Background: The aim of this study was to develop HPLC methods for individual determination
of 20 active pharmaceutical ingredients (amoxicillin sodium, antipyrine, atenolol, caffeine, carbamazepine,
cimetidine, enalapril, furosemide, hydrochlorothiazide, ibuprofen, ketoprofen, metoprolol tartrate,
methyldopa, naproxen sodium, pindolol, piroxicam, propranolol HCl, ranitidine, theophylline, and
verapamil HCl) to be used for determination of their intestinal permeabilities across Ussing-Chamber.
Method: Two different stationary phases (Waters X-Bridge C18- 150 x 4.6 mm, 5 µm; and ACE 5 C18- 150
x 4.6 mm, 5 µm) were used for the separation of the compounds. Three different aqueous phases (20 mM
phosphate buffers pH 3.0, pH 6.0 and water) and two different organic phases (methanol and acetonitrile)
were used to prepare the mobile phases. Total analysis time was shorter than 7 minutes for all applications.
Result: The developed methods were validated according to the ICH guideline and found to be linear,
sensitive, selective, precise and accurate. The developed methods could be applied for analyses of these
compounds not only for Ussing-Chamber studies but also for other permeability studies.