Original scientific paper

https://doi.org/10.2478/acph-2020-0046

Miniaturized shake-flask HPLC method for determination of distribution coefficient of drugs used in inflammatory bowel diseases

EDVIN BRUSAČ ; University of Zagreb Faculty of Pharmacy and Biochemistry 10 000 Zagreb, Croatia
MARIO-LIVIO JELIČIĆ ; University of Zagreb Faculty of Pharmacy and Biochemistry 10 000 Zagreb, Croatia
DANIELA AMIDŽIĆ KLARIĆ orcid.org/0000-0002-1695-6950 ; University of Zagreb Faculty of Pharmacy and Biochemistry 10 000 Zagreb, Croatia
ANA MORNAR orcid.org/0000-0002-3944-697X ; University of Zagreb Faculty of Pharmacy and Biochemistry 10 000 Zagreb, Croatia

Abstract

A new method for determination of distribution coefficient of drugs azathioprine, 6-mercaptopurine and 6-thioguanine and nutrient folic acid used in the treatment of inflammatory bowel disease based on a miniaturized shake-flask and HPLC/DAD was developed. Special attention was made to the most commonly reported problems in the measurement of distribution coefficients using a shake-flask method such as mixing technique, speed and time, the temperature of experiment, type of buffer and its pH as well as n-octanol/buffer phase ratio. The concentration of compounds in the buffer is determined by HPLC directly from shake flasks or conventional 2-mL vials. The developed method was fully validated according to ICH guidelines. Furthermore, experimental data were successfully compared with lipophilicity and human intestinal absorption calculated by the use of four different theoretical approaches. The method shows potential for high-throughput measurements of a large number of compounds.

Keywords

distribution coefficient logarithm; shake-flask; HPLC; in silico; inflammatory bowel disease

Hrčak ID:

220976

URI

https://hrcak.srce.hr/220976

Publication date:

31.12.2019.

Visits: 1.873 *