Ateş M.Kaynak M.S.Şahin S.2024-08-042024-08-0420151300-0608https://hdl.handle.net/11616/91151Intestinal perfusion (SPIP) technique is one of the most commonly used techniques to determine the intestinal permeability of a drug. In perfusion studies, metoprolol (as tartrate) and phenol red are widely used as a refer- ence compounds to evaluate the permeability coefficient of the compound of interest (acyclovir in this study). The aim of our study was to develop and validate a reversed-phase liquid chromatographic method for the simultan- eous determination of acyclovir, metoprolol and phenol red for use in intes- tinal perfusion studies. The analysis was performed on a C18 column (4.6 mm x 250 mm, 5 ?m) using a mobile phase consisting of methanol:0.0125 M potassium dihydrogen phosphate buffer (55:45, v/v; pH 7.0). Method was validated according to the FDA guidelines for selectivity, sensitivity, linearity, precision, accuracy, stability. All calibration curves were linear (r2> 0.999). Lower limit of quantitation was 0.04 ?g/mL for acyclovir, 0.02 ?g/mL for metoprolol, 0.01 ?g/mL for phenol red. Detection limit was 0.01 ?g/mL for acyclovir 0.002 ?g/mL for metoprolol, 0.003 ?g/mL for phenol red. Precision and accuracy results of the method fulfilled the required limits. This newly developed and validated method can be readily used on a routine basis for the standardization of in situ intestinal permeability experiments. © 2015, Hacettepe University, Faculty of Pharmacy. All rights reserved.eninfo:eu-repo/semantics/closedAccessAcyclovirHPLCIn situ intestinal perfusionMetoprololPhenol redArticle3521461612-s2.0-84971307677Q4