HPLC method for the determination of Elagolix impurities in pharmaceutical dosage form. Mobile phase A, consisting of 10 mM sodium dihydrogen phosphate (pH 6.0) and acetonitrile in a 95:5 v/v ratio, and mobile phase B, containing 85:10:5 v/v/v of acetonitrile, Milli-Q water, and methanol, were used to achieve the method's specificity in the analytical column Kromasil 100-C18 (250 mm × 4.6 mm, 5 μm). The gradient program includes (%B/Time [min]: 36/0, 36/10, 38/15, 85/55, 85/65, 36/67, and 36/75). The flow rate is 0.8 mL/min. The overall run duration is 75.0 min, the injection volume is 10.0 μL, and the detection is at 210 nm in UV. The samples were subjected to hydrolysis, oxidation, and heat conditions in order to facilitate their forced degradation. The procedure was validated and determined with the standards of ICH guidelines. From the LOQ to a concentration level of 200%, the linearity of the technique was ascertained. An accuracy range of LOQ to 150% was established for the method, and the average recovery was acceptable. Design of experiments, part of the quality by design idea, was used to prove the method's reliability.

Keywords: Elagolix; HPLC; ICH; analytical method validation; forced degradation; quality by design.

MeSH terms

  • Chromatography, High Pressure Liquid / methods
  • Chromatography, Reverse-Phase* / methods
  • Drug Contamination*
  • Drug Stability*
  • Gonadotropin-Releasing Hormone* / analogs & derivatives
  • Gonadotropin-Releasing Hormone* / analysis
  • Limit of Detection*
  • Linear Models
  • Reproducibility of Results

Substances

  • Gonadotropin-Releasing Hormone