Quantitative estimation and validation of isotretinoin in Pharmaceutical dosage forms by RP-HPLC

Abstract

Isotretinoin is a topical keratolytic agent which is used in the treatment of skin diseases including acne vulgaris.
This paper deals with a simple, feasible and sensitive reverse-phase high-performance liquid chromatographic
method for the quantitative determination of isotretinoin in pharmaceutical capsule dosage form. The
chromatography was carried out by using HPLC system (Shimadzu LC2010HT) with UV- Visible dual
absorbance detector (PDA), C18, 25 cm X 4.6 mm, 5 µm. The mobile phase consisting of 0.3% glacial acetic
acid and water in the ratio of 85:15 and tetrahydrofuran and methanol used a diluent in the ratio of 20:80. The
detection was made at 355 nm and the mobile phase flowed at 1 ml/min. Validation parameters included system
suitability, specificity, linearity, accuracy precision (repeatability & reproducibility), robustness, ruggedness and
stability were determined according to the ICH guidelines. The retention time of isotretinoin was found to be
12.33 min. Hence, the method could be successfully applied for routine analysis of isotretinoin in
pharmaceutical dosage forms.