The aim of this work is to quantify the Ibuprofen sodium salt in samples derived from Franz cells transdermal delivery experiments.
An analytical method for a determination of Ibuprofen sodium salt and Diclofenac sodium salt (used as internal standard) has been developed, assuring rapidity, accuracy, precision, selectivity and a minimal treatment of the sample.
With this method we have been able to analized not only the sample obtained from the recipient cell, but also the skin and the formulation used in the experiment.
To this aim a methods that would allow the easy removal of the drug from the tissue has been developped.
In this way, it was easy to verify the real quantification of the drug in all phases of the experiment and quantify its total delivery.
In order to perform High Performance Liquid Cromatography (HPLC) analysis, human skin used in experiment has been dissolved first and then deproteinized.
The effectiveness of deproteinization performed through methods such as precipitation with organic solvent, cryo-treatment, Amicon filter tubes, HybridSpe cartridges and Folch method has been verified through the use of the NanoOrange kit.
For the HPLC method, the working conditions were as follows: RP18 column EC 150/4,6 NUCLEODUR® 100-5 C18 ec, Macherey-Nagel GmbH & Co. KG protected with a guard column (EC 4/3 NUCLEODUR®, 100-5 C18 ec, 100-5 18 ec, Macherey-Nagel GmbH & Co. KG) and a pre-column (cut-off: 0,2 μm, Supelco, n. cat. 55215-U); mobile phase phosphate buffer 25 mM - Acetonitrile (73:27 v/v) (pH 7) adjusted with hydrochloric acid; flow 1.0 mL/min.; injection volume of 20 μL; and UV detection was carried out at 265 nm, and experiment were performed in a room where temperature was constantly kept at 25°C.
With this method, the detection of a drug concentration in a range between 0.03 - 7.69 μg/mL has been allowed.
Therefore, the proposed method can be successfully used for drug analysis in samples derived from transdermal delivery experiments
