In Silico Plasma Protein Binding Studies of Selected Group of Drugs Using TLC and HPLC Retention Data

Plasma protein binding is an important determinant of the pharmacokinetic properties of chemical compounds in living organisms. The aim of the present study was to determine the index of protein binding affinity based on chromatographic experiments. The question is which chromatographic environment will best mimic the drug–protein binding conditions. Retention data from normal phase thin-layer liquid chromatography (NP TLC), reversed phase (RP) TLC and HPLC chromatography experiments with 129 active pharmaceutical ingredients (APIs) were collected. The stationary phase of the TLC plates was modified with protein and the HPLC column was filled with immobilized human serum albumin. In both chromatographic methods, the mobile phase was based on a buffer with a pH of 7.4 to mimic physiological conditions. Chemometric analyses were performed to compare multiple linear regression models (MLRs) with retention data, using protein binding values as the dependent variable. In the course of the analysis, APIs were divided into acidic, basic and neutral groups, and separate models were created for each group. The MLR models had a coefficient of determination between 0.73 and 0.91, with the highest values from NP TLC data.

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Ultrafiltration Method for Plasma Protein Binding Studies and Its Limitations

Processes ◽

10.3390/pr9020382 ◽

2021 ◽

Vol 9 (2) ◽

pp. 382

Author(s):

Camelia-Maria Toma ◽

Silvia Imre ◽

Camil-Eugen Vari ◽

Daniela-Lucia Muntean ◽

Amelia Tero-Vescan

Keyword(s):

Protein Binding ◽

Plasma Protein Binding ◽

Plasma Protein ◽

Specific Binding ◽

Critical Role ◽

Volume Ratio ◽

Binding Studies ◽

Experimental Conditions ◽

Key Aspects

Plasma protein binding plays a critical role in drug therapy, being a key part in the characterization of any compound. Among other methods, this process is largely studied by ultrafiltration based on its advantages. However, the method also has some limitations that could negatively influence the experimental results. The aim of this study was to underline key aspects regarding the limitations of the ultrafiltration method, and the potential ways to overcome them. The main limitations are given by the non-specific binding of the substances, the effect of the volume ratio obtained, and the need of a rigorous control of the experimental conditions, especially pH and temperature. This review presents a variety of methods that can hypothetically reduce the limitations, and concludes that ultrafiltration remains a reliable method for the study of protein binding. However, the methodology of the study should be carefully chosen.

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Correlation between reversed-phase high-performance liquid chromatography and plasma protein binding

Journal of Chromatography B Biomedical Sciences and Applications ◽

10.1016/0378-4347(87)80381-x ◽

1987 ◽

Vol 421 ◽

pp. 83-90 ◽

Cited By ~ 12

Author(s):

J. Ganansia ◽

G. Bianchetti ◽

J.P. Thénot

Keyword(s):

High Performance Liquid Chromatography ◽

Liquid Chromatography ◽

Protein Binding ◽

Plasma Protein Binding ◽

Plasma Protein ◽

High Performance ◽

Reversed Phase

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Pharmacokinetics, Tissue Distribution, Plasma Protein Binding Studies of 10-Dehydroxyl-12-Demethoxy-Conophylline, a Novel Anti-Tumor Candidate, in Rats

Molecules ◽

10.3390/molecules24020283 ◽

2019 ◽

Vol 24 (2) ◽

pp. 283

Author(s):

Chengjun Jiang ◽

Jie Li ◽

Xianghai Cai ◽

Nini Li ◽

Yan Guo ◽

...

Keyword(s):

Protein Binding ◽

Tissue Distribution ◽

Plasma Protein Binding ◽

Plasma Protein ◽

Equilibrium Dialysis ◽

Free Fraction ◽

Ultra Performance Liquid Chromatography ◽

Sprague Dawley ◽

Binding Studies ◽

High Protein Binding

10-Dehydroxyl-12-demethoxy-conophylline is a natural anticancer candidate. The motivation of this study was to explore the pharmacokinetic profiles, tissue distribution, and plasma protein binding of 10-dehydroxyl-12-demethoxy-conophylline in Sprague Dawley rats. A rapid, sensitive, and specific ultra-performance liquid chromatography (UPLC) system with a fluorescence (FLR) detection method was developed for the determination of 10-dehydroxyl-12-demethoxy-conophylline in different rat biological samples. After intravenous (i.v.) dosing of 10-dehydroxyl-12-demethoxy-conophylline at different levels (4, 8, and 12 mg/kg), the half-life t1/2α of intravenous administration was about 7 min and the t1/2β was about 68 min. The AUC0→∞ increased in a dose-proportional manner from 68.478 μg/L·min for 4 mg/kg to 305.616 mg/L·min for 12 mg/kg. After intragastrical (i.g.) dosing of 20 mg/kg, plasma levels of 10-dehydroxyl-12-demethoxy-conophylline peaked at about 90 min. 10-dehydroxyl-12-demethoxy-conophyllinea absolute oral bioavailability was only 15.79%. The pharmacokinetics process of the drug was fit to a two-room model. Following a single i.v. dose (8 mg/kg), 10-dehydroxyl-12-demethoxy-conophylline was detected in all examined tissues with the highest in kidney, liver, and lung. Equilibrium dialysis was used to evaluate plasma protein binding of 10-dehydroxyl-12-demethoxy-conophylline at three concentrations (1.00, 2.50, and 5.00 µg/mL). Results indicated a very high protein binding degree (over 80%), reducing substantially the free fraction of the compound.

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