Development and Validation of a High-Performance Thin-Layer Chromatography (HPTLC) Method for Accurate Quantification of Cyclosporine in Pharmaceutical Formulations

This study presents the development of a new HPTLC method for the quantification of cyclosporine in bulk drug substances and pharmaceutical formulations, adhering to ICH guidelines. The technique employs a mobile phase consisting of toluene, ethyl acetate, and acetic acid (3:7). Cyclosporine is detected using densitometric scanning at 202 nm, with separation performed on silica gel 60 F254 TLC plates (0.2 mm). The method demonstrates excellent linearity (R² > 0.999) over the concentration range of 50–150%. Accuracy studies show recoveries within 98–102%, while precision analysis reveals %RSD values below 2.0%. Specificity testing confirms that cyclosporine is well resolved from excipients and impurities, ensuring accurate quantification. The limit of detection (LOD) is determined to be 44.02 ng/band, and the limit of quantification (LOQ) is 124.3 ng/band, highlighting the method's high sensitivity. A robustness assessment indicates that variations in chamber saturation time and mobile phase composition have minimal impact on the method's performance. Due to its simplicity, low cost, and reliable performance, the developed HPTLC method is ideal for routine quality control of cyclosporine in pharmaceutical formulations. This work provides a foundation for using this method in stability testing, impurity identification, and the determination of therapeutic drug levels in body fluids.