Abstract
A new analytical method using high-performance liquid chromatography coupled with photo diode array detection was developed and validated for the quantification of Diclofenac (DIC) from in vitro skin permeation samples. Analysis was performed using a Phenomenex C18 column (150 x 4.6mm, 5µm) with 10mM ammonium acetate: Acetonitrile (62:38% v/v) as the mobile phase in isocratic mode and eluents were monitored at 276nm. DIC was eluted at 3.1min and showed a good linearity in the concentration range of 0.2-3µg/mL with a correlation coefficient >0.999. The validation parameters, such as specificity, linearity, accuracy and limit of detection, limit of quantification, precision, robustness fulfilled the regulatory requirements. The developed HPLC method was successfully used for the analysis of DIC in samples obtained from transdermal diffusate samples.
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