Background The hidradenitis suppurativa (HS) is an inflammatory skin disease. Resorcinol is a phenol derivate, and in topical self-treatment decreases the size and pain of HS lesions. Topical 15% resorcinol is prepared as a pharmaceutical compound. In current literature, high performance liquid chromatography (HPLC) methods have been reported for determination of resorcinol, using different gradients of mobile phase mainly.
Purpose To assess the correct formulation, our objective is to develop an isocratic HPLC method for quality control.
Material and methods To develop and validate the method, an Agilent 1260 HPLC system was used. Performed at 25°C, the separation was carried out in reverse phase column (Agilent Zorbax Eclipse XDB-C18 4.6 mm x 250 mm, 5 µm i. d.) (USA) with an isocratic mode. The mobile phase was methanol:water 40%:60%, the flow rate was 1 mL/minute and the injection volume was 10 mcL. A diode array detector was used (=280 nm). All reagents were analytical grade and bought from Sigma-Aldrich (USA). The calibration line was generated by least squares linear regression. The method accuracy (acceptance criterion of 99%–101%) and precision performed in a within-day and between-day anaylisis (five consecutive days) was assessed by five replicates run (low-, medium- and high-level concentration) and <1% variability as acceptance criterion. Spectral analysis 2D and 3D was performed to assess specificity and selectivity of method.
Results There was an adequate separation under the chromatografic conditions presented. The retention time (mean ±SD) was 3.62±0.006 min. UV spectrum analysis demonstrated the specificity and selectivity of the method, since there were no co-eluates at the same retention time of resorcinol.
The calibration line was y=8.6584x+11.8571(‘Y’:area under the curve and ‘X’:experimental concentration). It was linear (R2=0.99993). All the standards passed the acceptance criteria. The mean recovery percentage was 99.994% in all concentration levels. The coefficient of variation within-day was 0.082%, 0.042% and 0.128% for low-, medium- and high-point respectively, and between-day was 0.016%, 0.199% and 0.221% for low-, medium- and high-point respectively.
Conclusion The presented method shows a simplified way to analyse resorcinol by HPLC using UV detector. The performance of the method has been proven in terms of linearity, accuracy, precision, selectivity and specificity by a validation assay.
References and/or Acknowledgements doi:10. 5740/jaoacint. 14–123
No conflict of interest