`Stirred not shaken!' Comparing agitation methods for permeability studies using a novel type of 96-well sandwich-plates

Jonas Borregaard Eriksen, Ann-Christin Jacobsen, Katrine Tækker Christensen, Annette Bauer-Brandl, Martin Brandl

Research output: Contribution to journalJournal articleResearchpeer-review

Abstract

In order to achieve a high sample throughput, permeation experiments are often carried out using 96-well sandwich plates. Even though agitation is regarded as important, permeation studies in 96-well format are often carried out without agitation since orbital shaking, the most common agitation method for 96-well plates, has been reported to create difficulties (e.g., well-to-well cross-talk), and high cost and low availability limits the use of other agitation techniques (e.g., magnetic stirring). This study investigates how orbital shaking and magnetic stirring affect the apparent permeability of model compounds with different water-solubilities (methylene blue, carbamazepine, and albendazole) using a novel 96-well sandwich plate comprising a cellulose-hydrate membrane (PermeaPlain® plate). Orbital shaking was found less efficient than magnetic stirring in terms of homogeneously distributing a small volume of dye within the donor compartment. Furthermore, in terms of achieving maximum trans-barrier flux, magnetic stirring was found a more effective agitation method than orbital shaking. Obviously, with orbital shaking the medium in the bottom compartment of the sandwich plates never was mixed in-phase. The impact of insufficient mixing on permeation was found strongest with the most lipophilic compound, which correlates with literature reports that the contribution of the unstirred water layer towards the overall resistance of the barrier is most expressed in case of lipophilic drugs. Finally, it was tested how different liquid volumes in the bottom compartment of the plates affect the well-to-well cross-talk during permeation experiments under orbital shaking. This study revealed that 250-300 µL should be used in the bottom compartment of the sandwich plates to reduce well-to-well cross-talk when using orbital shaking for agitation.

Original languageEnglish
JournalJournal of Pharmaceutical Sciences
ISSN0022-3549
DOIs
Publication statusE-pub ahead of print - 5. Jun 2021

Bibliographical note

Copyright © 2021. Published by Elsevier Inc.

Fingerprint

Dive into the research topics of '`Stirred not shaken!' Comparing agitation methods for permeability studies using a novel type of 96-well sandwich-plates'. Together they form a unique fingerprint.

Cite this