Integrating analytical quality by design into bioanalytical method development: an HPLC-FLD method for quantification of alectinib in biological matrix
Abstract This study presents the development and validation of a fluorescence-based high-performance liquid chromatography (HPLC) method for the quantification of alectinib in rat plasma, with a focus on the application of Analytical Quality by Design (AQbD) to bioanalytical method development. Unli...
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| Format: | Article |
| Language: | English |
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BMC
2025-08-01
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| Series: | BMC Chemistry |
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| Online Access: | https://doi.org/10.1186/s13065-025-01613-z |
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| author | Eun Ji Lee Nahyun Koo Min Ju Kim Kyeong-Ryoon Lee Yoon-Jee Chae |
| author_facet | Eun Ji Lee Nahyun Koo Min Ju Kim Kyeong-Ryoon Lee Yoon-Jee Chae |
| author_sort | Eun Ji Lee |
| collection | DOAJ |
| description | Abstract This study presents the development and validation of a fluorescence-based high-performance liquid chromatography (HPLC) method for the quantification of alectinib in rat plasma, with a focus on the application of Analytical Quality by Design (AQbD) to bioanalytical method development. Unlike conventional QbD applications, which primarily address synthetic formulations or instrumental settings, this study systematically applied AQbD principles to the complex environment of biological matrices. Critical method parameters, including the organic phase ratio, buffer concentration, and flow rate, were identified through Failure Mode and Effects Analysis, and optimized using a Box–Behnken design. The final method exhibited excellent linearity (R² >0.99) over a concentration range of 5–1,000 ng/mL, with a lower limit of quantification of 5 ng/mL. It also showed high accuracy (95.6–102%), precision (relative standard deviation < 11%), and consistent recovery (98.3–105%), with minimal matrix effects. Alectinib stability was confirmed under various handling conditions. This method was successfully applied in a pharmacokinetic study after intravenous and oral administration of alectinib in rats. These results highlight the value of AQbD in addressing specific challenges of bioanalysis and demonstrate its utility in establishing a sensitive, robust, and regulatory-compliant method suitable for pharmacokinetic applications. |
| format | Article |
| id | doaj-art-ddb15e9bd7134faf9d5f0367c35ed514 |
| institution | Kabale University |
| issn | 2661-801X |
| language | English |
| publishDate | 2025-08-01 |
| publisher | BMC |
| record_format | Article |
| series | BMC Chemistry |
| spelling | doaj-art-ddb15e9bd7134faf9d5f0367c35ed5142025-08-24T11:06:01ZengBMCBMC Chemistry2661-801X2025-08-0119111410.1186/s13065-025-01613-zIntegrating analytical quality by design into bioanalytical method development: an HPLC-FLD method for quantification of alectinib in biological matrixEun Ji Lee0Nahyun Koo1Min Ju Kim2Kyeong-Ryoon Lee3Yoon-Jee Chae4College of Pharmacy, Woosuk UniversityCollege of Pharmacy, Woosuk UniversityLaboratory Animal Resource Center, Korea Research Institute of Bioscience and BiotechnologyLaboratory Animal Resource Center, Korea Research Institute of Bioscience and BiotechnologyCollege of Pharmacy, Woosuk UniversityAbstract This study presents the development and validation of a fluorescence-based high-performance liquid chromatography (HPLC) method for the quantification of alectinib in rat plasma, with a focus on the application of Analytical Quality by Design (AQbD) to bioanalytical method development. Unlike conventional QbD applications, which primarily address synthetic formulations or instrumental settings, this study systematically applied AQbD principles to the complex environment of biological matrices. Critical method parameters, including the organic phase ratio, buffer concentration, and flow rate, were identified through Failure Mode and Effects Analysis, and optimized using a Box–Behnken design. The final method exhibited excellent linearity (R² >0.99) over a concentration range of 5–1,000 ng/mL, with a lower limit of quantification of 5 ng/mL. It also showed high accuracy (95.6–102%), precision (relative standard deviation < 11%), and consistent recovery (98.3–105%), with minimal matrix effects. Alectinib stability was confirmed under various handling conditions. This method was successfully applied in a pharmacokinetic study after intravenous and oral administration of alectinib in rats. These results highlight the value of AQbD in addressing specific challenges of bioanalysis and demonstrate its utility in establishing a sensitive, robust, and regulatory-compliant method suitable for pharmacokinetic applications.https://doi.org/10.1186/s13065-025-01613-zAlectinibAnalytical quality by designTaguchi methodBox–Behnken designMethod development and validation |
| spellingShingle | Eun Ji Lee Nahyun Koo Min Ju Kim Kyeong-Ryoon Lee Yoon-Jee Chae Integrating analytical quality by design into bioanalytical method development: an HPLC-FLD method for quantification of alectinib in biological matrix BMC Chemistry Alectinib Analytical quality by design Taguchi method Box–Behnken design Method development and validation |
| title | Integrating analytical quality by design into bioanalytical method development: an HPLC-FLD method for quantification of alectinib in biological matrix |
| title_full | Integrating analytical quality by design into bioanalytical method development: an HPLC-FLD method for quantification of alectinib in biological matrix |
| title_fullStr | Integrating analytical quality by design into bioanalytical method development: an HPLC-FLD method for quantification of alectinib in biological matrix |
| title_full_unstemmed | Integrating analytical quality by design into bioanalytical method development: an HPLC-FLD method for quantification of alectinib in biological matrix |
| title_short | Integrating analytical quality by design into bioanalytical method development: an HPLC-FLD method for quantification of alectinib in biological matrix |
| title_sort | integrating analytical quality by design into bioanalytical method development an hplc fld method for quantification of alectinib in biological matrix |
| topic | Alectinib Analytical quality by design Taguchi method Box–Behnken design Method development and validation |
| url | https://doi.org/10.1186/s13065-025-01613-z |
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