Development and validation of a reverse phase liquid chromatography method for the simultaneous quantification of eserine and pralidoxime chloride in drugs-in-adhesive matrix type transdermal patches.

BACKGROUND

In the present study, a simple, precise, specific, fast, accurate and reliable reverse phase high performance liquid chromatographic (RP-HPLC) method has been developed and validated for the simultaneous determination and quantification of eserine and pralidoxime chloride in drugs-in-adhesive matrix type transdermal patches.

METHODS

The chromatographic separation was achieved by C18 column, using a mobile phase consisting of acetonitrile: 10 mM potassium dihydrogen phosphate, 10 mM heptane-1-sulfonic acid sodium salt monohydrate in water (30:70, v/v) adjusted at pH 3.0 with ortho-phosphoric acid. Flow rate was 1.0 mL/min and UV detection at 238 nm. The method was validated according to the International Conference on Harmonization (ICH) guidelines.

RESULTS

The calibration curves were linear over the different concentration ranges of 0.5-10 μg/ml for eserine and 5-25 μg/mL for 2PAM. Relative standard deviation for precision was less than 2.0%. Limit of detection values of eserine and 2-PAM were 0.018 µg/mL and 0.008 µg/mL, respectively. The limit of quantification of eserine and 2-PAM were 0.055 µg/mL and 0.026 µg/mL, respectively.

CONCLUSION

The developed method was applied for the routine analysis of these 2 drugs in drugs-in-adhesive matrix type transdermal patches in order to evaluate the drug content of different formulations. It could be also used with reliability for the determination of the drug in other pharmaceutical dosage forms.