Abstract
Isotretinoin content was accurately estimated using UV and RP-HPLC methods. A simple, sensitive double-beam spectrophotometric technique was determined for the assessment of Isotretinoin content in pure form as well as its formulations. Standard solution of Isotretinoin showed its highest absorbance at 354 nm. Beer’s Law was found to be linear between 1–10 µg/ml, with a correlation coefficient (r²) of 0.9983, a slope of 0.0791, and an intercept of 0.0119. The method was validated and exhibited good linearity, precision and accuracy for quantitative estimation of Isotretinoin in tablet formulations. The method was optimized using a mobile phase of Acetonitrile and methanol (30:70) at a flow rate of 1.2 ml/min to accomplish good resolution. The present method had a good correlation coefficient (r2) of 0.9992 and linear in the concentration range 1–64 μg/ml. Study protocol for content estimation was rapid. Retention time of the drug was found to be 2.592 min, with high accuracy (% recovery = 97.7, 97.7, 98.3), precision, and sensitivity. The literature reveals that no systematic scientific evaluation has been carried out for isotretinoin using both the UV and RP-HPLC methods. The process was validated according to the ICH guidelines, ensuring that the technique is reliable and reproducible one.
Keywords: 13-Cis-Retinoic Acid, ICH-Guidelines, Isotretinoin, Method Development, RP-HPLC, Validation.