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Modification of gradient HPLC method for determination of small molecules' affinity to human serum albumin under column safety conditions: Robustness and chemometrics study
Abstract
In the early stages of drug discovery, beyond the biological activity screening, determining the physicochemical properties that affect the distribution of molecules in the human body is an essential step. Plasma protein binding (PPB) is one of the most important investigated endpoints. Nevertheless, the methodology for measuring %PPB is significantly less popular and standardized than other physicochemical properties, like lipophilicity. Here, we proposed how to modify protocols presented by Valko into column safety conditions and evaluated their robustness using fractional factorial design. For robustness testing, four factors were selected: column temperature, mobile phase flow rate, maximum isopropanol concentration in the mobile phase, and buffer pH. Elaborate methods have been applied for the analysis of HSA affinity for three groups of antibiotic-oriented substances that vary in chemical structure: fluoroquinolones, sulfonamides, and tetrazole derivatives. Furthermore, based on the reversed-phase chromatography the workflow of pilot studies was proposed to select molecules that have high affinity to HSA and can not be eluted from the HSA column using the concentration of organic modifier recommended by the column manufacturer
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Authors (10)
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Mateusz Woziński
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Katarzyna Greber
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Monika Pastewska
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Piotr Kolasiński
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Beata Żołnowska
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Jarosław Sławiński
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Daniel Szulczyk
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Wiesław Sawicki
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Krzesimir Ciura
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Details
- Category:
- Articles
- Type:
- artykuły w czasopismach
- Published in:
-
JOURNAL OF PHARMACEUTICAL AND BIOMEDICAL ANALYSIS
no. 239,
ISSN: 0731-7085 - Language:
- English
- Publication year:
- 2024
- Bibliographic description:
- Woziński M., Greber K., Pastewska M., Kolasiński P., Hewelt-Belka W., Żołnowska B., Sławiński J., Szulczyk D., Sawicki W., Ciura K.: Modification of gradient HPLC method for determination of small molecules' affinity to human serum albumin under column safety conditions: Robustness and chemometrics study// JOURNAL OF PHARMACEUTICAL AND BIOMEDICAL ANALYSIS -Vol. 239, (2024), s.115916-
- DOI:
- Digital Object Identifier (open in new tab) 10.1016/j.jpba.2023.115916
- Sources of funding:
-
- Free publication
- Verified by:
- Gdańsk University of Technology
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