Journal of Bioequivalence & Bioavailability

A rapid HPLC method for metoprolol quantification: Application for ex vivo intestinal studies using Franz cells

6^th World Congress on Bioavailability & Bioequivalence: BA/BE Studies Summit

August 17-19, 2015 Chicago, USA

Andre B Dezani, Thaisa M Dezani, Mônica M C Silva and Cristina H R Serra

Posters-Accepted Abstracts: J Bioequiv Availab

Abstract:

An important factor for the good bioavailability of a drug is the complete absorption and studies involving this prediction have
been grown in recent years. Among several studies, ex vivo methods using vertical diffusion chamber (Franz cells) experiments
have gained highlight for permeation studies. This method employs an intestinal segment and it can be used for biowaiver purposes.
This work aims to show a chromatographic method developed and validated for quantification of metoprolol contained in samples
obtained from ex vivo assays using Franz cells. A simple, precise, specific and accurate RP-HPLC (reversed-phase high performance
liquid chromatography) method was developed and validated using ICH guideline considering parameters as selectivity/specificity,
linearity, precision, accuracy, limit of detection and limit of quantification. The chromatographic conditions were achieved with:
Gemini C18 column (250 mm x 4.6 mm x 5 μm), fluorescence detection set at 225 nm for excitation and 310 nm for emission, oven
set at 35°C. The mobile phase consisted of a mixture of phosphate buffer 20 mM and acetonitrile (80:20%) pH 4.5, flow of 1 mL/min
and injection volume of 25 μL. The assessed parameters were in accordance with the ICH recommendations. The method presented
linearity in a range of 10-1000 ng/mL (r²=0.9993). Precision and accuracy are also adequate with intraday and interday variation no
higher than 5%. During development and validation, no interferences were noticed in chromatograms, showing the selectivity and
specificity of the method. The chromatographic method was developed and validated for metoprolol quantification in samples from
Franz cells assay and its applicability in daily lab routine is possible for permeation studies with several kinds of biological tissues,
including intestine. Also, this method can be useful for development of other chromatographic methods in similar studies.

Biography :

Andre B Dezani has completed his Master’s degree in 2010 and since then, he is developing his PhD thesis related to permeability studies using different methods as ex vivo
and in vitro models. His research field also includes solubility, biopharmaceutical classification systems, dissolution studies and ADME prediction. The studies are conducted
in Faculty of Pharmaceutical Sciences of University of Sao Paulo, Brazil.