Title:Box-bhenken Design Combined with 3D Surface Methodology for Optimization of an Eco-friendly HPLC Method to Determine Venetoclax in Human Plasma, and its Bioanalytical Method Validation According to ICH M10 Guideline
Volume: 22
Issue: 7
Author(s): Saniye Özcan*, Abeer Elriş, Mazlum Akif Altun, Serkan Levent and Nafiz Öncü Can
Affiliation:
- Department of Analytical Chemistry, Faculty of Pharmacy, Anadolu University, 26470, Eskisehir, Türkiye
- Central
Analysis Laboratory (MERLAB), Faculty of Pharmacy, Anadolu University, 26470, Eskisehir, Türkiye
Keywords:
Box-behnken design, green chemistry, HPLC, ICH M10, venetoclax, 3D surface methodology, reversed phase, C8.
Abstract:
Aims: Venetoclax is a selective inhibitor of the prosurvival protein BCL-2 approved by
the Food Drug Administration in 2016, restoring the apoptic ability of malignant cells. In this study,
a fast, highly accurate and precise HPLC method was developed for the analysis of Venetoclax in
human plasma.
Methods: The optimization of the method was investigated according to Box-Bhenken Design combined
with 3D surface methodology. The chromatographic separation was performed in gradient
mode with an Ascentis Express C8 column (2.7 μm, 4.6 mm × 10 cm). Agomelatine was used as an
internal standard to increase accuracy. The method was completely validated according to ICH
guideline M10 bioanalytical method validation. Additionally, the greenness of the method was scaled
with NEMI, Analytical Ecoscale, AGREE, and GAPI greenness metrics.
Results: The method was linear in the range of 1.67-12.50 μg/mL with a calculated R2 of 0.99; LOD
and LOQ were 0.34 μg/mL and 1.02 μg/mL, respectively. The recovery was in the range of 102.6%
to 99.08%, and with an RSD% of less than 1.00%. The analytical eco scale and AGREE score of the
current method were 85 and 0.55, respectively.
Conclusion: The approach that was developed herein exhibits green, rapidity, high levels of accuracy
and precision, cost-effectiveness, and ease of use in the context of clinical and pharmacokinetic
investigations.
