Chemometrically Assisted Optimization of Pregabalin Fluorescent Derivatization Reaction with a Novel Xanthone Analogue and Validation of the Method for the Determination of Pregabalin in Bulk via a Plate Reader
Quantitation of chromophore-free analytes is always a challenge. To this purpose, derivatization of the analyte constitutes a common strategy, leading to a product with a strong signal. In the current study, a novel xanthone analogue was utilized for the first time for the derivatization of pregabal...
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| Format: | Article |
| Language: | English |
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MDPI AG
2022-03-01
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| Series: | Molecules |
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| Online Access: | https://www.mdpi.com/1420-3049/27/6/1954 |
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| author | Nikolaos Kritikos Aikaterini Iliou Amalia D. Kalampaliki Evangelos Gikas Ioannis K. Kostakis Benoît Y. Michel Yannis Dotsikas |
| author_facet | Nikolaos Kritikos Aikaterini Iliou Amalia D. Kalampaliki Evangelos Gikas Ioannis K. Kostakis Benoît Y. Michel Yannis Dotsikas |
| author_sort | Nikolaos Kritikos |
| collection | DOAJ |
| description | Quantitation of chromophore-free analytes is always a challenge. To this purpose, derivatization of the analyte constitutes a common strategy, leading to a product with a strong signal. In the current study, a novel xanthone analogue was utilized for the first time for the derivatization of pregabalin, a model analyte with a primary amine moiety that lacks a chromophore. The fact that only the xanthene-based derivative, formed after the derivatization reaction fluoresces, enables avoiding its chromatographic separation from the reagent and thus reducing the analysis time of a series of samples in 1–2 min via a plate reader. The reaction conditions were optimized via a central composite design (CCD), with fluorescence signal as the measure of the yield. The following factors that affect the derivatization reaction were chosen: (a) temperature, (b) reaction time, and (c) triethylamine solution volume used to drive the reaction to completion. After the identification of the optimal conditions, the method was validated according to ICH guidelines, using a fluorescence plate reader for signal measurement (λ<sub>ex</sub> = 540, λ<sub>em</sub> = 615 nm). Finally, the newly developed high-throughput method was applied to the determination of drug content in pregabalin bulk. |
| first_indexed | 2024-03-09T13:09:24Z |
| format | Article |
| id | doaj.art-28fecfde92294dbd8a688a669aa4982a |
| institution | Directory Open Access Journal |
| issn | 1420-3049 |
| language | English |
| last_indexed | 2024-03-09T13:09:24Z |
| publishDate | 2022-03-01 |
| publisher | MDPI AG |
| record_format | Article |
| series | Molecules |
| spelling | doaj.art-28fecfde92294dbd8a688a669aa4982a2023-11-30T21:43:45ZengMDPI AGMolecules1420-30492022-03-01276195410.3390/molecules27061954Chemometrically Assisted Optimization of Pregabalin Fluorescent Derivatization Reaction with a Novel Xanthone Analogue and Validation of the Method for the Determination of Pregabalin in Bulk via a Plate ReaderNikolaos Kritikos0Aikaterini Iliou1Amalia D. Kalampaliki2Evangelos Gikas3Ioannis K. Kostakis4Benoît Y. Michel5Yannis Dotsikas6Laboratory of Pharmaceutical Analysis, Department of Pharmacy, National and Kapodistrian University of Athens, Panepistimioupoli Zografou, GR-157 71 Athens, GreeceLaboratory of Pharmaceutical Analysis, Department of Pharmacy, National and Kapodistrian University of Athens, Panepistimioupoli Zografou, GR-157 71 Athens, GreeceDivision of Pharmaceutical Chemistry, Department of Pharmacy, National and Kapodistrian University of Athens, Panepistimioupoli Zografou, GR-157 71 Athens, GreeceLaboratory of Analytical Chemistry, Department of Chemistry, National and Kapodistrian University of Athens, Panepistimioupoli Zografou, GR-157 71 Athens, GreeceDivision of Pharmaceutical Chemistry, Department of Pharmacy, National and Kapodistrian University of Athens, Panepistimioupoli Zografou, GR-157 71 Athens, GreeceInstitut de Chimie de Nice, CNRS, UMR 7272, Université Côte d’Azur, Parc Valrose, CEDEX 2, 06108 Nice, FranceLaboratory of Pharmaceutical Analysis, Department of Pharmacy, National and Kapodistrian University of Athens, Panepistimioupoli Zografou, GR-157 71 Athens, GreeceQuantitation of chromophore-free analytes is always a challenge. To this purpose, derivatization of the analyte constitutes a common strategy, leading to a product with a strong signal. In the current study, a novel xanthone analogue was utilized for the first time for the derivatization of pregabalin, a model analyte with a primary amine moiety that lacks a chromophore. The fact that only the xanthene-based derivative, formed after the derivatization reaction fluoresces, enables avoiding its chromatographic separation from the reagent and thus reducing the analysis time of a series of samples in 1–2 min via a plate reader. The reaction conditions were optimized via a central composite design (CCD), with fluorescence signal as the measure of the yield. The following factors that affect the derivatization reaction were chosen: (a) temperature, (b) reaction time, and (c) triethylamine solution volume used to drive the reaction to completion. After the identification of the optimal conditions, the method was validated according to ICH guidelines, using a fluorescence plate reader for signal measurement (λ<sub>ex</sub> = 540, λ<sub>em</sub> = 615 nm). Finally, the newly developed high-throughput method was applied to the determination of drug content in pregabalin bulk.https://www.mdpi.com/1420-3049/27/6/1954fluorescencexanthonederivatization reactionplate readerpregabalinexperimental design |
| spellingShingle | Nikolaos Kritikos Aikaterini Iliou Amalia D. Kalampaliki Evangelos Gikas Ioannis K. Kostakis Benoît Y. Michel Yannis Dotsikas Chemometrically Assisted Optimization of Pregabalin Fluorescent Derivatization Reaction with a Novel Xanthone Analogue and Validation of the Method for the Determination of Pregabalin in Bulk via a Plate Reader Molecules fluorescence xanthone derivatization reaction plate reader pregabalin experimental design |
| title | Chemometrically Assisted Optimization of Pregabalin Fluorescent Derivatization Reaction with a Novel Xanthone Analogue and Validation of the Method for the Determination of Pregabalin in Bulk via a Plate Reader |
| title_full | Chemometrically Assisted Optimization of Pregabalin Fluorescent Derivatization Reaction with a Novel Xanthone Analogue and Validation of the Method for the Determination of Pregabalin in Bulk via a Plate Reader |
| title_fullStr | Chemometrically Assisted Optimization of Pregabalin Fluorescent Derivatization Reaction with a Novel Xanthone Analogue and Validation of the Method for the Determination of Pregabalin in Bulk via a Plate Reader |
| title_full_unstemmed | Chemometrically Assisted Optimization of Pregabalin Fluorescent Derivatization Reaction with a Novel Xanthone Analogue and Validation of the Method for the Determination of Pregabalin in Bulk via a Plate Reader |
| title_short | Chemometrically Assisted Optimization of Pregabalin Fluorescent Derivatization Reaction with a Novel Xanthone Analogue and Validation of the Method for the Determination of Pregabalin in Bulk via a Plate Reader |
| title_sort | chemometrically assisted optimization of pregabalin fluorescent derivatization reaction with a novel xanthone analogue and validation of the method for the determination of pregabalin in bulk via a plate reader |
| topic | fluorescence xanthone derivatization reaction plate reader pregabalin experimental design |
| url | https://www.mdpi.com/1420-3049/27/6/1954 |
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