Analytical Method Development and Validation Using RP-HPLC for Cyclosporine (CsA) Formulated in ‘Liquid Crystalline Nanoparticles (LCN’S)’: Application in Determining Drug Loading(%DL), Drug Entrapment (%DEE) and Drug Release Profile

Abstract

Abstract Background Cyclosporine (CsA)is used as an antifungal, immunosuppressant administered orally. But it exhibit poor aqueous solubility due to the presence of lipophilic, cyclic endecapeptide in large fraction, which makes poor permeability across the membranes. So, liquid crystallinenano formulation could enhance the solubility properties. However, to date, there is no analytical method development or validation pertinent to cyclosporine (CsA) as liquid crystalline Nano formulations. Objective To develop and validate a simple and quick reverse phase, high-performance liquid chromatographic method for determination of cyclosporine (CsA) using UV detector. Employ the method to determine drug loading (%DL), drug entrapment efficiency (%DEE) and to quantify drug release samples. Methods The mobile phase included acetonitrile and 10Mm ammonium acetate as a buffering media. In the ratio 90:10 (ACN: Ammonium Acetate, pH 4) using Shim-pack GIST C18 (4.6 x 100 mm, 5 µm). Column with flow rate of 0.8ml/min, with column temperature (50 ºC), the eluent detected at 215nm with injection volume 20µl. Results The determination of cyclosporine (CsA) was accurate and precise based on the data pertinent to system adaptability, accuracy, linearity, precision, specificity, and robustness. This approach was validated in accordance with ICH guidelines. The LOD & LOQ found to be 0.12 µg/ml and 0.15µg/ml. According to the approach, the linearity was in the range of 1–64 µg/ml. It has been claimed that cyclosporine (CsA) recovery rates range from 98 to 100%. Because there was no invasion from excipients or mobile phase, the strategy was proven to be accurate. Cyclosporine (CsA) has a run time of around 10 minutes. The calibration curve was accurate. Cyclosporine (CsA) elution time was 4.56 minutes. The accuracy of the system was 98.2% respectively, and the %RSD was less than 2%. The % entrapment efficiency of CsA -LCN was found to be 83.13%. Whereas the % DL for CsA -LCN was found to be 5.90 ± 0.31. Drug release profile showed a sustained release pattern for CsA -LCN in comparison to pure drug Conclusion RP-HPLC method is a sensitive, precise, accurate method for analytical method development and validation specifically for (CsA) in liquid crystalline nanoparticles.